Methods of treating cancer with anti-MICA / B antibodies

Abstract

Provided herein are methods of treating cancer with an antibody that specifically binds to MICA / B, or an antigen-binding fragment thereof, having a heavy chain, a light chain, a variable heavy chain domain (VH), a variable light chain domain (VL), and a complementarity determining region (CDR) disclosed herein.

Description

Related applications

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

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

[0003] The field of immuno-oncology has revolutionized the care of cancer patients, improving survival and quality of life. However, to date, much of the research in this field has focused on developing the power of adaptive immune responses through therapeutic targeting of T cells. While these approaches have significantly advanced the field, several challenges remain. Furthermore, the clinical benefits of T-cell therapy cannot be extended to all patients or tumor indications, particularly in the context of solid tumors. Alternative strategies, such as engaging the innate immune system, have become a focus of attention in this field. Specifically, the involvement of natural killer (NK) cells as potent effector cells of the innate immune response has emerged as a promising modality in immunotherapy. Whalen et al. (2023) provided a detailed review of the potential of using NKG2D, CD16A, and other cell surface receptors to engage NK cells in cancer therapy (Whalen et al., mAbs 15:12208697).

[0004] Compared to T cells, natural killer (NK) cells act as the first line of defense, capable of detecting and eliminating tumor- or virus-infected cells without prior antigen-specific activation or differentiation. Therefore, NK cells possess several attractive properties, demonstrating the potential for developing agents that can utilize their therapeutic potential. The intricate balance between activation and inhibition signals, hallmarks of NK cells, requires the involvement of multiple activating receptors, combinations of receptor agonists and NK cell checkpoint inhibitors, and / or combinations with T-cell therapies to maximize therapeutic efficacy in patients. Like T cells, NK cells are increasingly recognized as possessing high cytotoxic potential when appropriately engaged. NK cell therapy, when equipped with potent activating receptors specific to cell surface antigens, has been shown to elicit full patient responses. Similarly, the T-cell receptor complex (TCR) within NK cells enhances 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, it offers the potential for allogeneic, off-the-shelf use, as it avoids graft-versus-host disease (GVHD) caused by a lack of T cell receptors (TCRs). Furthermore, NK cell therapies typically exhibit lower toxicity compared to some T cell-based therapies, including immune effector cell-associated neurotoxicity syndrome (ICAN) and cytokine release syndrome (CRS). However, several key challenges remain with NK cell-based therapies, including inefficient transgene delivery, the complexity of NK cell manufacturing, and the limited depletion and persistence of genetically engineered NK cells.

[0006] Cancer therapy has indeed been revolutionized by developing antibodies that specifically target T cells rather than cancer cells to effectively treat cancer patients and, in some cases, provide durable responses. Unlike cell therapies, mAbs that target T cells, much like NK cell-mediated mAbs, have the potential to reach every effector NK cell, rely on standardized mAb manufacturing methods, and enable predictable pharmacokinetic properties, thus providing potentially potent pan-cancer responses. This field is rich with mAb-based therapies that target the interaction between CD16A / Fc-γ receptor IIIA (FcγRIIIa) expressed on NK cells and the Fcγ 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 (targeting CD38), and trastuzumab (human epidermal growth factor receptor 2-HER2), involve CD16A. However, these previous antibody-based therapies have not utilized another key NK cell activation receptor, NKG2D. NKG2D is a potent regulator of NK cell activation and has recently become an important target in immuno-oncology. NKG2D recognizes eight different ligands upregulated on tumor cells: MHC class I chain-associated protein A (MICA), MHC class I chain-associated protein sequence B (MICB), and UL-16 binding proteins (ULBP), namely ULBP1 through ULBP6.

[0007] Although strategies utilizing NKG2D activity in treatment are still in the early stages of development, MICA exhibits the most consistent and highest expression across all solid tumors and hematologic malignancies compared to other NKG2D ligands. The second most widespread expression pattern is demonstrated by MICB, whose sequence is closely related to MICA. This makes MICA and MICB (MICA / B) attractive pan-cancer targets.

[0008] MICA and MICB are stress-induced surface glycoproteins that are upregulated in various human tumors. MICA / B participate in immune-mediated target cell lysis by activating the receptor NKG2D on NK cells and several T cell subsets.

[0009] However, cancer cells evade NKG2D-mediated NK cell lysis by utilizing the unique α3 region of MICA / B. This region is a target of the extracellular domain (ECD) that is hydrolyzed from cancer cell surface proteins by the action of various proteases expressed on the cell surface or released into the tumor microenvironment by most cancer cells. Therapies are needed to prevent MICA / B shedding, thereby preserving MICA / B on the surface of tumor cells to facilitate NKG2D-mediated NK cell lysis of tumor cells. Such therapies have the potential to treat a variety of different cancers. Therefore, there is an unmet need to develop novel therapies capable of treating cancers that utilize this mechanism to evade immune responses. Summary of the Invention

[0010] This article discloses a method for treating cancer using antibodies that specifically bind to MICA / B.

[0011] Therefore, in a first aspect, this disclosure is characterized by a method of treating a subject with endometrial cancer, comprising administering to the subject a therapeutically effective amount of an antibody that binds to MICA / B (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 not previously 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 pimivalimab, pembrolizumab, nivolumab, cemiplimab, AMP-224, APM-514, or spartalizumab.

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

[0014] In some embodiments of the first aspect, the subject responds 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 has stable disease after administration of the anti-MICA / B antibody. In some embodiments, the response or stable disease 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 implementations 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 includes 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 includes an anti-EGFR antibody. In some embodiments, the anti-EGFR antibody is cetuximab, panitumumab, or necitumumab.

[0017] In some embodiments of the first aspect, the VEGF inhibitor includes 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 includes 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 anti-androgen. 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.

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

[0020] In some implementation schemes of the first aspect, the subjects had metastatic cancer.

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

[0022] In some embodiments of the first aspect, the anti-MICA / B antibody is administered at a dose of about 3 mg / kg to about 10 mg / kg. In some embodiments, the anti-MICA / B antibody is administered to the subject according to a dosing interval. In some embodiments, the dosing interval is referred to as a cycle. In some embodiments, the dosing interval comprises a three-week cycle, wherein the anti-MICA / B antibody is administered once every three 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 repeated dosing interval is performed for 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) and a heavy chain variable region (VH), wherein the VL comprises light chain complementarity-determining region 1 (LCDR1) of SEQ ID NO: 1, light chain complementarity-determining region 2 (LCDR2) of SEQ ID NO: 2, and light chain complementarity-determining region 3 (LCDR3) of SEQ ID NO: 3; and the VH comprises heavy chain complementarity-determining region 1 (HCDR1) of SEQ ID NO: 4, heavy chain complementarity-determining region 2 (HCDR2) of SEQ ID NO: 5, and heavy chain complementarity-determining region 3 (HCDR3) of SEQ ID NO: 6. In some embodiments, the 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 with it. In some embodiments, the 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 with it. 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 with it. 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 with it.

[0024] In some embodiments of the first aspect, the anti-MICA / B antibody specifically binds to the MICA protein, the MICB protein, or both MICA and MICB proteins. In some embodiments, the anti-MICA / B antibody binds to the α-3 domain of the MICA protein, the MICB protein, or both MICA and MICB proteins. 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 intact immunoglobulins, 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 antibody or a chimeric antibody.

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

[0027] In a second aspect, this disclosure is characterized by 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).

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

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

[0030] In some implementations of the second aspect, the cancer is mutated for one or more of the following: EGFR, TP53, HER2, PTEN, ARID1A, CTNNB1, or PLK3R1.

[0031] In some embodiments of the second aspect, the subject responds 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 has stable disease after administration of the anti-MICA / B antibody. In some embodiments, the response or stable disease 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 implementations 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 includes 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 includes an anti-EGFR antibody. In some embodiments, the anti-EGFR antibody is cetuximab, panitumumab, or nexituzumab.

[0034] In some embodiments of the second aspect, the VEGF inhibitor includes 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 includes an anti-VEGF antibody. In some embodiments, the anti-VEGF antibody is bevacizumab or ranibizumab.

[0035] In some embodiments of the second 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 anti-androgen. 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 aspect, the subject has cancer characterized by low levels of PD-L1 expression. In some embodiments, the subject has cancer characterized by high levels of PD-L1 expression. In some embodiments, the subject has cancer characterized by low tumor mutational burden (TMB). In some embodiments, the subject has cancer characterized by high tumor mutational burden (TMB). In some embodiments, the subject has immune-cold cancer. In some embodiments, the subject has immune-hot cancer. In some embodiments, the subject has hormone-sensitive cancer. In some embodiments, the subject has cancer characterized by overexpression of oncogenic drivers. In some embodiments, the subject has cancer expressing one or more of the following: EGFR, ER, PR, or HER2.

[0037] In some implementation schemes of the second aspect, the subjects have metastatic cancer.

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

[0039] In some embodiments of the second aspect, the anti-MICA / B antibody is administered at a dose of about 3 mg / kg to about 10 mg / kg. In some embodiments, the anti-MICA / B antibody is administered to the subject according to a dosing interval. In some embodiments, the dosing interval is referred to as a cycle. In some embodiments, the dosing interval comprises a three-week cycle, wherein the anti-MICA / B antibody is administered once every three 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 repeated dosing interval is performed for 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 comprises a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL comprises light chain complementarity-determining region 1 (LCDR1) of SEQ ID NO: 1, light chain complementarity-determining region 2 (LCDR2) of SEQ ID NO: 2, and light chain complementarity-determining region 3 (LCDR3) of SEQ ID NO: 3; and the VH comprises heavy chain complementarity-determining region 1 (HCDR1) of SEQ ID NO: 4, heavy chain complementarity-determining region 2 (HCDR2) of SEQ ID NO: 5, and heavy chain complementarity-determining region 3 (HCDR3) of SEQ ID NO: 6. In some embodiments, the 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 with it. In some embodiments, the 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 with it. 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 with it. 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 with it.

[0041] In some embodiments of the second aspect, the anti-MICA / B antibody specifically binds to the MICA protein, the MICB protein, or both MICA and MICB proteins. In some embodiments, the anti-MICA / B antibody binds to the α-3 domain of the MICA protein, the MICB protein, or both MICA and MICB proteins. 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 intact immunoglobulins, 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 antibody or a chimeric antibody.

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

[0044] In a third aspect, this disclosure is characterized by a method of treating a subject with cancer, the method 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: (i) a VEGF inhibitor; (ii) an EGFR inhibitor; and / or (iii) hormone therapy.

[0045] In some implementations of the third aspect, the subject responds to a VEGF inhibitor, an EGFR inhibitor, or hormone therapy. In some implementations, the response is a complete response. In some implementations, the response is a partial response. In some implementations, 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.

[0046] In some embodiments of the third aspect, the subject's disease progresses after the response period to a VEGF inhibitor, EGFR inhibitor, or hormone therapy. In some embodiments, the administration of the anti-MICA / B antibody occurs after disease progression following the response period to a VEGF inhibitor, EGFR inhibitor, or hormone therapy.

[0047] In some implementations of the third aspect, the subject has stable disease after administration of a VEGF inhibitor, EGFR inhibitor, or hormone therapy. In some implementations, the subject has a stable disease 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.

[0048] In some implementations of the third aspect, treatment with VEGF inhibitors, EGFR inhibitors, or hormone therapy is discontinued before the administration of anti-MICA / B antibodies is started.

[0049] In some implementation schemes of the third aspect, treatment with VEGF inhibitors, EGFR inhibitors, or hormone therapy is continued after the initiation of anti-MICA / B antibody administration.

[0050] In some implementation schemes of the third aspect, the subject's disease progressed after treatment with VEGF inhibitors, EGFR inhibitors, or hormone therapy.

[0051] In some embodiments of the third aspect, the subject responds 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 has stable disease after administration of the anti-MICA / B antibody. In some embodiments, the response or stable disease 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 includes 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 includes an anti-EGFR antibody. In some embodiments, the anti-EGFR antibody is cetuximab, panitumumab, or nexituzumab.

[0053] In some embodiments of the third aspect, the VEGF inhibitor includes 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 includes an anti-VEGF antibody. In some embodiments, the anti-VEGF antibody is bevacizumab or ranibizumab.

[0054] In some embodiments of the third 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 anti-androgen. 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, the cancer is a female reproductive tract tumor. In some embodiments, the female reproductive tract tumor includes endometrial tumors. In some embodiments, the female reproductive tract tumor includes ovarian cancer or cervical cancer. In some embodiments, the cancer is a salivary gland tumor. In some embodiments, the salivary gland tumor includes mucoepidermoid tumors. In some embodiments, the mucoepidermoid tumor is a parotid gland tumor.

[0056] In some embodiments of the third 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 or 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 adenoid cystic salivary gland tumor. In some embodiments, the subject has peritoneal mesothelioma. In some embodiments, the subject has rectal squamous cell carcinoma. In some embodiments, the subject has leiomyosarcoma. In some embodiments, the subject has colorectal cancer. In some embodiments, the subject has kidney 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 implementations, the subject had head and neck cancer. In some implementations, the subject had appendiceal cancer.

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

[0058] In some implementation schemes of the third aspect, the subjects had metastatic cancer.

[0059] In some implementation schemes of the third aspect, the subject has previously received treatments including chemotherapy.

[0060] In some embodiments of the third aspect, the anti-MICA / B antibody is administered at a dose of about 3 mg / kg to about 10 mg / kg. In some embodiments, the anti-MICA / B antibody is administered to the subject according to a dosing interval. In some embodiments, the dosing interval is referred to as a cycle. In some embodiments, the dosing interval comprises a three-week cycle, wherein the anti-MICA / B antibody is administered once every three 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 repeated dosing interval is performed for 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 comprises a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL comprises light chain complementarity-determining region 1 (LCDR1) of SEQ ID NO: 1, light chain complementarity-determining region 2 (LCDR2) of SEQ ID NO: 2, and light chain complementarity-determining region 3 (LCDR3) of SEQ ID NO: 3; and the VH comprises heavy chain complementarity-determining region 1 (HCDR1) of SEQ ID NO: 4, heavy chain complementarity-determining region 2 (HCDR2) of SEQ ID NO: 5, and heavy chain complementarity-determining region 3 (HCDR3) of SEQ ID NO: 6. In some embodiments, the 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 with it. In some embodiments, the 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 with it. 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 with it. 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 with it.

[0062] In some embodiments of the third aspect, the anti-MICA / B antibody specifically binds to the MICA protein, the MICB protein, or both MICA and MICB proteins. In some embodiments, the anti-MICA / B antibody binds to the α-3 domain of the MICA protein, the MICB protein, or both MICA and MICB proteins. 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 intact immunoglobulins, 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 antibody or a chimeric antibody.

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

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

[0066] In some implementations of the fourth aspect, the response is a complete response. In some implementations, the response is a partial response. In some implementations, 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. In some implementations, the subject's disease progresses after the response period to the PD-1 inhibitor.

[0067] In some implementations of the fourth aspect, the subject has stable disease after administration of a PD-1 inhibitor. In some implementations, the subject has a stable disease 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.

[0068] In some implementations of the fourth aspect, treatment with PD-1 inhibitors is discontinued before the administration of anti-MICA / B antibodies is started.

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

[0070] In some embodiments of aspect four, the subject responds 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 has stable disease after administration of the anti-MICA / B antibody. In some embodiments, the response or stable disease 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 pimiralimumab, pembrolizumab, nivolumab, cimiprimab, AMP-224, APM-514, or spartazolizumab.

[0072] In some implementation schemes 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 includes 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 includes an anti-EGFR antibody. In some embodiments, the anti-EGFR antibody is cetuximab, panitumumab, or nexituzumab.

[0074] In some embodiments of the fourth aspect, the VEGF inhibitor includes 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 includes an anti-VEGF antibody. In some embodiments, the anti-VEGF antibody is bevacizumab or ranibizumab.

[0075] In some embodiments of the fourth 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 anti-androgen. 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, the cancer is a female reproductive tract tumor. In some embodiments, the female reproductive tract tumor includes endometrial tumors. In some embodiments, the female reproductive tract tumor includes ovarian cancer or cervical cancer. In some embodiments, the cancer is a salivary gland tumor. In some embodiments, the salivary gland tumor includes mucoepidermoid tumors. In some embodiments, the mucoepidermoid tumor is a parotid gland tumor.

[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 or 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 adenoid cystic salivary gland tumor. In some embodiments, the subject has peritoneal mesothelioma. In some embodiments, the subject has rectal squamous cell carcinoma. In some embodiments, the subject has leiomyosarcoma. In some embodiments, the subject has colorectal cancer. In some embodiments, the subject has kidney 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 implementations, the subject had head and neck cancer. In some implementations, the subject had appendiceal cancer.

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

[0079] In some implementation schemes of the fourth aspect, the subjects had metastatic cancer.

[0080] In some implementation schemes of the fourth aspect, the subject has previously received treatments including chemotherapy.

[0081] In some embodiments of the fourth aspect, the anti-MICA / B antibody is administered at a dose of about 3 mg / kg to about 10 mg / kg. In some embodiments, the anti-MICA / B antibody is administered to the subject according to a dosing interval. In some embodiments, the dosing interval is referred to as a cycle. In some embodiments, the dosing interval comprises a three-week cycle, wherein the anti-MICA / B antibody is administered once every three 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 repeated dosing interval is performed for 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 comprises a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL comprises light chain complementarity-determining region 1 (LCDR1) of SEQ ID NO: 1, light chain complementarity-determining region 2 (LCDR2) of SEQ ID NO: 2, and light chain complementarity-determining region 3 (LCDR3) of SEQ ID NO: 3; and the VH comprises heavy chain complementarity-determining region 1 (HCDR1) of SEQ ID NO: 4, heavy chain complementarity-determining region 2 (HCDR2) of SEQ ID NO: 5, and heavy chain complementarity-determining region 3 (HCDR3) of SEQ ID NO: 6. In some embodiments, the 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 with it. In some embodiments, the 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 with it. 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 with it. 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 with it.

[0083] In some embodiments of the fourth aspect, the anti-MICA / B antibody specifically binds to the MICA protein, the MICB protein, or both MICA and MICB proteins. In some embodiments, the anti-MICA / B antibody binds to the α-3 domain of the MICA protein, the MICB protein, or both MICA and MICB proteins. 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 intact immunoglobulins, 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 antibody or a chimeric antibody.

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

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

[0087] In some embodiments of aspect five, the corticosteroid is administered 30-60 minutes prior to the administration of the anti-MICA / B antibody. In some embodiments, the corticosteroid is administered at a dose of about 2 mg to about 50 mg. In some embodiments, the corticosteroid is administered at 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 the anti-MICA / B antibody.

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

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

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

[0092] In some embodiments of aspect five, 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 or 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 adenoid cystic salivary gland tumor. In some embodiments, the subject has peritoneal mesothelioma. In some embodiments, the subject has rectal squamous cell carcinoma. In some embodiments, the subject has leiomyosarcoma. In some embodiments, the subject has colorectal cancer. In some embodiments, the subject has kidney 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 implementations, the subject had head and neck cancer. In some implementations, the subject had appendiceal cancer.

[0093] In some embodiments of aspect five, the subject has cancer characterized by low levels of PD-L1 expression. In some embodiments, the subject has cancer characterized by high levels of PD-L1 expression. In some embodiments, the subject has cancer characterized by low tumor mutational burden (TMB). In some embodiments, the subject has cancer characterized by high tumor mutational burden (TMB). In some embodiments, the subject has immune-cold cancer. In some embodiments, the subject has immune-hot cancer. In some embodiments, the subject has hormone-sensitive cancer. In some embodiments, the subject has cancer characterized by overexpression of oncogenic drivers. In some embodiments, the subject has cancer expressing one or more of the following: EGFR, ER, PR, or HER2.

[0094] In some implementation schemes of the fifth aspect, the subjects had metastatic cancer.

[0095] In some implementation schemes of the fifth aspect, the subject has previously received treatment including chemotherapy.

[0096] In some embodiments of aspect five, the anti-MICA / B antibody is administered at a dose of about 3 mg / kg to about 10 mg / kg. In some embodiments, the anti-MICA / B antibody is administered to the subject according to a dosing interval. In some embodiments, the dosing interval is referred to as a cycle. In some embodiments, the dosing interval comprises a three-week cycle, wherein the anti-MICA / B antibody is administered once every three 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 repeated dosing interval is performed for 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 comprises a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL comprises light chain complementarity-determining region 1 (LCDR1) of SEQ ID NO: 1, light chain complementarity-determining region 2 (LCDR2) of SEQ ID NO: 2, and light chain complementarity-determining region 3 (LCDR3) of SEQ ID NO: 3; and the VH comprises heavy chain complementarity-determining region 1 (HCDR1) of SEQ ID NO: 4, heavy chain complementarity-determining region 2 (HCDR2) of SEQ ID NO: 5, and heavy chain complementarity-determining region 3 (HCDR3) of SEQ ID NO: 6. In some embodiments, the 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 with it. In some embodiments, the 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 with it. 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 with it. 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 with it.

[0098] In some embodiments of the fifth aspect, the anti-MICA / B antibody specifically binds to the MICA protein, the MICB protein, or both MICA and MICB proteins. In some embodiments, the anti-MICA / B antibody binds to the α-3 domain of the MICA protein, the MICB protein, or both MICA and MICB proteins. 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 intact immunoglobulins, 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 antibody or a chimeric antibody.

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

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

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

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

[0104] In some embodiments, the method further includes administering a therapeutically effective amount of corticosteroid to the subject prior to the first dose of the anti-MICA / B antibody. In some embodiments, the corticosteroid is administered 30-60 minutes prior to the administration of the anti-MICA / B antibody. In some embodiments, the corticosteroid is administered at a dose of about 2 mg to about 50 mg. In some embodiments, the corticosteroid is administered at a dose of about 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, the corticosteroid is administered before the first 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. Attached Figure Description

[0106] The patent or application document contains at least one color drawing. A copy of the publication of this patent or patent application with one or more color drawings will be provided by the Patent Office upon request and after payment of the necessary fees.

[0107] An understanding of the features and advantages of the invention can be obtained by referring to the following illustrative embodiments, which utilize the principles of the invention, and wherein the accompanying drawings are:

[0108] Figure 1A-1C The binding characteristics of CLN-619 are described. Figure 1A The average of three independent experiments showing the binding of CLN-619 to a representative MICA allele and a typical MICB allele, as measured by Octet, is displayed. Figure 1B The mean of two independent experiments showing CLN-619 binding to representative MICA alleles and typical MICB alleles, as measured by ELISA, is shown. (N=2). CLN-619 has similar binding levels with a wide range of proteins, so not all symbols are visible. (c) In two independent experiments, CLN-619 bound to 28 of the most common MICA alleles, as measured by Luminex. 6D4 was used as a positive control (PMID: 28334733). Error bars represent SEM.

[0109] Figure 2The CLN-619 epitope was depicted. The X-ray crystal structure of the human Fab antibody fragment CLN-619 complexed with the antigen MICA1 α3 domain was obtained at a resolution of 2.12 Å. The binding interface of CLN-619 is a discontinuous epitope containing only 19 amino acid residues from the MICA1 α3 domain.

[0110] Figure 3A and 3B This demonstrates that CLN-619 regulates cell surface MICA / B by inhibiting shedding. Figure 3A This is a graph showing the cell surface levels of MICA / B as measured by flow cytometry. Figure 3B This is a graph showing the soluble MICA / B ratio in cell supernatant as measured by ELISA (N=3 or N=2; error bars represent SEM). Geometric mean EC50 values ​​and 95% confidence intervals are reported.

[0111] Figure 4A and 4B This demonstrates that CLN-619 mediates NK cell activation and cell killing of target cells expressing MICA / B. Figure 4A This is a graph from an ELISA assay used to measure IFNγ in cell supernatant collected from a 96-hour co-culture killing assay of NK cells and target cells expressing MICA / B (E:T 40:1) in the presence of serially titrated antibodies. N=3. CLN-619 DANA is a mutant (D265A / N297A), Fc-silenced form of CLN-619. Figure 4B This graph is from the xCELLigence platform, which is used for measurement. Figure 4A The experiment described above showed the death of target cells expressing MICA / B. Representative curves are shown.

[0112] Figure 5 This study demonstrated CLN-619-induced ADCC. MICA-expressing target cells treated with gradient-titrated antibody were co-cultured with NFAT-luciferase Jurkat cells expressing either a high-affinity (V158) or low-affinity (F158) variant of human FcγRIIIa (CD16A) at an E:T ratio of 10:1. Luciferase activity was quantified after 6 hours. Geometric mean EC50 values ​​and 95% confidence intervals are reported, after subtracting E:T control background.

[0113] Figure 6This study demonstrates that CLN-619 promotes antibody-dependent phagocytosis. Phagocytosis of fluorescently labeled target cells by macrophages was measured by flow cytometry. Representative curves of HCC1534 and HCT-116 cells treated with CLN-619 from two donors are presented. Cetuximab and allotype control antibody treatments were performed at the highest concentrations. Geometric mean EC50 values ​​and 95% confidence intervals are reported. E:T control background is subtracted. Error bars represent SEM.

[0114] Figure 7A –7C indicates that CLN-619 treatment elicited a potent antitumor effect in a preclinical mouse model. Figure 7A This is a graph depicting the results in BALB / c SCID mice inoculated with PLC / PRF / 5 human hepatocellular carcinoma cells (N=10 / group). Mice were treated with two dose levels of CLN-619 three times weekly via intraperitoneal injection. Statistical data represent one-way ANOVA multiple comparisons. *P=0.015. Figure 7B This is a graph depicting the results in BALB / c SCID mice inoculated with HCC1534 human lung cancer cells (N=10 / group). Mice were treated with a series of doses of CLN-619 or Fc-silenced CLN-619 twice weekly via intraperitoneal injection. Statistical data represent univariate ANOVA multiple comparisons. NS - not significant. Figure 7C This is a graph depicting the results in BALB / c SCID mice inoculated with HCT-116 luciferase-labeled human colon cancer cells (N=10 / group). Mice were treated with CLN-619 or hIgG1 twice weekly via intraperitoneal injection. Survival was analyzed using the Kaplan-Meier assay on day 41.

[0115] Figure 8 This is a table showing the allele frequencies of MICA allele variants tested in luminex assays. Allele frequencies are based on high-throughput sequencing of more than 2 million donor samples, as previously described

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

[0116] Figure 9 This table displays the binding of CLN-619 to FcRs. Kinetic binding parameters were determined using Biacore-SPR. The Biacore chip was coupled to various His-labeled FcRs using anti-HIS antibodies, and the bound FcRs were then detected by test antibodies. Kinetic analysis was performed using Biacore software. Relative scale of affinity (M): ++++ 10 -8 to -9 ;+++ 10 -7 ;++ 10 -6 +10-5 + / - Detectable bindings; - Undetectable bindings.

[0117] Figure 10 This table shows the sequence analysis of the CLN-619 epitope to the MICA / B alleles. The listed alleles have a frequency >1% in the population and are highly homologous to the conserved amino acid changes shown in orange. * indicates a typical allele.

[0118] Figure 11 This diagram illustrates that CLN-619 does not interfere with MICA-NKG2D binding. Primary NK cells were incubated with His-labeled MICAECD in the presence of CLN-619 or hIgG1 control antibodies, either without (a) or containing anti-NKG2D antibodies. The amount of MICA bound to the cell surface was measured by flow cytometry.

[0119] Figure 12 This graph illustrates the ability of CLN-619 DANA to retain the regulatory capacity for MICA / B on the cell surface. Cell surface levels of MICA / B were measured by flow cytometry.

[0120] Figure 13A and 13B The pharmacokinetic parameters of CLN-619 following a single IP injection in BALB / c SCID mice were described. Figure 13A This describes the concentration-time curves and pharmacokinetic parameters (e.g., after a single IP administration of 1, 3, and 10 mg / kg of CLN-619) Figure 13B The graph shown is presented. Report the mean and standard deviation.

[0121] Figure 14 This is a table describing the baseline characteristics of patients recruited to the Phase 1 open-label dose-escalation study of CLN-619 for the treatment of advanced solid tumors.

[0122] Figure 15 This is a schematic diagram illustrating the structure of a dose escalation study, including the number of patients in each dose level group.

[0123] Figure 16 This is a table summarizing treatment-related adverse events (TEAEs) observed in ≥ 10% of patients.

[0124] Figure 17A and 17B These are graphs and tables showing the mean serum concentration of CLN-619 at different time points after administration and the PK parameters stratified by dose level.

[0125] Figure 18This is a series of graphs showing cytokine expression in longitudinal serum samples from patients treated with CLN-619 monotherapy using the Myriad RBM Inflammation and custom MAP assays on the Luminex platform. The graphs depict the mean absolute cytokine levels (pg / ml) at each dose level over time. Error bars represent SD.

[0126] Figure 19A and 19B This is a graph showing the treatment time and clinical activity of each study patient who received CLN-619 monotherapy.

[0127] Figure 20A and 20B This is a table summarizing the objective responses and best monotherapy responses of patients in studies who received CLN-619 monotherapy.

[0128] Figure 21 This is a series of images of a study patient with mucoepidermoid parotid gland carcinoma at baseline (top), during the 4th cycle of CLN-619 treatment (middle), and during the 7th cycle of CLN-619 treatment (bottom).

[0129] Figure 22 Images of a study patient with endometrial cancer at baseline (left) and at the 10th cycle of CLN-619 treatment (right).

[0130] Figure 23A and 23B The overall study design of clinical studies of CLN-619 alone and in combination with pembrolizumab in patients with advanced solid tumors is shown.

[0131] Figure 24 The event schedule for clinical studies of CLN-619 alone and in combination with pembrolizumab in patients with advanced solid tumors is shown.

[0132] Figure 25 The table shows the sampling schedule for clinical studies of CLN-619 alone and in combination with pembrolizumab in patients with advanced solid tumors.

[0133] Figure 26A The infusion time description for CLN-619-001 is displayed.

[0134] Figure 26B The infusion time description for module D (load dose group) CLN-619-001 is displayed.

[0135] Figure 27 The treatment time and clinical activity of the combined groups are displayed.

[0136] Figure 28The treatment duration and clinical activity of patients with NSCLC are shown.

[0137] Figure 29 Treatment duration and clinical activity are shown for monotherapy groups of ≥1 mg / kg.

[0138] Figure 30 This is an overview of treatment-related adverse events (TEAEs).

[0139] Figure 31 The overall study design for using CLN-619 to treat relapsed / relapsed multiple myeloma is shown.

[0140] Figure 32A and 32B The effect of CLN-619 on T cell activation was demonstrated. Figure 32A The image is from the xCELLigence platform, which measures cell lysis in a 72-hour co-culture kill assay using negatively selected CD8+ T cells from PBMCs combined with target cells expressing MICA / B (E:T 10:1) in the presence of continuously titrated antibodies. Figure 32B This is a graph from flow cytometry, which is used to measure data from... Figure 32A The percentage of CD8+ T cells expressing CD25 in the experiment. Figure 32C The effect of CLN-619 on T cell lysis was shown. Measurements were taken using ELISA. Figure 32A The experiment described in the paper involves the collection of IFNγ in the cell supernatant. Figures 32A-32C N=3. A representative curve is shown. Detailed Implementation

[0141] In some embodiments, this document discloses monoclonal antibodies that specifically bind to MICA / B. In some embodiments, the MICA / B antibodies of this document bind to the MICA / B protein or fragments thereof and modulate an individual's immune response, thereby treating cancer (e.g., hepatocellular carcinoma).

[0142] Major histocompatibility complex class I related chains A and B (MICA / B) are two stress-inducible ligands of the natural killer (NK) cell receptor NKG2D, playing a crucial role in mediating the cytotoxicity of NK and T cells. Soluble MICA / B shed from diseased cells (e.g., cancer cells) desensitizes NK and T cells via binding to the NKG2D receptor, thereby suppressing the immune response. Therefore, modulating MICA / B helps regulate the immune response in individuals (e.g., those with cancer).

[0143] The development of therapeutics that inhibit MICA / B shedding is an emerging strategy utilizing these NKG2D ligands. The clinical data presented in this article demonstrate the safety and efficacy of targeting and inhibiting MICA / B shedding using CLN-619 (a humanized IgG1 antibody, currently the only clinically available mAb targeting MICA / B). CLN-619 prevents MICA / B shedding from cancer cells by binding to the α3 domain and contains the active Fcγ1 domain to drive ADCC and ADCP. The MICA / B gene is highly polymorphic; however, CLN-619 exhibited surprisingly broad reactivity to all tested allele variants. Reduced levels of shed MICA and an associated increase in MICA on the surface of tumor cells have been observed with CLN-619 treatment. Importantly, the α1 and α2 domains of MICA binding to NKG2D are not blocked by the binding antibody. In fact, CLN-619 enhanced the binding of MICA to NKG2D, which appears to be dependent on the antibody's Fcγ1 domain, which can simultaneously bind to CD16A on NK cells. Notably, CLN-619 exhibited significant single-agent activity at low doses in tumor xenograft models, with its activity primarily dependent on the functional Fcγ1 domain. Therefore, CLN-619 holds promise for therapeutic efficacy as a single agent or in combination with other agents that engage other NK cell receptors and / or one or more checkpoint inhibitors.

[0144] The stabilizing effect of CLN-619 on MICA / B leads to the accumulation of these NKG2D ligands on the surface of tumor cells, thereby overcoming immune evasion caused by MICA / B shedding. Unbound by any specific theory or mechanism of action, this may be due to the increased target antigen density, maximizing the antibody's ADCC and ADCP functions. Importantly, CLN-619 has the potential to activate NK cells by simultaneously and potentially synergistically engaging two key activating receptors, NKG2D and CD16A. Published data support the cooperation between these two receptors, indicating that NKG2D engagement can lower the activation threshold of several receptors, including CD16A. Given that NK cell activation receptors are frequently downregulated in the TME, the cooperation between these two receptors may be important. This publication presents important data from a Phase 1 clinical trial (NCT05117476) of CLN-619 in patients with advanced solid tumors treated alone or in combination with pembrolizumab.

[0145] As demonstrated by a retrospective analysis, monotherapy activity is uncommon in phase 1 oncology studies, which showed single-digit objective response rates in over 4,000 patients enrolled in phase 1 solid tumor studies sponsored by the National Cancer Institute between 2000 and 2019. CLN-619's monotherapy response rate in evaluable patients is superior to historical NCI data as well as TIGIT and lag-3 antibodies, and is indeed encouraging in the early stages of the CLN-619 program.

[0146] Further details regarding clinical trials of CLN-619 in patients with advanced solid tumors, either alone or in combination with pembrolizumab, can be found in the appendix.

[0147] One such study was a phase 1 dose-escalation and expansion study that tested CLN-619 monotherapy and CLN-619 in combination with pembrolizumab in staggered parallel cohorts. CLN-619 monotherapy was administered every 3 weeks. A prophylactic drug was required only before the first infusion and was mandated to include a corticosteroid starting at a dose level of 3 mg / mg. Dose-limiting toxicities were evaluated during the first cycle, and response was assessed every 3 cycles according to RECIST criteria.

[0148] The monotherapy group received a standard 3x3 dose escalation, testing doses from 0.1 to 10 mg / mg. Once the dose level was cleared, the dose level targeting DLT clearance could be further extended (sometimes referred to as “backfilling”), treating up to 10 patients per dose. A total of 37 patients were treated, with a median age of 63 years and 62% being female. This large pre-treatment group had received a median of 3 prior therapies, and just over half had received CPI therapy.

[0149] RECIST responses were observed starting at doses of 3 mg / kg, including sustained CR in parotid patients treated with 3 mg / kg and two PRs in endometrial patients treated with 3 and 10 mg / kg. One of the parotid CRs and one of the endometrial PRs occurred in patients whose disease progressed during previous checkpoint inhibitor therapy. At doses of 1 mg / kg and above, 10 of the 22 evaluable response patients achieved an objective response or optimal response with stable disease.

[0150] In addition to two cases of endometrial partial response (PR), three cervical patients achieved stable disease, and two ovarian patients also achieved stable disease. Data shows that among the 10 patients with gynecological cancer recruited to the study, 7 achieved an objective response or a stable disease response (best response).

[0151] Durable stable disease was also observed in one breast cancer patient and one salivary gland cancer patient, highlighting objective clinical activity of monotherapy across multiple tumor types.

[0152] In a scan from a patient with a parotid gland tumor who achieved a sustained complete remission (CR), biopsy-confirmed lymph node recurrence met the criteria for a complete response at C4D1, was confirmed at C7D1, and persisted beyond the data cutoff of more than 9 months after initiation of CLN-619 treatment. Notably, this patient had experienced 30 months of sustained partial response (PR) on checkpoint inhibitor therapy prior to disease progression during treatment.

[0153] In a scan of one of two endometrial patients who experienced confirmed partial remission (PR), baseline images showed multiple bilateral lung metastases that had regressed to the PR criteria at C4D1 and were confirmed by subsequent scans at C7D1. This patient had experienced disease progression after prior therapy with platinum-based chemotherapy plus Herceptin, anastrozole, and doxorubicin, and had recently received 9 months of treatment with pembrolizumab plus lenvantinib.

[0154] Adverse events occurring in >10% of patients demonstrate that most AEs observed during CLN-619 monotherapy were Grade 1 / 2. Notably, no dose-limiting toxicities were observed at any assessed dose level. A single Grade 3 treatment-related AE, laryngeal edema, occurred during the first infusion in a patient who had not received a mandatory steroid prodrug; this is an infusion-related adverse event observed when using other monoclonal antibodies. All other IRRs were Grade 1 or 2 and occurred only in the first cycle. The data support the view that CLN-619 monotherapy is well-tolerated, and therefore its safety profile should support easy conjugation with other agents.

[0155] The data further demonstrate that CLN-619 monotherapy is well tolerated, with no DLT at up to 10 mg / kg, and that CLN-619 has shown significant monotherapy activity in a variety of tumor types, including patients whose disease progressed during checkpoint inhibitor therapy, including confirmed and sustained objective responses.

[0156] Given the widespread expression of this pathway in tumors, it is hypothesized that CLN-619 may have pan-cancer therapeutic activity.

[0157] The data presented in this embodiment 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-unexperienced patients. Of particular note is the significant single-agent activity in multiple gynecological malignancies, such as endometrial and cervical cancer. Clinical activity in multiple gynecological cancer types demonstrates the potential of CLN-619 in patients with high unmet needs. More specifically, dose escalation of monotherapy demonstrated the acceptable safety profile of CLN-619 at 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, infusion-related responses were grade 1 / 2 and limited to the first dose in patients receiving a mandatory prophylactic agent. Importantly, CLN-619 therapy is effective against multiple tumor types at the administered doses, including demonstration of stable disease observed in tumors in diverse patients, including cervical cancer, ovarian cancer, salivary gland cancer, and breast cancer. Some patients had a median of 3 prior systemic therapies (1–7), including 54% who had previously received immune checkpoint inhibitors. First-ever clinical data on CLN-619 monotherapy demonstrate its broad potential across multiple tumor types. Therefore, CLN-619 has the potential to treat patients with extensive tumor recurrence or who are ineligible for checkpoint inhibitor therapy.

[0158] Further data showed that the combination therapy of CLN-619 and pembrolizumab was well tolerated and demonstrated meaningful clinical activity, including objective response, in tumor types that typically do not respond to pembrolizumab.

[0159] Data from CLN-619-001 (NCT05117476) are described in the examples. This is the first human dose escalation and dose expansion study of CLN-619 alone or in combination with anti-PD1 in patients with advanced solid tumors. 64 patients received either CLN-619 in combination with pembrolizumab (n=22) or as monotherapy (n=42).

[0160] Monotherapy efficacy has been observed in a variety of cancers, including confirmed responses at doses of 3 mg / kg and above. CLN-619 remains tolerable at doses up to 10 mg / kg, and no dose-limiting toxicities were observed at any of the tested doses. In some embodiments, CLN-619 monotherapy is administered intravenously every three weeks.

[0161] Pharmacokinetic (PK), pharmacodynamic (PD), and mechanism of action studies were conducted to observe the PK and PD effects of CLN-619 in the surrounding and tumor microenvironment.

[0162] PK analysis of CLN-619 monotherapy dose escalation 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, as measured by Cmax, was observed between 0.1 and 10 mg / kg. Furthermore, a greater-than-dose-proportional increase in CLN-619 exposure, as measured by AUC0–504h, was observed between 0.1 and 1 mg / kg, and a nearly dose-proportional increase in CLN-619 exposure was observed between the 1 and 10 mg / kg dose levels.

[0163] The study investigated cytokine expression in longitudinal serum samples from patients treated with CLN-619 monotherapy. Data showed a transient increase in cytokine levels observed 2–6 hours after the first dose of CLN-619, which returned to baseline within 4–8 days. No dose-dependent increase in peripheral cytokine secretion was observed.

[0164] Increased MICA / B expression within tumors was observed after treatment. Analysis of nine paired biopsies using MultiplexIF and quantification using an image analysis algorithm (Flagship Biosciences) revealed increased MICA / B positivity in most paired biopsies from patients who received treatment (cycle 2, day 8) compared to baseline.

[0165] The localization of the MICA / B membrane in nine paired biopsies was also analyzed by IHC and quantified by pathology as the percentage of membrane-stained positive cells. The results showed that the MICA / B membrane positivity rate was increased in most treated biopsies.

[0166] The data in the appendix show that monotherapy activity was achieved in patients who had received substantial prior treatment, including after progression on anti-PD1 therapy. Objective responses to CLN-619 monotherapy were observed in multiple tumor types, including after progression on checkpoint inhibitor therapy and significant monotherapy activity observed in gynecological malignancies.

[0167] The data in the examples also show two case studies of subjects with endometrial tumors. In the first case study, the subject had endometrioid endometrial cancer and showed a confirmed partial response after 3 cycles of CLN-619. This subject had received 3 prior therapies and had not received checkpoint inhibitor intervention. In the second case study, the subject had serous endometrial cancer and showed a confirmed partial response after 6 cycles of CLN-619. This subject had received five prior therapies, including pembrolizumab and levantinib. The data showed that reactive endometrial tumors have microsatellite stability, low tumor mutational burden, and low neoantigen presentation index.

[0168] The tumor microenvironment of patients with stable disease for more than 6 months was analyzed using paired biopsies from 13 patients treated at dose levels of 3, 6, and 10 mg / kg in the dose-level extension cohort. Data showed that tumors with long-term stable disease had lower tumor mutational burden and lower neoantigen presentation. In tumors from patients with stable disease for more than 6 months, MICA / B expression and NK cell activation increased after CLN-619 treatment. In 2 / 3 of the paired biopsies, PDL1 expression and C8 activation increased after treatment.

[0169] The study also showed that soluble MICA (sMICA) in patient serum is a PD biomarker for CLN-619. In this study, endogenous IgG in negative serum samples containing recombinant sMICA or normal serum was pre-cleaned using Protein G Sepharose beads. The supernatant after bead removal was immunoprecipitated with hIgG1 or CLN-619. sMICA remaining in the supernatant after immunoprecipitation was quantified by ELISA and plotted as a percentage of the control. The study showed that CLN-619 binds to sMICA in human serum. Further, CLN-619 treatment increased serum sMICA levels, and sMICA levels are a surrogate indicator of CLN-619-induced target involvement. This was demonstrated by modeling CLN-619 drug concentrations to predict sMICA levels. Furthermore, no sedimentation effect was observed. The CLN-619 content in patient blood was found to be approximately 1000 times higher than the sMICA level, indicating that only a small fraction of CLN-619 binds to sMICA at any given time. No clinical relevance was observed in the binding of this small subset of CLN-619 to sMICA.

[0170] Therefore, consistent with the proposed mechanism of action of CLN-619, membrane localization of MICA / B increases after treatment with CLN-619. Thus, this invention demonstrates a method for increasing MICA / B localization in a subject by administering an effective amount of CLN-619.

[0171] In some embodiments, the presence of sMICA is demonstrated as a pharmacodynamic biomarker for CLN-619. The method of the present invention includes monitoring sMICA levels in the presence and absence of CLN-619 therapy, wherein an increase in sMICA presence indicates that CLN-619 is therapeutically effective. Data presented herein show that the increase in sMICA levels is dose-responsive and depends on the dose of CLN-619 administered.

[0172] Other methods described herein use sMICA as a measure of targeting engagement induced by CLN-619. These methods, for example, relate to methods for determining the effectiveness or efficacy of CLN-619 in subjects, including determining the presence of sMICA in the serum of the subjects before and after administration of CLN-619, wherein an increase in the presence of sMICA indicates said effectiveness or efficacy.

[0173] Furthermore, data demonstrating objective responses and long-term stable disease (6 months or longer) in tumors with low TMB and low antigen presentation index support the potential of CLN-619 to elicit a response independent of a skilled antigen presentation mechanism. This is consistent with the proposed multimodal mechanism of action of CLN-619. Therefore, this paper presents a method for treating solid tumors with CLN-619, wherein the treatment results in at least a partial response of the tumor to the CLN-619. The method may also include demonstrating disease remission or stabilization over a period of at least six months. Therefore, the method of the present invention can also be used to extend the lifespan of a subject by at least six months using the compositions of the present invention. These and other aspects of the invention will be further detailed below.

[0174] definition

[0175] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains.

[0176] As used herein, “MICA / B” refers to MICA protein, MICB protein, or MICA and MICB proteins, including variants, isotypes, and species homologs of human MICA / B.

[0177] As used herein, an "antibody" refers to a glycoprotein that exhibits binding specificity to a particular antigen. Antibodies typically contain variable and constant domains in each of the heavy and light chains. Thus, most antibodies have a heavy chain variable domain (VH) and a light chain variable domain (VL), which together form part of the antibody that binds to the antigen. Within each variable domain are three complementarity-determining regions (CDRs) that form loops within the heavy chain variable domain (VH) and light chain variable domain (VL), contacting the surface of the antigen. Antibodies in this article also include the complete molecule and its functional fragments, also referred to as the "antigen-binding moiety" or the fragment of the antibody capable of binding to an antigen.

[0178] As used herein, a “chimeric” antibody is an antibody in which a portion of its heavy and / or light chain is identical or homologous to a corresponding sequence in an antibody derived from a particular species or belonging to a particular class or subclass of antibodies, while the remainder of the chain is identical or homologous to a corresponding sequence in an antibody derived from another species or belonging to another class or subclass of antibodies, and fragments of such antibodies, provided they exhibit the desired biological activity (see, for example, Morrison et al., Proc. Natl. Acad. Sci. USA 81:6851-6855 (1984)). In this context, “humanized antibody” refers to a chimeric antibody with human sequence substitutions in its antibody sequence.

[0179] The terms “recipient,” “individual,” “subject,” “host,” and “patient” are used interchangeably herein and, in some cases, refer to any mammalian subject for whom a diagnosis, treatment, or therapy is desired, specifically a human. “Mammalian” for therapeutic purposes means any animal classified as a mammal, including humans, livestock, and farm animals, as well as laboratory animals, zoo animals, sporting animals, or pet animals, such as dogs, horses, cats, cattle, sheep, goats, pigs, mice, rats, rabbits, guinea pigs, monkeys, etc. In some implementations, the mammal is a human.

[0180] As used herein, the term "treatment / treating" in some contexts refers to the administration of an agent or the performance of a procedure to achieve a certain effect. An effect may be preventative in relation to the complete or partial prevention of a disease or its symptoms and / or therapeutic in relation to the partial or complete cure of a disease and / or its symptoms. As used herein, "treatment" may include treating a disease or condition (e.g., cancer) in mammals, particularly in humans, and includes: (a) preventing the occurrence of a disease or its symptoms in subjects who may be susceptible to the disease but have not yet been diagnosed with it (e.g., including diseases that may be associated with or caused by a primary disease); (b) suppressing the disease, i.e., halting its progression; and (c) alleviating the disease, i.e. causing its remission. Treatment refers to any successful marker of treating or improving or preventing cancer, including any objective or subjective parameter such as reduction; relief; weakening of symptoms or making the disease condition more tolerable to the patient; slowing the rate of degeneration or decline; or reducing the debilitating nature of a degenerative endpoint. Treatment or improvement of symptoms is based on one or more of objective or subjective parameters; including the results of a physician's examination. Therefore, the term "treatment" includes the administration of the compounds or agents of the present invention to prevent or delay, alleviate, or inhibit the development of symptoms or ailments associated with a disease (e.g., cancer). The term "therapeutic effect" refers to the reduction, elimination, or prevention of a subject's disease, symptoms of a disease, or side effects of a disease.

[0181] "Therapeutic effective amount" in some contexts means the amount that, when administered to a subject, is sufficient to treat the disease.

[0182] As used herein, unless the context clearly indicates otherwise, the singular forms “a,” “and,” and “the” include plural indicators. Thus, for example, references to “an antibody” include multiple antibodies.

[0183] As used in this article, “about” a number means a range including that number and from 10% below that number to 10% above that number. “About” a range means from 10% below the lower limit of the range to 10% above the upper limit of the range.

[0184] "Percentage (%) identity" refers to the degree to which two sequences (nucleotides or amino acids) have the same residues at the same position when aligned. For example, "an amino acid sequence has X% identity with SEQ ID NO:Y" means that the amino acid sequence is identical to SEQ ID NO:Y by a percentage, specifically stating that X% of the residues in the amino acid sequence are identical to the residues in the sequence disclosed in SEQ ID NO:Y. Such calculations are generally performed using computer programs. Exemplary programs for comparing and aligning sequence pairs include ALIGN (Myers and Miller, 1988), FASTA (Pearson and Lipman, 1988; Pearson, 1990), and nick BLAST (Altschul et al., 1997), BLASTP, BLASTN, or GCG (Devereux et al., 1984).

[0185] MICA / B

[0186] In some embodiments, this document discloses monoclonal antibodies that specifically bind to MICA / B. In some embodiments, this document further discloses monoclonal antibodies that competitively bind to MICA / B.

[0187] Major histocompatibility complex (MHC) class I chain-associated gene A and gene B proteins (MICA / B) are glycosylated, polymorphic, and membrane-anchored non-classical MHC class I proteins. MICA / B is associated with MHC class I and has a similar domain structure, containing three extracellular Ig-like domains (α-1, α-2, and α-3), a transmembrane domain, and a C-terminal cytoplasmic tail. MICA / B is a ligand for the C-type lectin-like activated receptor natural killer cell family 2D (NKG2D) on immune effector cells, including NK, NKT, and αβ and γδ CD8+ T cells. The interaction between MICA / B and NKG2D plays a role in tumor surveillance and immune responses.

[0188] MICA / B proteins are normally expressed at low levels in normal cells, but are induced to higher levels in stressed or transformed cells (e.g., cancer cells). The interaction between immune effector cells carrying NKG2D and stressed or diseased cells expressing MICA / B ligands on their cell surfaces generates a cellular immune response against the stressed / diseased cells, ultimately leading to the death of MICA / B-expressing cells. In cancer cells, truncated MICA / B proteins (proteins lacking a transmembrane domain and cytoplasmic tail but retaining three extracellular domains including α-1, -2, and -3 domains) are frequently shed into the bloodstream by proteases, leading to the downregulation (receptor internalization) of their intended receptor, NKG2D, on effector immune cells. In some cases, MICA / B glycoproteins are produced intracellularly; they typically do not bind to the cell surface membrane but are instead incorporated into exosomes and released extracellularly, where they interact with NKG2D receptors on immune cells. These truncated or soluble MICA / B ligands, detached from the surface of cancer cells, act as decoy molecules, leading to the downregulation of NKG2D receptors on immune effector cells such as NK, NKT, and various CD8+ T cells. In some cases, the formation of soluble MICA / B can result in an unusual situation where effector molecules of the innate defense system (whose natural function is to seek out and destroy transformed cells) are shut down by the immunosuppressive effects of these decoy ligand molecules, allowing cancer cells to evade the immune system and grow uncontrollably.

[0189] Anti-MICA / B antibody

[0190] This document provides antibodies that specifically bind to the MICA / B protein. 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, VL, and CDRs determine the antigen-binding specificity and antigen-binding strength of the antibody. The amino acid sequences of the heavy and light chains, VH and VL, and CDRs are summarized in Table 1.

[0191] Table 1: Anti-MICA / B Monoclonal Antibody Sequences

[0192]

[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 binds to the α-3 domain of the MICA protein. In some embodiments, the antibody binds to the α-3 domain of the MICB protein. In some embodiments, the antibody binds to the α-3 domain of both the MICA and MICB proteins. In some embodiments, the antibody binds to the MICA protein as a membrane-bound MICA protein. In some embodiments, the antibody binds to the MICA protein as a soluble MICA protein. In some embodiments, the antibody binds to the MICA protein as both a membrane-bound and soluble MICA protein. In some embodiments, the antibody binds to the MICB protein as a membrane-bound MICB protein. In some embodiments, the antibody binds to the MICB protein as 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 intact immunoglobulins, scFv, Fab, F(ab')2, or disulfide-linked 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 complementarity-determining region

[0196] This document discloses antibodies that specifically bind to MICA / B, said antibodies having a light chain comprising a light chain complementarity-determining region (CDR). In some embodiments, the antibody binding to MICA / B comprises a light chain CDR sequence having an amino acid sequence having at least about 70% identity with at least one of the amino acid sequences shown in SEQ ID NO: 1-3. In some embodiments, the antibody binding to MICA / B comprises a light chain CDR sequence having an amino acid 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% identity with at least one of the amino acid sequences shown in SEQ ID NO: 1-3. In some embodiments, the antibody binding to MICA / B comprises a light chain CDR sequence having an amino acid sequence having 100% identity with at least one of the amino acid sequences shown in SEQ ID NO: 1-3.

[0197] This document further discloses antibodies that specifically bind to MICA / B, said antibodies having a heavy chain comprising a heavy chain complementarity-determining region (CDR). In some embodiments, the antibody binding to MICA / B comprises a heavy chain CDR sequence having an amino acid sequence having at least about 70% identity with at least one of the amino acid sequences shown in SEQ ID NO: 4-6. In some embodiments, the antibody binding to MICA / B comprises a heavy chain CDR sequence having an amino acid 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% identity with at least one of the amino acid sequences shown in SEQ ID NO: 4-6. In some embodiments, the antibody binding to MICA / B comprises a heavy chain CDR sequence having an amino acid sequence having 100% identity with at least one of the amino acid sequences shown in SEQ ID NO: 4-6.

[0198] This document also discloses antibodies that bind to MICA / B, comprising a light chain complementarity-determining region (CDR) and a heavy chain complementarity-determining region (CDR). In some embodiments, the antibody that binds to MICA / B comprises a light chain CDR sequence and a heavy chain CDR sequence, wherein the light chain CDR sequence has an amino acid sequence having at least about 70% identity with at least one of the amino acid sequences shown in SEQ ID NO: 1-3, and the heavy chain CDR sequence has an amino acid sequence having at least about 70% identity with at least one of the amino acid sequences shown in SEQ ID NO: 4-6. In some embodiments, the antibody binding to MICA / B comprises a light chain CDR sequence and a heavy chain CDR sequence, wherein the light chain CDR sequence has an amino acid 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% identity with at least one of the amino acid sequences shown in SEQ ID NO: 1-3, and the heavy chain CDR sequence has an amino acid 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% identity with at least one of the amino acid sequences shown in SEQ ID NO: 4-6. In some embodiments, the antibody binding to MICA / B comprises a light chain CDR sequence and a heavy chain CDR sequence, wherein the light chain CDR sequence has an amino acid sequence that is 100% identical to at least one of the amino acid sequences shown in SEQ ID NO: 1-3, and the heavy chain CDR sequence has an amino acid sequence that is at least about 100% identical to at least one of the amino acid sequences shown in SEQ ID NO: 4-6.

[0199] In some embodiments, the antibody binding to MICA / B comprises at least one of light chain CDR1, light chain CDR2, and light chain CDR3, wherein light chain CDR1 has an amino acid sequence that is at least about 70% identical to the amino acid sequence shown in SEQ ID NO: 1, light chain CDR2 has an amino acid sequence that is at least about 70% identical to the amino acid sequence shown in SEQ ID NO: 2, and light chain CDR3 has an amino acid sequence that is at least about 70% identical to the amino acid sequence shown in SEQ ID NO: 3. In some embodiments, the antibody binding to MICA / B comprises at least one of light chain CDR1, light chain CDR2, and light chain CDR3, wherein light chain CDR1 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 shown in SEQ ID NO: 1; light chain CDR2 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 shown in SEQ ID NO: 2; and light chain CDR3 ... or 99% identical to the amino acid sequence shown in SEQ ID NO: 2; and light chain CDR3 has an amino acid sequence that is at least about 75%, 80%, 81%, 82%, 83%, 84%, 85%, 96 The amino acid sequence shown in SEQ ID NO: 1 has 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% amino acid sequence identity. In some embodiments, the antibody binding to MICA / B comprises at least one of light chain CDR1, light chain CDR2, and light chain CDR3, wherein light chain CDR1 has an amino acid sequence that is 100% identical to the amino acid sequence shown in SEQ ID NO: 1, light chain CDR2 has an amino acid sequence that is 100% identical to the amino acid sequence shown in SEQ ID NO: 2, and light chain CDR3 has an amino acid sequence that is 100% identical to the amino acid sequence shown in SEQ ID NO: 3.

[0200] In some embodiments, the antibody binding to MICA / B comprises at least one of heavy chain CDR1, heavy chain CDR2, and heavy chain CDR3, wherein heavy chain CDR1 has an amino acid sequence that is at least about 70% identical to the amino acid sequence shown in SEQ ID NO: 4, heavy chain CDR2 has an amino acid sequence that is at least about 70% identical to the amino acid sequence shown in SEQ ID NO: 5, and heavy chain CDR3 has an amino acid sequence that is at least about 70% identical to the amino acid sequence shown in SEQ ID NO: 6. In some embodiments, the antibody binding to MICA / B comprises at least one of heavy chain CDR1, heavy chain CDR2, and heavy chain CDR3, wherein heavy chain CDR1 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 shown in SEQ ID NO: 4; heavy chain CDR2 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 shown in SEQ ID NO: 5; and heavy chain CDR3 ... The amino acid sequence shown in SEQ ID NO: 4 has an amino acid sequence with 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% identity. In some embodiments, the antibody binding to MICA / B comprises at least one of heavy chain CDR1, heavy chain CDR2, and heavy chain CDR3, wherein heavy chain CDR1 has an amino acid sequence with 100% identity to the amino acid sequence shown in SEQ ID NO: 4, heavy chain CDR2 has an amino acid sequence with 100% identity to the amino acid sequence shown in SEQ ID NO: 5, and heavy chain CDR3 has an amino acid sequence with 100% identity to the amino acid sequence shown in SEQ ID NO: 6.

[0201] In some embodiments, the antibody binding to MICA / B comprises at least one of light chain CDR1, light chain CDR2, light chain CDR3, heavy chain CDR1, heavy chain CDR2, and heavy chain CDR3, wherein light chain CDR1 has an amino acid sequence having at least about 70% identity with the amino acid sequence shown in SEQ ID NO: 1, light chain CDR2 has an amino acid sequence having at least about 70% identity with the amino acid sequence shown in SEQ ID NO: 2, light chain CDR3 has an amino acid sequence having at least about 70% identity with the amino acid sequence shown in SEQ ID NO: 3, heavy chain CDR1 has an amino acid sequence having at least about 70% identity with the amino acid sequence shown in SEQ ID NO: 4, heavy chain CDR2 has an amino acid sequence having at least about 70% identity with the amino acid sequence shown in SEQ ID NO: 5, and heavy chain CDR3 has an amino acid sequence having at least about 70% identity with the amino acid sequence shown in SEQ ID NO: 6.In some embodiments, the antibody binding to MICA / B comprises at least one of light chain CDR1, light chain CDR2, light chain CDR3, heavy chain CDR1, heavy chain CDR2, and heavy chain CDR3, wherein light chain CDR1 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 shown in SEQ ID NO: 1; light chain CDR2 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 shown in SEQ ID NO: 2; and light chain CDR3 ... The amino acid sequence shown in SEQ ID NO: 3 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 shown in SEQ ID NO: 4. The heavy chain CDR1 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 shown in SEQ ID NO: 4. The amino acid sequence shown in SEQ ID NO: 6 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 shown in SEQ ID NO: 6.In some embodiments, the antibody binding to MICA / B comprises at least one of light chain CDR1, light chain CDR2, light chain CDR3, heavy chain CDR1, heavy chain CDR2, and heavy chain CDR3, wherein light chain CDR1 has an amino acid sequence that is 100% identical to the amino acid sequence shown in SEQ ID NO: 1, light chain CDR2 has an amino acid sequence that is 100% identical to the amino acid sequence shown in SEQ ID NO: 2, light chain CDR3 has an amino acid sequence that is 100% identical to the amino acid sequence shown in SEQ ID NO: 3, heavy chain CDR1 has an amino acid sequence that is 100% identical to the amino acid sequence shown in SEQ ID NO: 4, heavy chain CDR2 has an amino acid sequence that is 100% identical to the amino acid sequence shown in SEQ ID NO: 5, and heavy chain CDR3 has an amino acid sequence that is 100% identical to the amino acid sequence shown in SEQ ID NO: 6.

[0202] MICA / B antibody variable domain

[0203] This document discloses antibodies that specifically bind to MICA / B, said antibodies having a light chain comprising a light chain variable domain (VL). In some embodiments, the antibody binding to MICA / B comprises a light chain variable domain (VL) having an amino acid sequence having at least about 70% identity with the amino acid sequence shown in SEQ ID NO: 7. In some embodiments, the VL has an amino acid 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% identity with the amino acid sequence shown in SEQ ID NO: 7. In some embodiments, the VL has an amino acid sequence having 100% identity with the amino acid sequence shown in SEQ ID NO: 7.

[0204] This document further discloses antibodies that specifically bind to MICA / B, said antibodies having a heavy chain comprising a heavy chain variable domain (VH). In some embodiments, the antibody binding to MICA / B comprises a heavy chain variable domain (VH) having an amino acid sequence having at least about 70% identity with the amino acid sequence shown in SEQ ID NO: 8. In some embodiments, the VH has an amino acid 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% identity with the amino acid sequence shown in SEQ ID NO: 8. In some embodiments, the VH has an amino acid sequence having 100% identity with the amino acid sequence shown in SEQ ID NO: 8.

[0205] This document also discloses antibodies that bind to MICA / B, said antibodies comprising a light chain variable domain (VL) and a heavy chain variable domain (VH). In some embodiments, the antibody binding to MICA / B comprises a light chain variable domain (VL) and a heavy chain variable domain (VH), said VL having an amino acid sequence having at least about 70% identity with the amino acid sequence shown in SEQ ID NO: 7, and said VH having an amino acid sequence having at least about 70% identity with the amino acid sequence shown 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 shown 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 shown in SEQ ID NO: 8. In some embodiments, VL has an amino acid sequence that is 100% identical to the amino acid sequence shown in SEQ ID NO: 7, and VH has an amino acid sequence that is 100% identical to the amino acid sequence shown in SEQ ID NO: 8.

[0206] MICA / B antibody heavy and light chains

[0207] This document discloses antibodies that specifically bind to MICA / B, said antibodies having a light chain. In some embodiments, the antibody binding to MICA / B comprises a light chain having an amino acid sequence having at least about 70% identity with the amino acid sequence shown in SEQ ID NO: 9. In some embodiments, the light chain has an amino acid 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% identity with the amino acid sequence shown in SEQ ID NO: 9. In some embodiments, the light chain has an amino acid sequence having 100% identity with the amino acid sequence shown in SEQ ID NO: 9.

[0208] This document further discloses antibodies that specifically bind to MICA / B, said antibodies having a heavy chain. In some embodiments, the antibody binding to MICA / B comprises a heavy chain having an amino acid sequence having at least about 70% identity with the amino acid sequence shown in SEQ ID NO: 10. In some embodiments, the heavy chain has an amino acid 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% identity with the amino acid sequence shown in SEQ ID NO: 10. In some embodiments, the heavy chain has an amino acid sequence having 100% identity with the amino acid sequence shown in SEQ ID NO: 10.

[0209] This document also discloses antibodies that bind to MICA / B, said antibodies comprising a light chain and a heavy chain. In some embodiments, the antibody that binds to MICA / B comprises a light chain and a heavy chain, said light chain having an amino acid sequence having at least about 70% identity with the amino acid sequence shown in SEQ ID NO: 9, and said heavy chain having an amino acid sequence having at least about 70% identity with the amino acid sequence shown 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 shown in SEQ ID NO: 10, 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 shown 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 shown in SEQ ID NO: 9, and the heavy chain has an amino acid sequence that is 100% identical to the amino acid sequence shown in SEQ ID NO: 10.

[0210] Treatment methods and uses

[0211] This article provides a method for treating cancer in individuals in need, which includes administering the 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 and soluble MICB proteins. In some embodiments, the antibodies disclosed herein reduce the shedding of soluble MICA protein. In some embodiments, the antibodies disclosed herein reduce the shedding of soluble MICB protein. In some embodiments, the antibodies disclosed herein reduce the shedding of both soluble MICA and soluble MICB proteins. 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 and soluble MICB proteins. In some embodiments, treatment using the disclosed anti-MICA / B antibody alleviates or suppresses the immunosuppressive environment by preventing or blocking the interaction between soluble MICA / B and NKG2D receptors in an individual.

[0213] Subjects

[0214] In some embodiments, the subjects described herein are subjects with cancer. Subjects may be mammals, such as primates, higher primates, such as humans (e.g., patients with the condition described herein, such as cancer, or at risk of developing the condition described herein, such as cancer). In one embodiment, the subject needs to enhance an immune response. In one embodiment, the subject has the condition described herein, such as cancer, or is at risk of developing the condition described herein, such as cancer. In some embodiments, the subject has weakened immune function or is at risk of weakened immune function.

[0215] In some implementations, the subject to be treated is a subject who has previously been treated with checkpoint inhibitor therapy. In some implementations, the checkpoint inhibitor therapy includes a PD-1 inhibitor (e.g., an anti-PD-1 antibody). In some implementations, the subject responds to the PD-1 inhibitor (e.g., with a complete or partial response). In some implementations, 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 after administration of the PD-1 inhibitor. In some implementations, the subject's disease progresses after the response period to the PD-1 inhibitor. In some implementations, after disease progression following the response period to the PD-1 inhibitor, the subject receives treatment with an anti-MICA / B antibody.

[0216] In some implementations, the subject has stable disease after administration of a PD-1 inhibitor. In some implementations, the subject has a stable disease 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. In some implementations, the subject's disease progresses after a stable disease period following PD-1 inhibitor treatment. In some implementations, after disease progression following a stable disease period following PD-1 inhibitor treatment, the subject receives treatment with an anti-MICA / B antibody.

[0217] In some implementations, PD-1 inhibitor therapy is discontinued before the initiation of anti-MICA / B antibody treatment. In some implementations, PD-1 inhibitor therapy is continued after the initiation of anti-MICA / B antibody treatment.

[0218] In some implementations, treatment with anti-MICA / B antibodies elicits a response (e.g., a complete or partial response) or stabilizes the disease. In some implementations, the response following 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, 60 months or longer.

[0219] In some implementations, PD-1 inhibitors include pimiralimumab, pembrolizumab, nivolumab, cimiprimab, AMP-224, APM-514, or spartazolizumab.

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

[0221] In some implementations, the subject to be treated is a subject who has previously been treated with a VEGF inhibitor. In some implementations, the subject responds to the VEGF inhibitor (e.g., with a complete or partial response). In some implementations, 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 after administration of the VEGF inhibitor. In some implementations, the subject's disease progresses after the response period to the VEGF inhibitor. In some implementations, after disease progression following the response period to the VEGF inhibitor, the subject receives treatment with an anti-MICA / B antibody.

[0222] In some implementations, the subject has stable disease after administration of a VEGF inhibitor. In some implementations, the subject has a stable disease 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. In some implementations, the subject's disease progresses after a stable disease period following treatment with a VEGF inhibitor. In some implementations, after disease progression following a stable disease period following treatment with a VEGF inhibitor, the subject receives treatment with an anti-MICA / B antibody.

[0223] In some implementations, VEGF inhibitor therapy is discontinued before the initiation of anti-MICA / B antibody treatment. In some implementations, VEGF inhibitor therapy is continued after the initiation of anti-MICA / B antibody treatment.

[0224] In some implementations, treatment with anti-MICA / B antibodies elicits a response (e.g., a complete or partial response) or stabilizes the disease. In some implementations, the response following 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, 60 months or longer.

[0225] In some implementations, VEGF inhibitors include small molecule tyrosine kinase inhibitors (TKIs) (e.g., sunitinib, sorafenib, axitinib, pazopanib, or lenvatinib) or anti-VEGF antibodies (e.g., bevacizumab or ranibizumab).

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

[0227] In some implementations, the subject to be treated is a subject who has previously been treated with an EGFR inhibitor. In some implementations, the subject responds to the EGFR inhibitor (e.g., with a complete or partial response). In some implementations, 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 after administration of the EGFR inhibitor. In some implementations, the subject's disease progresses after the response period to the EGFR inhibitor. In some implementations, after disease progression following the response period to the EGFR inhibitor, the subject receives treatment with an anti-MICA / B antibody.

[0228] In some implementations, the subject has stable disease after administration of an EGFR inhibitor. In some implementations, the subject has a stable disease 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. In some implementations, the subject's disease progresses after a stable disease period following EGFR inhibitor treatment. In some implementations, the subject receives treatment with an anti-MICA / B antibody after disease progression following a stable disease period following EGFR inhibitor treatment.

[0229] In some implementations, EGFR inhibitor therapy is discontinued before the initiation of anti-MICA / B antibody treatment. In other implementations, EGFR inhibitor therapy is continued after the initiation of anti-MICA / B antibody treatment.

[0230] In some implementations, treatment with anti-MICA / B antibodies elicits a response (e.g., a complete or partial response) or stabilizes the disease. In some implementations, the response following 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, 60 months or longer.

[0231] In some implementations, EGFR inhibitors include small molecule tyrosine kinase inhibitors (TKIs) (e.g., gefitinib, erlotinib, afatinib, dacomitinib, or osimertinib) or anti-EGFR antibodies (e.g., cetuximab, panitumumab, or nexituzumab).

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

[0233] In some implementations, the subject to be treated is a subject who has previously been treated with hormone therapy. In some implementations, the subject responds to hormone therapy (e.g., with a complete or partial response). In some implementations, 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 after hormone therapy administration. In some implementations, the subject's disease progresses after the response period to hormone therapy. In some implementations, after disease progression following the response period to hormone therapy, the subject receives treatment with an anti-MICA / B antibody.

[0234] In some implementations, the subject has stable disease after administration of hormone therapy. In some implementations, the subject has a stable disease 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. In some implementations, the subject's disease progresses after a stable disease period following hormone therapy. In some implementations, after disease progression following a stable disease period following hormone therapy, the subject receives treatment with an anti-MICA / B antibody.

[0235] In some implementations, hormone therapy is discontinued before the administration of anti-MICA / B antibodies is started. In some implementations, hormone therapy is continued after the administration of anti-MICA / B antibodies is started.

[0236] In some implementations, treatment with anti-MICA / B antibodies elicits a response (e.g., a complete or partial response) or stabilizes the disease. In some implementations, the response following 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, 60 months or longer.

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

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

[0239] In some embodiments, the subject to be 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 one or both of ER+ or PR+. In some embodiments, the cancer is mutated for one or more of the following: EGFR, TP53, HER2, PTEN, ARID1A, CTNNB1, or PLK3R1.

[0240] In some implementations, the subject to be treated is a subject with parotid gland cancer. In some implementations, the parotid gland cancer overexpresses one or both of EGFR and HER2. In some implementations, the parotid gland cancer is mucoepidermoid parotid carcinoma. In some implementations, the cancer is mutated for one or more of the following: EGFR, TP53, HER2, PTEN, ARID1A, CTNNB1, or PLK3R1.

[0241] cancer

[0242] In some embodiments, the cancers treated according to the methods described herein include, but are not limited to, female reproductive tract tumors, 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, the composition (including, but not limited to, pharmaceutical compositions containing anti-MICA / B antibodies) is particularly useful for treating high (large) tumor burden. Tumor burden refers to the number of cancer cells, the size of the tumor, or the amount of cancer in the body. High tumor burden can be used as a clinical biomarker with negative prognostic value in a variety of solid tumors.

[0244] In some embodiments, pharmaceutical compositions comprising anti-MICA / B antibodies are particularly suitable for treating female reproductive tract tumors. In some embodiments, the female reproductive tract tumors are endometrial tumors, ovarian cancer, or cervical cancer. In some embodiments, salivary gland tumors are mucoepidermoid tumors, such as parotid gland tumors or adenoid cystic sarcoma. In some embodiments, the cancer is virally caused cancer. Virally caused cancers include cancers associated with human papillomavirus (HPV), Epstein-Barr virus (EBV), or cytomegalovirus (CMV) infection. In some embodiments, the 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, the cancer is urothelial carcinoma of the bladder. In some embodiments, the cancer is adrenocortical carcinoma. In some embodiments, the cancer is squamous cell carcinoma of the head and neck. In some embodiments, cervical cancer is squamous cell carcinoma of the cervix or intracervical carcinoma. In some embodiments, the cancer is esophageal cancer. In some embodiments, the cancer is squamous cell carcinoma of the lung. In some embodiments, the cancer is diffuse large B-cell lymphoma. In some embodiments, the cancer is thymoma. In some embodiments, the cancer is acute myeloid leukemia (AML). In some embodiments, the cancer is uveal melanoma. In some embodiments, the cancer is prostate adenocarcinoma. In some embodiments, the cancer is chromophobe renal cell carcinoma. In some embodiments, the cancer is low-grade glioma of the brain. In some embodiments, the cancer is hepatocellular carcinoma of the liver. In some embodiments, the cancer is cutaneous melanoma of the skin. In some embodiments, the cancer is pheochromocytoma or paraganglioma. In some embodiments, the cancer is carcinosarcoma of the uterus. In some embodiments, the cancer is ovarian serous cystadenocarcinoma. In some embodiments, the cancer is 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 clear cell renal 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 of the uterine corpus. In some embodiments, the cancer is rectal adenocarcinoma. In some embodiments, the cancer is colonic adenocarcinoma.

[0246] In some implementations, the cancer has metastasized. In some implementations, the cancer is mutated for one or more of the following: 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 burden (TMB). In some embodiments, the cancer is characterized by high tumor mutational burden (TMB). In some embodiments, the cancer is an immune-cold cancer. In some embodiments, the cancer is an immune-hot cancer. In some embodiments, the cancer is a hormone-sensitive cancer. In some embodiments, the cancer is characterized by overexpression of oncogenic drivers. In some embodiments, the cancer expresses one or more of the following: EGFR, ER, PR, or HER2.

[0248] Dosage and Dosing Regimen

[0249] The contemplated therapeutically effective doses or dosages include the following: 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 1.2 mg / kg, approximately 1.3 mg / kg, approximately 1.4 mg / kg, approximately 1.5 mg / kg, approximately 1.6 mg / kg, approximately 1.7 mg / kg, etc. mg / kg, approximately 1.8 mg / kg, approximately 1.9 mg / kg, approximately 2 mg / kg, approximately 2.1 mg / kg, approximately 2.2 mg / kg, approximately 2.3 mg / kg, approximately 2.4 mg / kg, approximately 2.5 mg / kg, approximately 2.6 mg / kg, approximately 2.7 mg / kg, approximately 2.8 mg / kg, approximately 2.9 mg / kg, approximately 3 mg / kg, approximately 3.1 mg / kg, approximately 3.2 mg / kg, approximately 3.3 mg / kg, approximately 3.4 mg / kg, approximately 3.5 mg / kg, approximately 3.6 mg / kg, approximately 3.7 mg / kg, approximately 3.8 mg / kg, approximately 3.9 mg / kg, approximately 4 mg / kg, approximately 4.1 mg / kg, approximately 4.2 mg / kg, approximately 4.3 mg / kg, approximately 4.4 mg / kg, approximately 4.5 mg / kg, approximately 4.6 mg / kg, approximately 4.7 mg / kg, approximately 4.8 mg / kg. mg / kg, approximately 4.9 mg / kg, approximately 5 mg / kg, approximately 5.1 mg / kg, approximately 5.2 mg / kg, approximately 5.3 mg / kg, approximately 5.4 mg / kg, approximately 5.5 mg / kg, approximately 5.6 mg / kg, approximately 5.7 mg / kg, approximately 5.8 mg / kg, approximately 5.9 mg / kg, approximately 6 mg / kg, approximately 6.1 mg / kg, approximately 6.2 mg / kg, approximately 6.3 mg / kg, approximately 6.4 mg / kg, approximately 6.5 mg / kg, approximately 6.6 mg / kg, approximately 6.7 mg / kg, approximately 6.8 mg / kg, approximately 6.9 mg / kg, approximately 7 mg / kg, approximately 7.1 mg / kg, approximately 7.2 mg / kg, approximately 7.3 mg / kg, approximately 7.4 mg / kg, approximately 7.5 mg / kg, approximately 7.6 mg / kg, approximately 7.7 mg / kg, approximately 7.8 mg / kg, approximately 7.9 mg / kg, approximately 8 mg / kg, approximately 8.1 mg / kg, approximately 8.2 mg / kg, approximately 8.3 mg / kg, approximately 8.4 mg / kg, approximately 8.5 mg / kg, approximately 8.6 mg / kg, approximately 8.7 mg / kg, approximately 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. mg / kg, approximately 11 mg / kg, approximately 11.1 mg / kg, approximately 11.2 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. 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 15.8 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 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 18.1 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. 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 about 0.1 mg / kg to about 10 mg / kg. In some embodiments, the anti-MICA / B antibody is administered at a dose of about 3 mg / kg to about 10 mg / kg. In some embodiments, the anti-MICA / B antibody is administered at a dose of about 0.1 mg / kg. In some embodiments, the anti-MICA / B antibody is administered at a dose of about 0.3 mg / kg. In some embodiments, the anti-MICA / B antibody is administered at a dose of about 1 mg / kg. In some embodiments, the anti-MICA / B antibody is administered at a dose of about 3 mg / kg. In some embodiments, the anti-MICA / B antibody is administered at a dose of about 6 mg / kg. In some embodiments, the anti-MICA / B antibody is administered at a dose of about 10 mg / kg.

[0251] The treatment methods described herein include administering anti-MICA / B antibodies once or multiple times at the doses disclosed herein. In some embodiments, the method includes administering anti-MICA / B antibodies once. In some embodiments, the method includes administering anti-MICA / B antibodies twice. In some embodiments, the method includes administering anti-MICA / B antibodies three times. In some embodiments, the method includes administering anti-MICA / B antibodies four times. In some embodiments, the method includes administering anti-MICA / B antibodies five times. In some embodiments, the method includes administering anti-MICA / B antibodies six times. In some embodiments, the method includes administering anti-MICA / B antibodies more than six times.

[0252] In some embodiments, the anti-MICA / B antibody is administered according to a dosing interval (e.g., a cycle). In some embodiments, the anti-MICA / B antibody is administered once or more daily. In some embodiments, the anti-MICA / B antibody is administered once or more weekly. In some embodiments, the anti-MICA / B antibody is administered once or more every two weeks (Q2W). In some embodiments, the anti-MICA / B antibody is administered once or more every three weeks (Q3W). In some embodiments, the anti-MICA / B antibody is administered once or more monthly. In some embodiments, the anti-MICA / B antibody is administered once or more every three months. In some embodiments, the anti-MICA / B antibody is administered once or more every six months. In some embodiments, the anti-MICA / B antibody is administered once or more 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 comprises a three-week cycle, and the anti-MICA / B antibody is administered once every three 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 repeated dosing interval is performed 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.

[0254] In some implementations, subjects receive one or more doses as a prodrug before administration of the anti-MICA / B antibody.

[0255] In some implementations, the prophylactic drug includes acetaminophen or ibuprofen. In some implementations, the subject receives acetaminophen or ibuprofen 30-60 minutes prior to administration of the anti-MICA / B antibody. In some implementations, the prophylactic drug is administered orally.

[0256] In some embodiments, the prodrug includes a corticosteroid. In some embodiments, the subject receives the corticosteroid 30-60 minutes before administration of the anti-MICA / B antibody. In some embodiments, the corticosteroid is administered before the first 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 at a dose of about 2 mg to about 50 mg, or virtually any standard-of-care corticosteroid dose used in cancer therapy. In some embodiments, the corticosteroid includes dexamethasone. As in Example 1 and... Figure 5 As shown, an increase in cytokine levels was observed in patients after the first dose of anti-MICA / B antibody, but subsequent doses of anti-MICA / B antibody caused a smaller increase or no increase. It is not desirable to be bound by theory that administration of corticosteroids prior to the first administration of anti-MICA / B antibody could suppress the patient's immune response to the administered antibody.

[0257] Any suitable route of administration is contemplated for use with the methods disclosed herein. In some embodiments, the antibody is administered intravenously. In some embodiments, the antibody is administered subcutaneously. In some embodiments, the antibody is administered orally. In some embodiments, the antibody is administered systemically (e.g., intravenously, intramuscularly, subcutaneously, intradermally, 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 solution. In some embodiments, the antibody is formulated for topical, oral, buccal, or nasal administration.

[0258] In some implementations, the anti-MICA / B antibody is administered intravenously over approximately 1 hour.

[0259] In some embodiments, individuals are monitored prior to antibody administration. Symptoms are identified and their severity is assessed. Antibodies, as described herein, may be administered alone or in combination with other treatments, administered once or multiple times over time, as described herein or known to those skilled in the art. In some embodiments, individuals are monitored to determine the efficacy of the treatment regimen. In some embodiments, the treatment regimen is modified in response to preliminary treatment results, such as changing the treatment dose or frequency, or dose and frequency, to achieve the desired level of subject response in terms of symptom relief, reduction of side effects, or a combination of symptom relief and reduction of side effects.

[0260] In some embodiments, the treatments disclosed herein are monotherapy. In some embodiments, the treatments disclosed herein are combination therapies. In some embodiments, combination therapies include administration of an anti-MICA / B antibody in combination with another therapeutic agent. In this regard, when treating immunosuppressive TMEs in a clinical setting, the full killing potential of NK cells may require more than one activating stimulus, such as the simultaneous involvement of CD16A and NKG2D. The ultimate therapeutic potential of NK cell activating agents in clinical practice may be realized when delivered in combination with therapies that enhance or supplement NK cell activation, such as lenalidomide, cytokines, or checkpoint inhibitors. Combinations with checkpoint inhibitors provide the opportunity for dual activation of innate and adaptive immune responses and may alleviate the depletion of PD-1-expressing T cells and NK cells. Therefore, this disclosure envisions the following immuno-oncology therapies in which anti-MICA / B antibodies can be administered in combination with certain inhibitors of immune checkpoint molecules, namely programmed cell death 1 receptor (PD-1), T cell immune receptor with Ig and ITIM domains (TIGIT), protein 3 containing T cell immunoglobulin and mucin domains (TIM-3), differentiation cluster 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 is considered a prototypical immune checkpoint molecule that negatively regulates T cell function on cancer cells and immune cells in the tumor microenvironment (TME) by interacting with programmed death ligands 1 and 2 (PD-L1 and PD-L2), and this axis has been successfully targeted clinically with antagonist mAb therapy. Checkpoint inhibitors can be administered before, after, or concurrently with the anti-MICA / B therapy of this disclosure. Tumor-acquired NK cells have also been shown to express PD-1 and PD-L1 in preclinical and clinical settings, so using a combination of PD-1 inhibition with MICA / B antibodies is a specific implementation scheme envisioned in this paper.

[0261] In some embodiments, the composition of this disclosure comprising an anti-MICA / B antibody is in a pharmaceutical combination or therapy together with at least one additional therapeutic agent selected from the family of: antibodies, antibody fragments, antibody conjugates, cytotoxic agents, toxins, radionuclides, immunomodulators, photoactive therapeutic agents, radiosensitizers, hormones, anti-angiogenic agents, and combinations thereof.

[0262] In some embodiments, the therapeutic agent includes chemotherapeutic agents. 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, anthracene diones, anthracyclines, epipodophyllotoxin, quinoline alkaloids, etc.), antimicrotubule agents (e.g., taxanes, vinca alkaloids), protein synthesis inhibitors (e.g., holothurin, camptothecin derivatives, quinoline alkaloids), alkylating agents (e.g., alkyl sulfonates, ethyleneimine, nitrogen mustard, nitrosourea, platinum derivatives, triazine, etc.), alkaloids, terpenoids, kinase inhibitors, and immune checkpoint inhibitors.

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

[0264] In some embodiments, the anti-MICA / B antibody disclosed herein is administered in combination with a therapeutic agent that induces an immune response. In some embodiments, the anti-MICA / B antibody disclosed herein is administered in combination with a therapeutic agent that downregulates an immune response. In some embodiments, inducing an immune response includes activating or upregulating NK cell activity. In some embodiments, inducing an immune response includes activating or upregulating 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] Specific, non-limiting examples of immune checkpoint inhibitors targeting the PD-1 / PD-L1 pathway include, but are not limited to, pimiralimumab, pembrolizumab (Keytruda, MK-3475, lambolizumab), nivolumab (e.g., Opdivo, BMS-936558, MDX1106), cimiprimab, spartazolizumab (PDR001), STI-A1110, AMP-224, AMP-514 (MEDI0680), JS001, atezolizumab (e.g., Tecentriq, MPDL3280A), avelumab (MSB0010718C), durvalumab (MEDI4736, BMS-936559), and pidilizumab (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 comprises a heavy chain and a light chain, said heavy chain comprising the following amino acid sequence: QVQLVQSGVEVKKPGASVKVSCKASGYTFTNYYMYWVRQAPGQGLEWMGGINPSNGGTNFNEKFKNRVTLTTDSSTTTAYMELKSLQFDDTAVYYCARRDYRFDMGFDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVD HKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK GLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 11), and the light chain comprises the following 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 even PD-L1 inhibitors may be used in place 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 every three weeks (Q3W). In some embodiments, the subject receiving combination therapy including an immune checkpoint inhibitor (e.g., pembrolizumab) has ovarian cancer, prostate cancer, rectal cancer, NSCLC, squamous cell carcinoma of the skin, 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 molecule 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 molecule tyrosine kinase inhibitor (TKI) (e.g., gefitinib, erlotinib, afatinib, dacomitinib, or osimertinib) or an anti-EGFR antibody (e.g., cetuximab, panitumumab, or nexituzumab). In some embodiments, the anti-MICA / B antibody described herein is administered in combination with hormone therapy. In some embodiments, the hormone therapy includes an aromatase inhibitor (AI) (e.g., anastrozole), a selective estrogen receptor modulator (SERM), a luteinizing hormone-releasing hormone (LHRH) agonist, an anti-androgen, a CYP17 inhibitor, a progestin, an anti-adrenergic drug, or an estrogen receptor antagonist. In some embodiments, the anti-MICA / B antibody described herein is administered in combination with radiation therapy. In some embodiments, the anti-MICA / B antibody described herein is administered in combination with one or more cell therapies.

[0268] Exemplary chemical structures of some of the therapeutic agents described herein are shown in Table 2.

[0269] Table 2: Exemplary chemical structures of therapeutic agents

[0270]

[0271]

[0272]

[0273]

[0274] Pharmaceutical Composition

[0275] This article also discloses pharmaceutical compositions comprising the anti-MICA / B antibody disclosed herein and a pharmaceutically acceptable carrier or excipient.

[0276] In some embodiments, excipients used 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 surfactant polyoxyethylene-sorbitan monooleate.

[0277] In some embodiments, the composition further comprises additional therapeutic agents. In some embodiments, the therapeutic agent is a chemotherapeutic agent. Chemotherapy agents may include, in particular, cytotoxic agents, antimetabolites (e.g., folic acid antagonists, purine analogs, pyrimidine analogs, etc.), topoisomerase inhibitors (e.g., camptothecin derivatives, anthracene diones, anthracyclines, epipodophyllotoxin, quinoline alkaloids, etc.), antimicrotubule agents (e.g., taxanes, vinca alkaloids), protein synthesis inhibitors (e.g., holothurin, camptothecin derivatives, quinoline alkaloids), alkylating agents (e.g., alkyl sulfonates, ethyleneimine, nitrogen mustard, nitrosourea, platinum derivatives, triazine, etc.), alkaloids, terpenoids, and kinase inhibitors.

[0278] In some embodiments, the antibody and the therapeutic agent are in the same formulation. In some embodiments, the antibody and the therapeutic agent are 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 concurrently with the administration of other therapeutic agents. In some embodiments, the antibody described herein is used after the administration of other therapeutic agents.

[0279] In some embodiments, the pharmaceutical formulation is formulated to be compatible with a specific local, regional, or systemic route of administration or delivery. Therefore, the pharmaceutical formulation contains a carrier, diluent, or excipient suitable for administration via a specific route. Specific, non-limiting examples of routes of administration for the compositions herein are parenteral administration, such as intravenous, intra-arterial, intradermal, intramuscular, subcutaneous, intrapleural, percutaneous (local), transmucosal, intracranial, intraspinal, intraocular, rectal, oral (gastrointestinal), mucosal administration, and any other formulation suitable for a treatment method or administration regimen.

[0280] In some embodiments, the solution or suspension for parenteral application comprises: a sterile diluent, such as water for injection, saline solution, non-volatile oil, polyethylene glycol, glycerin, propylene glycol, or other synthetic solvent; an antibacterial agent, such as benzyl alcohol or methylparaben; an antioxidant, such as ascorbic acid or sodium bisulfate; a chelating agent, such as ethylenediaminetetraacetic acid; a buffer, such as acetate, citrate, or phosphate; and an agent for adjusting tension, such as sodium chloride or dextrose. In some embodiments, the pH is adjusted with an acid or base, such as hydrochloric acid or sodium hydroxide.

[0281] Pharmaceutical formulations for injection include sterile aqueous solutions (in the case of water solubility) or dispersions and sterile powders for the ad hoc preparation of sterile injectable solutions or dispersions. For intravenous administration, suitable carriers include physiological saline, antibacterial 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. In some embodiments, flowability is maintained, for example, by using coatings (such as lecithin), by maintaining the desired particle size (in the case of dispersions), and by using surfactants. Antibacterial and antifungal agents include, for example, parabens, chlorobutanol, phenol, ascorbic acid, and thimerosal. In some embodiments, the composition contains isotonic agents, such as sugars; polyols, such as mannitol or sorbitol; or sodium chloride. In some cases, an absorption-delaying agent is also included; in some embodiments, aluminum monostearate or gelatin prolongs the absorption of the injectable composition.

[0282] In some embodiments, sterile injectable formulations are prepared by incorporating a desired amount of the active composition with one or a combination of the aforementioned ingredients into a suitable solvent. Typically, dispersions are prepared by incorporating the active composition into a sterile medium containing an alkaline dispersion medium and any other ingredients. In the case of sterile powders used to prepare sterile injectable solutions, preparation methods include, for example, vacuum drying and freeze-drying, methods that yield powders of the active ingredient and any other desired ingredients from previously prepared solutions.

[0283] For transmucosal or transdermal application, a penetrant suitable for the permeability barrier is used in the formulation. Such penetrants are known in the art and include, for example (for transmucosal application), cleansers, bile salts, and fusidic acid derivatives. In some embodiments, transmucosal application is achieved by using a nasal spray, an inhalation device (e.g., a suction device), or a suppository. For transdermal application, the active compound is formulated as an ointment, cream, gel, lotion, or patch.

[0284] In some embodiments, the pharmaceutical formulation is prepared together with a carrier that prevents rapid elimination from the body, such as a controlled-release formulation or a time-delayed material, such as glyceryl monostearate or glyceryl stearate. In some embodiments, articles such as implants and microcapsule delivery systems are also used to deliver the formulation to achieve local, regional, or systemic delivery or controlled or sustained release.

[0285] For NSCLC patients with relapsed after TKI therapy and targeted oncogenic mutations, a significant unmet need remains, thus we see the potential benefit of novel therapies that can be easily combined with established CPIs. Combining CLN-619 with pembrolizumab engages multiple immune effector cells, including innate cells (CLN-619) and T cells (pembrolizumab). The safety profile of CLN-619 and the biological basis for its combination with CPIs make it a potential synergistic approach.

[0286] Preliminary clinical results show that the combination of CLN-619 (a novel antibody targeting MICA / B) and pembrolizumab can benefit cancer patients who are typically ineligible for checkpoint inhibitor therapy. More specifically, the inventors observed an objective response in patients with relapsed ALK and EGFR-mutant NSCLC who typically do not respond to checkpoint inhibitors, following treatment with tyrosine kinase inhibitors (TKIs).

[0287] Furthermore, long-term follow-up of patients treated with CLN-619 monotherapy showed that extended treatment had a good safety profile and durable clinical benefits, including objective responses and long-term stable disease in patients with various tumor types and those whose disease progressed after CPI therapy.

[0288] Example

[0289] The following examples are provided to illustrate various embodiments of the invention, but are not intended to limit the invention in any way. These examples and the methods described herein represent certain embodiments and are exemplary, and are not intended to limit the scope of the invention. Variations and other uses will be apparent to those skilled in the art, and these variations and other uses are all covered within the spirit of the invention as defined by the claims.

[0290] Example 1. CLN-619 (anti-MICA / B antibody) promotes innate immune cell-mediated antitumor activity.

[0291] CLN-619 is a humanized IgG1 monoclonal antibody targeting MICA and MICB (MICA / B) and is currently in Phase 1 clinical development for cancer patients (NCT05117476; see examples below). This example demonstrates that the antibody promotes the antitumor activity of innate immune cells.

[0292] MICA / B acts as an activation signal on target cells, recognized by the NKG2D receptor, which is expressed on subsets of NK and T cell populations. In response to stress, MICA / B expression is induced, leading to NKG2D-mediated elimination of target cells. On NK cells, NKG2D is one of many receptors in a complex network of activating and inhibiting receptors, where activation of the NKG2D pathway leads to cytokine production, enhanced ADCC, and target cell death. On CD8 T cells, the NKG2D axis plays a co-stimulatory role in lowering the lysis threshold during TCR involvement and can also drive direct CD8-mediated killing following prior TCR activation. MICA / B is widely expressed in various solid tumors and hematologic malignancies. However, tumor cells evade NKG2D-mediated elimination by detaching MICA / B ligands from their cell surface via proteases present in the tumor microenvironment (TME). CLN-619 exerts its effects by binding to and preventing MICA / B cells from detaching, thereby increasing MICA / B cell surface expression to restore the NKG2D-MICA / B axis and promoting the killing of tumor cells by NK cells and T cells. Furthermore, CLN-619 can mediate ADCC and ADCP in NK cells and macrophages. In preclinical xenograft models, doses as low as 0.03 mg / kg were sufficient to inhibit tumor growth.

[0293] The inventors investigated the effects of CLN-619 on a variety of primary immune cells, including NK cells, T cells, and macrophages. In assessing NK cells, the regulation of cytokine production and cytotoxicity by CLN-619 was measured in the presence of MICA / B-expressing target cells. Furthermore, this embodiment demonstrates the contribution of Fc-mediated CLN-619 function. In the case of T cells, the necessity of TCR co-stimulation or prior antigen exposure for CLN-619 activation of the NKG2D pathway was explored. In the case of macrophages, the inventors measured the ability of CLN-619 to mediate ADCP against MICA / B-expressing target cells. These data highlight the potential of CLN-619 to engage multiple immune cell types within the TME, which may lead to greater and broader efficacy compared to IO therapies targeting a single immune cell type.

[0294] A. Method

[0295] CLN-619 is characterized by its binding epitope and affinity, its effects on the surface and solubility levels of MICA / B, and its in vitro tumor cell killing effect. In a mouse model, the tumor growth inhibition of the mAb was tested. The contribution of the functional Fc-gamma 1 (Fcγ1) domain to CLN-619 activity was also evaluated.

[0296] The creation of CLN-619

[0297] To generate multiple monoclonal antibodies against the human MICA / B protein, a repeated immunization multisite (RIMMS) 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]. Humanization of the primary candidate was performed using computer modeling.

[0298] Evaluation of CLN-619 in Octet Combination with MICA and MICB

[0299] To determine monovalent affinity, CLN-619 antibody (10 µg / ml) was captured on Dip and Readα anti-human IgG Fc capture biosensors. The sensors were 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 1:1 binding fitting algorithm (ForteBio analysis suite 8.0). The BLOSUM62 algorithm was used to determine the similarity of the MICA / B proteins.

[0300] ELISA assessment of the binding of CLN-619 to MICA / B allele variants

[0301] A 96-well plate was coated with monomerically his-labeled MICA / B as the capture antigen, and then serially diluted CLN-619 was added. Unbound material was washed away, and horseradish peroxidase (HRP)-conjugated mouse anti-human IgG Fc was added to the wells for detection.

[0302] Luminex evaluation of the combination of CLN-619 and MICA

[0303] The binding of CLN-619 (10 μg / mL) and the positive control antibody 6D4 to 28 different MICA allele variants was assessed using Luminex (LifeCodes LSA MIC Luminex kit) [Ghadially H et al. Br J Cancer. 2017; 116:1208-17]. 6D4 was detected using goat anti-mouse secondary antibody. Background MFI was subtracted from the original MFI, and the average of replicates was taken.

[0304] X-ray crystallography of MICA-CLN-619

[0305] X-ray diffraction data were collected from the crystal of Fab CLN-619:MICA using optimized low-temperature conditions at the Swiss Light Source (SLS, Villigen, Switzerland). The structure of the complex of the human Fab antibody fragment CLN-619 with the α3 domain of MICA (Fab CLN-619:MIC-A) was obtained at 2.12 Å resolution.

[0306] Flow cytometry assessment of surface MICA / B levels after CLN-619 treatment

[0307] 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 viability dyes. Cell surface MICA / B was detected using 6D4 antibody, a non-competitive anti-human MICA / B-PE antibody against CLN-619. Mouse IgG2a-PE isotype antibody was used as a control. Surface MICA / B levels were quantified using Quantibrite beads. A 4-parameter logistic regression model was used to fit dose-response curves.

[0308] Assessment of soluble MICA / B levels after CLN-619 treatment

[0309] HCC1534, PLC / PRF / 5, and HCT-116 were inoculated and treated with serially diluted CLN-619 or hIgG1 in duplicate for 24 hours. The supernatant was collected for analysis of sMICA levels in an enzyme-linked immunosorbent assay (ELISA). The antibody used in the ELISA was non-competitive with CLN-619. The ELISA plate was coated with BAMO1 capture antibody. A standard curve of recombinant human MICA*001 ECD and supernatant samples was added to the plate. Biotinylated detection antibody 10E9.H6 was added to the plate, followed by streptavidin-horseradish peroxidase (HRP). EC50 values ​​were calculated using a four-parameter nonlinear fitting model in GraphPad Prism.

[0310] Primary NK cell killing assay

[0311] In the xCELLigence system, the ability of CLN-619-mediated NK cell-mediated tumor cell killing was evaluated in a co-culture assay of purified primary human NK cells and MICA-expressing HCC1534 tumor cells. NK cells were isolated and their purity examined by negative selection from a stock of frozen PBMCs from healthy human donors. HCC1534 target cells were seeded onto E-96 Xcelligence plates containing serially titrated antibodies. After HCC1534 cell adhesion, NK cells were added at an effector cell to target cell ratio (E:T) of 40:1, and HCC1534 cell death was monitored over 72 hours. EC50 values ​​were calculated using a four-parameter nonlinear fitting model in GraphPad Prism.

[0312] ADCC Report Measurement

[0313] 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 the Activated T cell nuclear factor (NFAT)-luciferase reporter construct. HCT-116 or HCC1534 cells were treated with serially diluted CLN-619 or IgG1 control antibodies. Engineered Jurkat cells were added at an E:T ratio of 10:1 for 6 hours. EC50 values ​​were calculated using a four-parameter nonlinear fitting model in GraphPad Prism.

[0314] Primary cell ADCP assay

[0315] CD14+ monocytes were collected from two healthy PBMC donors using RoboSep (protocol 17858) or manual separation (StemCell) via magnetic positive selection. Macrophages were differentiated for 6 days, then harvested and seeded in M-CSF-supplemented medium. HCC1534 or HCT-116 eFluor670-labeled target cells and macrophage effector cells were seeded at an E:T ratio of 0.5:1 and incubated for 2 hours with the highest dose of serially diluted CLN-619 or hIgG1 or cetuximab control antibody. After incubation, macrophages were sequentially incubated with viability dye, blocking buffer (Biolegend), and BV510-labeled anti-CD64 antibody. ADCP was measured by flow cytometry by fluorescence gating of target cells within CD64+ cells (eF670+).

[0316] Flow cytometry assessment of MICA binding to NKG2D on NK cells.

[0317] NK cells were purified from PBMCs of healthy donors using negative selection. Serially diluted CLN-619, CLN-619DANA, or hIgG1 antibodies were pre-incubated with his-labeled MICA ECD at a ratio of 10:1. The antibody-MICA complex was then added to the NK cells with or without anti-NKG2D antibody. The binding of MICA to NK cells was detected by anti-His antibody.

[0318] Biacore assessment of CLN-619 and Fcγ receptor

[0319] To evaluate the binding of CLN-619 to the Fcγ receptor (FcγR), recombinant His6-labeled FcγR was immobilized onto a CM5 chip coated with an anti-His6 antibody. CLN-619 WT or CLN-619 DANA was then passed through the biosensor. Except for CD64, the Ka, kd, and KD of all FcRs were calculated under steady-state conditions, while the affinity for CD64 was evaluated on a 1:1 basis.

[0320] In vivo efficacy study

[0321] BALB / c SCID mice were inoculated with 10 x 10⁶ HCC1534 cells. Mice were treated with hIgG1 (10 mg / kg), CLN-619 (or 0.03, 0.3, 3.0, or 10 mg / kg), or CLN-619 DANA (0.03, 0.3, or 3.0 mg / kg). Administration began on the same day as tumor cell implantation, twice weekly for 8 doses. BALB / c mice were also subcutaneously inoculated with 5 x 10⁶ PLC / PRF / 5 cells. Mice were administered the intraperitoneal (IP) medium (PBS) or CLN-619 (0.3 mg / kg or 3 mg / kg) three times weekly for 16 doses.

[0322] For both studies, tumor size and body weight were measured twice weekly during the study period. The HCC1534 and PLCR / PRF / 5 studies ended on day 35 and day 39, respectively. Statistical analysis was performed using univariate ANOVA and multiple comparisons (GraphPad).

[0323] Treatment with luciferase-labeled human colorectal cancer cells expressing HCT-116 MICA / B was initiated on day 4 by intraperitoneal injection of hIgG1 or CLN-619 at a dose of 10 mg / kg twice weekly for five and six weeks, respectively. Bioluminescence imaging was performed twice weekly to monitor the progression of disseminated disease. Survival analysis was performed on day 40, when the last mouse reached a near-death state. Statistical analysis was performed using the log-rank test.

[0324] Pharmacokinetics

[0325] BALB / c SCID mice were administered CLN-619 by intraperitoneal injection at 1, 3, or 10 mg / kg, and blood samples were collected via mandibular sampling at the following time points during the study period: 1, 3, 6, 24, 48, 72, 96 hours and 11, 14, 21, 28 days. A total of 3 or 4 mice were evaluated at each time point in each dose group. Serum concentrations were measured by MICA / B sandwich ELISA.

[0326] B. Result

[0327] CLN-619 binds to the α3 domain of MICA / B with high affinity but does not impede interaction with NKG2D on NK cells. CLN-619 increases cell surface expression levels of MICA / B while decreasing in vitro levels of soluble MICA / B. Treatment of cancer cell lines with CLN-619 induced antibody-dependent cytotoxicity (ADCC) and antibody-dependent phagocytosis (ADCP), with EC50 values ​​dependent on MICA / B expression levels. CLN-619 potently inhibited tumor growth in mouse liver and lung xenograft models and increased survival in mice with disseminated colorectal cancer.

[0328] CLN-619 combines with the α3 domain of MICA / B

[0329] CLN-619 is derived from a parental mouse antibody selected for its high affinity binding to the α3 domain of MICA and its ability to enhance MICA / B expression on the cell surface. The variable domain of the parental antibody was humanized and introduced into the human IgG1 backbone to generate CLN-619.

[0330] The MICA / B genes are highly polymorphic, with nearly 150 MICA alleles and 50 MICB alleles existing in the human population. Typical MICA *001 and MICB *004 alleles share 83% identity in the overall protein sequence and 91% identity in the α3 domain. Given this high protein polymorphism, confirming the broad reactivity of CLN-619 is crucial. The monovalent affinity of CLN-619 for the extracellular domain (ECD) of several common human MICA alleles (MICA *001, *002, *004, and *008) and the MICB allele *004 was measured using Octet. The KD values ​​for CLN-619 binding to recombinant MICA allele variants ranged from 0.77 to 2.04 nM, all within three-fold (i.e., within 2.3 times). Figure 1AThe KD value for CLN-619 binding to the MICB allele *004 was 11.37 nM, which is approximately 5 to 15 times lower than the KD value for binding to the MICA allele. ELISA experiments independently confirmed the binding of CLN-619 to representative allele variants and demonstrated similar binding affinities between the 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 prevalent alleles. Figure 1C CLN-619 showed similar affinity for all alleles, except for the MICA allele *046, a rare allele present in less than 0.01% of the population, in which reduced binding was observed relative to other alleles. Figure 8 Overall, CLN-619 showed broad reactivity across all tested MICA alleles, which comprised 89% of the population.

[0331] X-ray crystallography was performed to elucidate the interaction between CLN-619 and the α3 domain of MICA at atomic resolution. The co-crystal structures of the CLN-619 Fab fragment and the MICA allele *001's MICA α3 domain were obtained at 2.12 Å resolution. CLN-619 recognizes discontinuous epitopes on the α3 domain, defined as Thr204 to Ser297, interacting with a total of 19 amino acid residues covering approximately 20% of the α3 domain. Notably, several known cleavage sites within the MICA / B α3 domain overlap with the epitopes recognized by CLN-619. The CLN-619 epitopes are highly conserved in the most common MICA allele variants, with only 4 amino acid variations observed among the 19 interacting amino acid residues, most of which are relatively conserved. Figure 10 Importantly, the region and angle of the α3 domain of CLN-619 binding to MICA do not spatially interfere with the binding of the α1 and α2 domains to the NKG2D dimer. Figure 2 Supporting crystallographic analysis, CLN-619 does not hinder NKG2D-MICA interactions in flow-based measurements. Figure 11 Interestingly, when MICA was combined with CLN-619, increased NK cell binding was observed compared to MICA alone, which may reflect the simultaneous involvement of CLN-619 with NKG2D and CD16A on NK cells. When anti-NKG2D antibody was included in the experimental conditions, the binding of MICA to primary NK cells was significantly reduced.

[0332] CLN-619 enhances the cell surface expression of MICA / B on tumor cells.

[0333] Based on structural data, the inventors hypothesized that the binding of CLN-619 to MICA / B in the α3 domain might prevent proteases from approaching the MICA / B ligand, thereby leading to reduced shedding and increased cell surface expression. We evaluated the ability of CLN-619 to regulate cell surface and soluble MICA / B levels in cell-based assays using human cancer cell lines HCC1534 (lung cancer), PLC / PRF / 5 (liver cancer), and HCT-116 (colorectal cancer). These cancer cell lines expressed approximately 58,000, 17,000, and 10,000 copies of MICA / B, respectively, on their cell surfaces, as measured by flow cytometry. Cell surface levels of MICA / B were quantified by flow cytometry after treatment with CLN-619, and the levels of sMICA / B present in the culture medium were measured by ELISA. CLN-619, rather than control hIgG1, enhanced the surface expression of MICA / B. Figure 3A ), while reducing sMICA levels in a dose-dependent manner. Figure 3B For the HCC1534, PLC / PRF / 5, and HCT-116 cell lines, the EC50 values ​​for increased MICA / B surface expression were 54, 69, and 51 ng / mL CLN-619, respectively. For these three cell lines, the IC50 values ​​for inhibiting sMICA / B release into the cell culture medium were 107, 162, and 79 ng / mL CLN-619, respectively. Notably, the EC50 value of CLN-619 enhancing cell surface MICA / B was similar to the IC50 value of reducing sMICA / B in the supernatant, supporting a causal relationship.

[0334] CLN-619 mediates NK cell-mediated cytokine production and target cell lysis.

[0335] The ability of CLN-619 to stabilize MICA / B ligands on the surface of tumor cells suggests that CLN-619 may enhance NK cell activation by increasing the level of ligands involved in activated NKG2D receptors. 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 The EC50 value for target cell lysis was 1-2 ng / mL. Figure 4B The activity depends on the complete Fc domain.

[0336] A silenced Fc form of CLN-619, termed CLN-619 DANA, was developed to evaluate the contribution of FcγRγ to CLN-619-mediated NK cell activation and tumor cell killing. CLN-619 DANA contains two mutations, D265A and N297A, in the Fc domain, which have previously been shown to eliminate FcγR binding to IgG1 antibodies. Although the DANA mutation causes CLN-619 to eliminate FcγR binding, it retains its ability to bind to MICA and regulate cell surface levels. Figure 9 , Figure 12 When evaluated in primary NK cell co-culture assays, CLN-619 DANA did not induce significant IFN-γ levels. Release or target cell lysis ( Figure 4A , 4B These results demonstrate that CLN-619-mediated immune activation and NK cell-induced target cell lysis require the intact Fcγ1 domain.

[0337] CLN-619 induces ADCC and ADCP in tumor cells expressing MICA / B.

[0338] Given that CLN-619 is a human IgG1 antibody with a functional Fc domain, we evaluated whether CLN-619 could mediate various Fc-mediated functional activities, such as ADCC, ADCP, and CDC. The functional involvement of FcγRIIIa / CD16A mediated by CLN-619 was investigated in a cell-based reporter assay designed as an alternative to assessing ADCC activity. The Jurkat cell line, whose luciferase gene expression is driven under the control of NFAT transcription factors in response to human FcγRIIIa / CD16A activation, was used as the reporter effector cells. Jurkat cell lines (one expressing a high-affinity V158 variant of CD16A and the other a low-affinity F158 variant) were co-cultured with target cells expressing MICA / B in the presence of CLN-619. The strongest CD16A signaling was observed after co-culturing with HCC1534 (a tumor cell line expressing high levels of MICA / B). Figure 5 With CLN-619 treatment, EC50 values ​​ranged from 8 to 81 ng / ml, with the F variant consistently showing a lower signal compared to the V variant of CD16A. Consistent with its functionally deficient Fc domain, CLN-619 DANA was inactive in Jurkat cell assays. Overall, the EC50 value of CD16A activated by CLN-619 in reporter cell lines correlated with MICA / B expression levels on target cells, suggesting that CLN-619 can mediate ADCC in a target-dependent manner.

[0339] Given that CLN-619 can also bind to FcγRIIA / CD32A (the FcγR receptor primarily responsible for ADCP), the ability of CLN-619 to induce macrophage-mediated phagocytosis was also evaluated

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

[0340] Although CLN-619 can mediate ADCP and ADCC, no complement-dependent cytotoxic (CDC) activity was observed in response to CLN-619 treatment (data not shown). However, in co-culture assays with primary macrophages, CLN-619 treatment induced dose-dependent phagocytosis of labeled MICA / B-expressing tumor cells.

[0341] Antitumor activity of CLN-619

[0342] 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 using MICA / B-expressing human tumor xenografts in immunodeficient BALB / cSCID mice. In BALB / c SCID mice, exposure following intraperitoneal (ip) administration of CLN-619 was dose-proportional, and the serum half-life was consistent with that of typical human IgG1 antibodies administered in mice.

[0343] In the PLC / PRF / 5 liver tumor xenograft model, compared with the control group, administration of CLN-619 at 0.3 mg / kg or 3 mg / kg g.p. induced complete inhibition of tumor growth (p=0.015). Figure 7A In the HCC1534 lung xenograft model, CLN-619 administered intraperitoneally 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 homologous control mAbs, with tumor growth inhibition rates (TGI) ranging from 45% to 87%. No antitumor activity was observed in CLN-619 DANA cases at any dose level. Figure 7BThis data indicates that the intact Fcγ1 domain of CLN-619 is essential for its in vivo antitumor activity, consistent with our in vitro observations in NK cell co-culture assays.

[0344] The activity of CLN-619 in a metastatic setting was explored in a mouse model designed to mimic advanced disseminated disease. HCT-116 human colorectal cancer cells expressing luciferase were inoculated into mice via intraperitoneal injection (ip) to induce tumor cell dissemination to distant organs. Kaplan-Meier survival analysis demonstrated that CLN-619 treatment resulted in a statistically significant survival benefit compared to hIgG1 (p<0.0001). Figure 7C ).

[0345] C. Conclusion

[0346] CLN-619 inhibits the shedding of MICA / B, effectively restoring cytotoxic signaling pathways in immune cells. Potent antitumor activity of CLN-619 as a monotherapy has been observed in several preclinical models. The activity of CLN-619 requires a functional Fcγ1 domain, suggesting that the simultaneous involvement of NKG2D and CD16A on immune cells is necessary to achieve optimal cytotoxicity. The preclinical data reported in this example support the evaluation of CLN-619 in cancer patients.

[0347] The data presented in this paper reveal the multimodal mechanism of action of CLN-619. In addition to inhibiting MICA / B shedding and increasing NKG2D ligand expression on cancer cells, we also observed that the ADCC and ADCP functions of CLN-619 depend on the wild-type hIgG1 backbone. Despite retaining NKG2D binding ( Figure 11 However, the Fc-deficient form of CLN-619 did not show activity in the NK co-culture killing assay. Figure 4B CLN-619 DANA also lacks activity in vivo. We hypothesize that dual stimulation of the CD16A and NKG2D receptors on NK cells may be necessary for CLN-619 activity. This concept of dual receptor involvement is supported by published studies showing that NKG2D involvement can synergistically enhance ADCC and strengthen CD16A and NKp46 activation in resting NK cells.

[0348] CLN-619 demonstrated remarkable single-agent activity at low dose levels in various tumor xenograft models, using tumor cell lines representing indications where the NKG2D pathway has proven clinically relevant, such as HCC and NSCLC. The human xenograft model was chosen because mice lack orthologs of MICA / B, and mouse NKG2D on NK cells has been shown to recognize MICA / B on human tumor cells. The host mice used in our study possessed active NK cells and macrophages but lacked T cells. Therefore, the observed antitumor efficacy is likely attributable to NK cell lysis of target cells and / or phagocytosis from macrophages. The contribution of NKG2D-expressing T cell subtypes (such as CD8+ T cells, NKT cells, and γδ T cells) to efficacy was only observed in the fully immunized model. Notably, the NKG2D axis has been shown to play a co-stimulatory role in lowering the CD8+ T cell lysis threshold, even driving CD8+ T cell-mediated cancer cell lysis after prior TCR activation in the absence of MHC expression. CLN-619 may engage multiple cytotoxic immune effector cells in the tumor microenvironment (TME) to enhance antitumor activity. In a co-culture system containing an enriched population of CD8+ T cells and tumor cells expressing MICA / B, CLN-619 induced dose-dependent release of IFNγ, upregulation of the CD25 marker for T cell activation, and target cell lysis. Figures 32A-32C ).

[0349] Therapies that potently engage NK cells may be more advantageous than those that engage only T cells. NK cells do not require antigen-induced activation, do not secrete high levels of cytokines that could trigger CRS, and do not induce graft-versus-host disease (GVH). However, NK cells may require more than one activating stimulus to fully unleash their cytotoxic potential and overcome inhibition from the many negative regulatory receptors expressed on NK cells. Furthermore, NK cells are unevenly distributed in the tumor microenvironment (TME), with varying numbers of infiltrating NK cells across different indications. In multiple indications, dysfunctional NK cell subsets have been found to be enriched in the TME and associated with poor prognosis. One mechanism for improving NK cell performance may be the simultaneous involvement of NK cell-activated receptors, as described in this paper regarding CLN-619. In addition, combinations with therapies that enhance NK cell function, persistence, and / or tumor infiltration may be beneficial and should be further explored.

[0350] In summary, the data reported in this article support clinical studies of CLN-619 for the treatment of multiple tumor types. The following sections will describe in further detail a Phase I clinical trial (NCT05117476) of CLN-619 as a monotherapy and in combination with pembrolizumab in cancer patients.

[0351] Example 2. A phase 1 dose-escalation study aimed at investigating the effects of CLN-619 (anti-MICA / B antibody) on patients with advanced solid tumors. Safety, efficacy, pharmacokinetics, and pharmacodynamic activity in cancer patients

[0352] Background: This embodiment describes an ongoing Phase 1, multicenter, open-label, first-in-human dose-escalation study (NCT05117476) evaluating CLN-619 as monotherapy for patients with advanced solid tumors who have progressed after existing therapy, are ineligible for treatment, or refuse non-curative standard therapy. CLN-619 is a humanized MICA / B-specific IgG1 monoclonal antibody that binds to NKG2D ligands MICA and MICB and prevents the proteolytic cleavage of these ligands from tumor cells, thereby increasing the lysis of tumor cells by innate and adaptive immune cells. Table 1 provides the amino acid sequence of CLN-619. CLN-619 is expected to have broad-spectrum antitumor activity.

[0353] Methods: This study was designed to characterize the safety, tolerability, dose-limiting toxicities, and preliminary antitumor activity of intravenously administered CLN-619 monotherapy in patients with advanced solid tumors. Patients aged 18 years or older with metastatic or locally advanced solid tumors, disease progression following prior therapy with no further standard of care available, an ECOG score of 0 or 1, and an estimated life expectancy of ≥12 weeks were recruited. Eligibility criteria included measurable disease according to RECIST v1.1 and adequate liver and kidney function and hematological parameters.

[0354] The following patients were excluded from the study: (i) receiving study therapy within 25 days (or 5 half-lives) of the first planned dose of CLN-619; (ii) exhibiting severe and / or uncontrolled medical conditions, including active autoimmune diseases requiring immunosuppressive drugs; (iii) receiving treatment for an acute infection within 7 days of the first planned dose of CLN-619; (iv) having an immunological adverse event of grade ≥3 with prior checkpoint inhibitor therapy; or (v) having active CNS metastases.

[0355] CLN-619 alone is administered intravenously over one hour every three weeks (Q3W) at dose levels (DL) of 0.1 mg / kg, 0.3 mg / kg, 1 mg / kg, 3 mg / kg, 6 mg / kg, or 10 mg / kg.

[0356] Administer a standard prophylactic medication (such as acetaminophen or ibuprofen) 30–60 minutes before each CLN-619 dose and before each subsequent dose, as is customary at the institution.

[0357] A corticosteroid prodrug starting at 3 mg / kg DL is needed to prevent infusion-related reactions (IRR).

[0358] We are exploring parallel dose escalation of CLN-619 (DL 1.0 mg / kg and higher) in combination with pembrolizumab (as outlined in Example 2 below).

[0359] Response is assessed every 9 weeks (RECIST 1.1). Patients are permitted to continue treatment until disease progression, intolerable toxicity, or a maximum of 34 cycles. Patients whose disease progresses may continue treatment if clinical benefit and acceptable tolerability are determined.

[0360] Study endpoints included: adverse events (CTCAE v5.0), dose-limiting toxicities (DLT), response (RECIST v1.1), and pharmacokinetic parameters (e.g., C). max AUC 0-504h t 1 / 2 ).

[0361] This article describes data from dose escalation of a single therapy.

[0362] Results: Baseline patient characteristics were... Figure 14 As shown in the figure, and the number of patients within each dose level is Figure 15 The results are shown in the table. In short, a total of 37 patients were recruited (median age 63 years, range 26–83 years), of whom 62% were female and 60% had ECOG PS 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), adenoid cystic salivary gland 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 therapy (median 3, range 1–7); 20 patients had previously received checkpoint inhibitors. The median number of CLN-619 cycles received was 2 (range: 0–13).

[0363] Treatment-interventional adverse events (TEAEs) were monitored, and CLN-619 was observed to be well-tolerated, with most TEAEs being grade 1 / 2. TEAEs were observed in ≥10% of patients. Figure 16The adverse events (described in the text) included abdominal pain, nausea, fever, infusion-related reactions (IRR), decreased appetite, fatigue, vomiting, and back pain. IRR occurred at dose levels of 0.3 mg / kg or higher. All IRRs occurred during the first cycle and subsided within 3 hours of onset. One case of Gr3 laryngeal edema occurred at a 10 mg / kg DL in the absence of the required steroid prophylaxis, leading to discontinuation of administration. No fatal TEAEs or adverse events meeting the protocol-defined DLT criteria were reported. These data demonstrate the safety of CLN-619 in a clinical setting. Figure 17A and 17B As shown, observations were observed between the 0.1 mg / kg and 10 mg / kg dose levels, as indicated by C. max CLN-619 exposure, as measured, increased almost proportionally with the dose. Exposure was observed between 0.1 and 1 mg / kg as indicated by AUC. 0-504h The measured CLN-619 exposure was greater than the dose-proportional increase, and a nearly dose-proportional increase in CLN-619 exposure was observed between the 1 and 10 mg / kg dose levels.

[0364] For DL ​​doses ranging from 0.1 mg / kg to 10 mg / kg, the half-life of CLN-619 ranges from 61.3 to 443 hours.

[0365] Cytokine expression in longitudinal serum samples from patients treated with CLN-619 monotherapy was measured using Myriad RBM Inflammation and a custom MAP assay on the Luminex platform. Data are shown in... Figure 18 In the figure, the mean absolute cytokine level (pg / ml) at each dose level over time is depicted. Error bars illustrating the SD are shown. Figure 18 As shown, a transient increase in cytokine levels was observed 2–6 hours after the first dose of CLN-619, and returned to baseline within 4–8 days. No dose-dependent increase in cytokine secretion was observed in the periphery.

[0366] Treatment time and clinical activities Figure 19A and 19B The objective response summary and the best single-therapy response are shown in the figure. Figure 20A and 20B Overview. Confirmed complete response (CRR) has been achieved in patients with recurrent salivary gland tumors whose disease progressed after a complete response to PD-1 blockade therapy. Figure 21 ). Figure 22The data presented confirm an objective response in patients with endometrial cancer and a large tumor burden whose disease progressed after platinum-based chemotherapy and anastrozole treatment. Stable disease duration for more than three cycles was observed in seven other patients. For example, one patient with cervical cancer maintained SD for nine cycles before disease progression.

[0367] Conclusion: CLN-619 therapy was well tolerated across a dose range of 0.1 to 10 mg / kg, and no dose-limiting toxicities were observed at any administered dose, demonstrating the clinical activity of CLN-619 as a monotherapy. Furthermore, objective responses were observed in patients whose disease progressed after PD-1 therapy. No grade ≥3 TRAEs were observed in patients receiving regimen-mandated prophylactic drugs. Single-agent activity, including objective responses, was observed in multiple tumor types in both patients receiving and not receiving checkpoint therapy. Significant single-agent clinical activity was observed in multiple gynecological malignancies, demonstrating surprising efficacy in these subjects, thus prompting an expansion of the treatment consortium for endometrial and cervical cancer.

[0368] Example 3. A phase 1 dose-escalation study was conducted to investigate the effects of CLN-619 (anti-MICA / MICB antibody) alone and in combination with p-8. Safety, efficacy, pharmacokinetics, and pharmacodynamic activity of emumab combination therapy in patients with advanced solid tumors

[0369] The above embodiments demonstrate the surprising and unexpected safety and efficacy of anti-MICA / B antibodies as a monotherapy pan-cancer therapeutic agent. This embodiment further demonstrates the potential of this therapeutic agent in combination therapy. Currently, there are no approved products targeting the NKG2D ligand / receptor pathway. CLN-619 targets this key pathway and represents a novel approach in immuno-oncology, potentially benefiting cancer patients in the future.

[0370] Study Description: A phase 1, open-label, first-in-human, multicenter, dose-escalation and dose-expansion study of CLN-619 in patients with selected advanced solid tumors, administered alone (Modules A and D), in combination with pembrolizumab (Module B), or in combination with chemotherapy (Module C). For the overall study design, see [link to study description]. Figure 23A and 23B .

[0371] CLN-619 will also be administered weekly during the first cycle to study the loading dose plan.

[0372] Pembrolizumab is an anti-PD-1 antibody that will be administered to patients via IV infusion at a dose of 200 mg every 30 minutes.

[0373] The chemotherapy to be administered includes:

[0374] Carboplatin AUC 5 and pemetrexed 500 mg / m2 Q3W IV,

[0375] Carboplatin AUC 5-6 Q3W and paclitaxel (175 mg / m2 Q3W) IV, and

[0376] Paclitaxel (60-80 mg / m2 Q1W) IV.

[0377] Clinical research:

[0378] As of June 22, 2023, CLN-619 was the only antibody targeting MICA / MICB to be in clinical trials. In the current study, a first-in-human study of CLN-619, 61 patients were administered multiple doses of CLN-619, alone or in combination with pembrolizumab, every three weeks (maximum dose 10 mg / kg). CLN-619 was well-tolerated in both monotherapy and combination therapy scenarios, with no dose-limiting toxicities observed during dose escalation and no grade 3 treatment-related adverse events (Ae) in patients receiving protocol-mandated progenitor drugs. At the time of analysis, a total of 235 doses of CLN-619 had been administered, including 52 doses in combination with pembrolizumab. Expanded-group studies as monotherapy in cervical and endometrial cancers have begun and are currently underway.

[0379] Pembrolizumab. Pembrolizumab is marketed in the United States as Keytruda. ® Pembrolizumab is a humanized IgG4 anti-PD-1 monoclonal antibody manufactured by Merck for the treatment of cancer. Pembrolizumab acts as an immunomodulator by blocking the interaction between the programmed cell death 1 (PD-1) receptor on activated T cells and its ligands PD-L1 and PD-L2 (expressed on tumor cells and immune cells).

[0380] Basis for the combination of CLN-619 and pembrolizumab

[0381] There is strong evidence to support the coordinated targeting of innate and adaptive immunity by combining CLN-619 and pembrolizumab. CLN-619 enhances NKG2D-mediated killing by NK and selected T cell populations and also enhances antibody-dependent cytotoxicity, a key cytolytic mechanism mediated by NK cells and other innate immune cell populations. Pembrolizumab enhances T cell-mediated immunity by disrupting T cell exhaustion mediated by PD-1 and its ligands PD-L1 and PD-L2. Recently, there has been evidence that the PD-1 / PD-L1 axis may also play a role in regulating NK cell function (Dunai and Murphy 2018). Therefore, both CLN-619 and pembrolizumab can enhance both innate and adaptive immunity through their respective mechanisms of action.

[0382] In the clinical setting, data suggest the potential for synergistic effects between CLN-619 and checkpoint inhibitors (CPIs). For example, in melanoma patients treated with CPIs, patients with detectable levels of sMICA / sMICB had lower overall survival (OS) compared to those who were sMICA / sMICB negative (Maccalli 2017). CPI therapy may be more effective if MICA / MICB levels on the tumor surface are stabilized through CLN-619 treatment. Furthermore, it has been hypothesized that sustained NKG2D signaling can lead to a PD-L1-rich tumor microenvironment, suggesting that NKG2D modulatory therapies would benefit from combination with PD-1 pathway inhibitors (Sheppard 2018).

[0383] Chemotherapy

[0384] There are various types of chemotherapy, each with a different mechanism of action. The types of chemotherapy to be administered during this study are briefly described below.

[0385] Platinum preparation (carboplatin) Platinum compounds exert their cytotoxic effects through two main mechanisms. They are alkylating agents that bind to and cross-link DNA strands, thereby inhibiting DNA synthesis and function. If the DNA is sufficiently damaged, the cell undergoes apoptosis. Platinum chemotherapeutic agents can also induce an immunogenic, DNA-independent cell death process. They interact with many cellular proteins, thereby regulating several signal transduction pathways.

[0386] Taxanes (paclitaxel) Taxanes exert their anticancer effects by binding to and preventing the depolymerization of the microtubule spindle system. This leads to blockage of the metaphase-anaphase transition and ultimately inhibits mitosis and induces apoptosis. Unlike other microtubule-disrupting drugs (such as vinca alkaloids), paclitaxel stabilizes microtubules by binding to polymerized tubulin, thereby preventing tubulin degradation. The broad-spectrum activity of paclitaxel is predicted by this mechanism of action (i.e., control of cell proliferation and DNA repair), which targets essential elements of the cancer phenotype.

[0387] Pemetrexed Pemetrexed is a novel, multi-targeted antifolate drug that inhibits enzymes involved in folate metabolism and the synthesis of purines and pyrimidines. These precursors prevent the formation of DNA and RNA essential for the growth and survival of both normal and cancer cells.

[0388] The basis for combining CLN-619 with chemotherapy

[0389] There is ample evidence to support the combination of CLN-619 with standard-of-care chemotherapy. Cytotoxic chemotherapy agents such as platinum-based, taxane-based, and folic acid antimetabolites have been shown to induce cellular stress, leading to the upregulation of NKG2D ligands, including MICA / B. Furthermore, these agents have been shown to have positive effects on the immune response, including increasing immune cell infiltration and effector cell activity while decreasing the level and function of suppressor immune cells. Due to their respective mechanisms of action, synergistic effects may exist between CLN-619 and chemotherapy agents.

[0390] Platinum-based agents such as carboplatin and cisplatin have been shown to enhance MICA / B expression, as illustrated in a small study of carboplatin-treated ovarian cancer patients and preclinical studies of cisplatin-treated NSCLC cell lines (Capellero 2020; Okita 2019). These agents have also been shown to actively modulate immune responses in preclinical models and some clinical settings. In preclinical models, cisplatin treatment led to the recruitment / proliferation of effector cells; upregulation of the lysogenic activity of cytotoxic effectors; and reduction of Tregs in the TME (Gameiro 2012; de Biasi 2014). Although carboplatin treatment had no effect on NK cells, in in vitro studies, carboplatin treatment of ovarian cancer patients increased the ability of CD8+ T cells to produce IFNγ (Wu 2009).

[0391] 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 agents have also been shown to have positive effects on immune function in preclinical models and patients. In preclinical models and breast cancer patient samples, docetaxel treatment has been shown to increase NKG2D on NK cells, and in the case of preclinical models, docetaxel-mediated increased responsiveness to trastuzumab via NKG2D (Di Modica 2016). Increased CD8+ T cell infiltration into tumors has been observed in NSCLC patients treated with docetaxel (Gao 2019). Finally, paclitaxel has been shown to reprogram tumor-associated macrophages to a more M1 or immunogenic phenotype in preclinical and patient samples (Wanderley 2018).

[0392] While the folic acid antimetabolite 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, 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 combined with PD-1 blockade, it caused significantly enhanced antitumor activity in preclinical models (Schaer 2019).

[0393] In addition to the data above highlighting the role of chemotherapy in actively modulating the MICA-NKG2D pathway and immune effector activity, chemotherapy combined with immunotherapy has been extensively tested in various tumor types in conjunction with anti-PD(L)1 therapy. Pembrolizumab in combination with chemotherapy (including platinum-based and taxane-based) has been approved for the treatment of multiple tumor types, such as NSCLC, endometrial cancer, breast cancer, and gastric cancer (pembrolizumpi 2021). Dotalimab has also been approved in combination with platinum-based chemotherapy for first-line treatment of patients with mismatch repair deficient (dMMR) advanced endometrial cancer (N Engl J Med 2023;388:2145-58. DOI: 10.1056 / NEJMoa2216334). While the combination of anti-PD(L)1 and chemotherapy has been successful in some solid tumors, they have not yet been shown to be effective in other solid tumors, such as epithelial ovarian cancer or target-driven NSCLC, such as those solid tumors with EGFRm or ALK rearrangements.

[0394] Cytotoxic chemotherapy can produce immunomodulatory effects, such as disrupting 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 of using this combination in several cancer types have been reported, including improved survival. Given the mechanisms by which cytotoxic agents alone induce cellular stress and MICA / B expression, there is strong clinical evidence suggesting that combining chemotherapy with CLN 619 may have synergistic effects, which will be explored in this study.

[0395] Basis for starting dose and dosing schedule

[0396] Allometric scaling was used to calculate the FIH dose and dosing regimen of CLN-619 to estimate human PK and expected receptor occupancy (RO), as well as in vivo and in vitro safety and pharmacological data. Using this method, a starting dose of 0.1 mg / kg and a Q3-week dosing schedule are recommended for CLN-619 clinical trials. Based on all available toxicological and pharmacological data, the expected starting dose is safe and remains within the lower range of CLN-619's pharmacological activity.

[0397] Human PK parameters were determined based on allometric scaling from cynomolgus monkey PK data. The calculated expected clearance rate in humans is 0.19 L / day. The expected terminal half-life in humans is 26.9 days, which is typical for a monoclonal antibody, demonstrating the rationality of the chosen Q3W dosing schedule.

[0398] The NOAEL (101.4 mg / kg / week) in a one-month repeated-dose monkey toxicity study supports 0.1 mg / kg as a safe starting dose. Following a single dose of CLN-619 at the recommended starting dose of 0.1 mg / kg, the predicted area under the curve (AUC0-168) and Cmax in cynomolgus monkeys were 775-fold and 1,020-fold lower, respectively, than at the highest non-serious toxicity dose (equivalent to the NOAEL), thus providing a large safety margin.

[0399] No findings were observed after administration of CLN-619 to monkeys, consistent with the target biology. Notably, MICA / MICB expression was highly restricted in normal tissues. MICA / MICB expression is induced under stress conditions (e.g., infection, radiation, malignancy), a normal physiological process that triggers immune cells to eliminate cells expressing MICA / MICB. In healthy monkeys, normal tissue expression levels of MICA and MICB are not high; therefore, the effects of CLN-619 treatment on normal animals are expected to be minimal.

[0400] Using modeling methods, the expected RO of CLN-619 for MICA and MICB was determined using the in vitro association rate constant and equilibrium dissociation constant. The total target occupancy of MICA / MICB is expected to be approximately 30% at the starting dose of CLN 619. An expected RO of 30% is considered reasonable based on the anticipated administration of CLN 619 to patients with advanced cancer, where the aim is to start at the pharmacologically active dose (PAD), but patient safety is also taken into consideration.

[0401] In vivo and in vitro pharmacological data were also considered in determining the starting dose of CLN-619. Based on the in vivo and in vitro pharmacological data, the recommended starting dose of CLN-619 was expected to be in the lower range of pharmacological activity. At the recommended starting dose of 0.1 mg / kg, administered by 1-hour infusion, the expected human Cavg was approximately 5-fold lower than the Cavg measured in mice at the effective dose in in vivo xenograft mouse studies. For in vitro functional studies measuring the target regulation and Fc effector function of CLN-619 in tumor cell lines, EC50 values ​​ranged from 0.045 to 0.1 µg / mL. However, given that the expected tumor exposure level is 10-fold lower than the circulating level, leading to an overestimation of drug efficacy in tumors, the correlation between in vitro data and FIH dose determination needs to be treated with caution. Furthermore, while in vitro functional assays can technically demonstrate mechanism-driven activity, the sponsor believes that these assessments have limited value because in vitro assays are closed, static systems and are not expected to represent the open, dynamic in vivo condition, in which CLN-619 is gradually cleared over time.

[0402] In vitro cytokine release assays further support the expectation that the starting dose is pharmacologically active and safe. At all tested concentrations, treatment with CLN-619 using human PBMCs (6 donors) failed to elicit a cytokine / chemokine response above background levels in either soluble or wet-bound form. In the case of co-culture systems of PBMCs with MICA / MICB-expressing cell lines, the CLN-619-induced release-limited group was associated with cytokines considered to have pharmacodynamic effects on the target, including MIP1α, TNFα, and IL-8. In the co-culture system, TNFα levels in the 6 donors were 16 to 210 times lower than in the positive control at concentrations 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 pivotal one-month in vivo monkey toxicity study, the risk of cytokine release at the clinical starting dose of CLN 619 is low.

[0403] Based on these data, the initial dose of 0.1 mg / kg and the Q3W dosing schedule were chosen as safe and still within the lower range of pharmacological activity of CLN-619.

[0404] Preliminary analysis of CLN-619 PK data collected at dose levels ranging from 0.1 mg / kg to 10 mg / kg indicates that, at least for doses from 1 mg / kg to 10 mg / kg, the increase in exposure is dose-proportional. Therefore, at these dose levels and above, the PK of CLN-619 appears to be linear, and thus an increase in total exposure would be analogous to an increase in total dose. At higher dose levels, the mean half-life is as high as 19 days, suggesting that steady-state exposure may be reached in 60 to 90 days, or approximately 3 to 6 CLN-619 cycles.

[0405] To achieve steady-state exposure more quickly while limiting the maximum serum concentration of CLN-619, a loading dose strategy can be employed in the first cycle. The planned loading dose regimen would administer CLN-619 at a dose of 10 mg / kg three (3) times per week during the first cycle. On day 1 of the second cycle, an additional 10 mg / kg dose would be administered, and from the second cycle onwards, CLN-619 would be administered according to the Q3W regimen. As stated above, the total increase in dose will be proportional to the total increase in exposure. Therefore, the total exposure (AUC) will increase approximately 3-fold in the first cycle compared to the first cycle under the 10 mg / kg Q3W regimen, which has demonstrated good tolerability. However, the total exposure is not expected to significantly exceed the steady-state exposure observed under the 10 mg / kg Q3W regimen during this period. With the loading dose, steady-state exposure is expected to be reached by the end of the first cycle, which may provide a greater likelihood of achieving a rapid response.

[0406] Given that the loading dose is not expected to reach a level higher than the previously observed systemic exposure at the current steady state of 10 mg / kg Q3W, this dose increase is expected to be safe.

[0407] Safety and risk / benefit assessment

[0408] As of June 22, 2023, 61 patients had received CLN-619 at doses of 0.1, 0.3, 1, 3, 6, and 10 mg / kg. Forty-three patients had received CLN-619 as monotherapy, with 12 receiving a 10 mg / kg dose. Eighteen patients had received CLN-619 in combination with pembrolizumab at doses of 1, 3, 6, and 10 mg / kg, with 6 receiving a 10 mg / kg dose. In total, 23 cases of treatment-associated sepsis (SAE) were reported following CLN-619 administration (monotherapy and in combination with pembrolizumab). One serious adverse event related to CLN-619, namely infusion-related reaction (IRR) and laryngeal edema, was reported in a patient in the absence of a mandatory steroid prophylaxis. One non-serious grade 3 rash was reported in monotherapy (Module A). No serious adverse events to CLN-619 were reported in the combination therapy (Module B), but one case of acute kidney injury (AKI) SAE was considered likely to be related to pembrolizumab (not to CLN-619). No adverse events (Aes) met the regimen-defined dose-limiting toxicity (DLT) criteria, and no ≥ Grade 4 treatment-associated adverse events (TEAEs) occurred. To date, the most common treatment-associated adverse events (TEAEs) in ≥ 5% of patients treated with CLN-619 as monotherapy included IRR (23.3%), abdominal pain (18.6%), fatigue (18.6%), and fever (18.6%). To date, the most common treatment-associated adverse events in ≥ 5% of patients treated with CLN-619 in combination with pembrolizumab included 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 Investigator's Handbook (IB) provides more detailed safety information.

[0409] The estimated frequency of IRR at the first dose was 21.3% (occurring in 13 of 61 patients treated). The estimated frequency at any dose was 6.3% (occurring in 15 of 235 CLN-619 administrations). Most IRRs (13) occurred during or immediately after the first dose; one IRR occurred after the sixth dose; and one patient had an IRR after both the first and second doses. One event occurred in patients receiving combination therapy with pembrolizumab. The number of patients receiving combination therapy remains limited (18), making it difficult to assess whether this addition alters the IRR risk. While all patients recovered without sequelae, some patients may have experienced severe IRRs.

[0410] This study initially selected a 60-minute infusion time and IV administration, supported by non-clinical toxicology studies in which CLN-619 was administered once weekly via a slow IV bolus over 5 minutes. Based on clinical experience in the dose escalation group, the infusion time for the first 1-2 infusions was extended to 120 minutes to reduce the frequency and severity of IRR.

[0411] CLN-619-001 is the first-in-human clinical trial of CLN-619, both alone and in combination with pembrolizumab (an anti-PD-1 antibody), and will be conducted in patients with locally advanced or metastatic solid tumors. Patients in Module B must have tumor types listed in their current prescribing information for pembrolizumab. Restrictions on prior treatment history are described in the inclusion criteria. Unless otherwise specified in the inclusion criteria, patients will be required to have received, refused, or demonstrated contraindications to or intolerance to their approved standard therapy, or have other approved treatments. CLN-619-001 utilizes a modular study design, where Module A is designed to investigate the safety, tolerability, and preliminary anti-tumor activity of CLN-619 as monotherapy, while the branch design will support Module B in investigating the combination of CLN-619 and pembrolizumab (an anti-PD-1 antibody) (see [link to study study]). Figure 23B Module A monotherapy dose escalation will evaluate the safety of multiple escalation dose levels of CLN-619 to determine the maximum tolerated dose (MTD) and recommended phase 2 dose (RP2D) for CLN-619 monotherapy. Assuming acceptable safety and tolerability with respect to dose escalation for CLN-619 monotherapy, Module A cohort expansion will be initiated to recruit tumor-specific cohorts of patients with non-small cell lung cancer (NSCLC), cervical cancer, and endometrial cancer. Up to three additional patient cohorts with specific tumor types may be recruited to Module A cohort expansion based on evidence of clinical benefit and / or any observed relationships and objective responses between hematologic and tumor-based biomarker data observed during the study, or other applicable emerging data. These cohorts will be further evaluated for the pharmacodynamics (PD), safety, and initial antitumor activity of CLN-619 monotherapy.

[0412] The branch trial design will also support the initiation of a dose-escalation cohort of Module B combination therapy in patients with advanced solid tumors. This cohort will use a 3+3 design to investigate the safety, tolerability, and initial antitumor activity of the combination of CLN-619 and pembrolizumab. Once the safety of a 3 mg / kg monotherapy dose is established, the dose-escalation cohort of Module B combination therapy can be initiated at a 1 mg / kg CLN-619 dose. If the CLN-619 monotherapy MTD is less than 3 mg / kg, the dose-escalation cohort of Module B combination therapy can be initiated at a dose level below the MTD. Pembrolizumab (200 mg) will be administered immediately prior to CLN-619. A standard 3+3 dose-escalation protocol will be followed. Planned dose levels are 1.0, 3.0, 6.0, and 10 mg / kg.

[0413] Following demonstration of the acceptable safety and tolerability of the CLN-619 plus pembrolizumab combination, Module B expansion will evaluate the preliminary safety and antitumor activity of the CLN-619 plus pembrolizumab combination in patients with NSCLC and endometrial cancer. Up to three additional disease-specific expansion cohorts may be recruited to Module B expansion based on evidence of clinical benefit and / or any observed relationship between blood and tumor-based biomarker data, observed objective response, stable disease, or other applicable emerging data. The monotherapy dose-escalation cohort and the combination therapy dose-escalation cohort will recruit patients in a conservative, stepwise manner under close supervision, guided by standard decision-making rules.

[0414] Module C will explore the safety, tolerability, and preliminary efficacy of combination chemotherapy with standard care in predefined tumor types. The combination dose expansion cohort will be recruited in a conservative, stepwise manner, under close monitoring, guided by standard decision-making rules.

[0415] Adding Module D, the sponsor will investigate whether the loading dose administered weekly in the first cycle reaches steady-state exposure at the end of the first cycle, which may offer a greater likelihood of achieving a rapid response. Given that the loading dose is not expected to reach systemic exposure higher than previously observed at the currently well-tolerated steady-state of 10 mg / kg Q3W, this dose increase is expected to be safe. Taken together, this study provides a reasonable risk-benefit assessment for patients with advanced cancer and limited treatment options.

[0416] Module A: Single Therapy Dosage Escalation

[0417] Patients with advanced selected solid tumors will be treated with escalating doses of CLN-619 monotherapy, initially using a single-patient accelerated titration design. Once the first patient recruited at a given dose level experiences a ≥ Grade 2 CLN-619-related (defined as probable or definitively related) adverse event (AE) at a dose level of 3.0 mg / kg (whichever occurs first) within the first three-week treatment cycle, transitioning to a standard 3+3 dose escalation regimen. The planned dose levels are shown in Table 3 below.

[0418] Table 3: Monotherapy Dosage in Module A During Incremental Period

[0419]

[0420] Data obtained during dose escalation in Module A will be used to determine the MTD (or MAD, if no MTD is defined) for CLN-619 monotherapy. Alternatively, MBED may be specified as a dose below the MTD based on a review of PK / PD, safety, and efficacy data, and may also guide dose selection for expanded Module A groups (i.e., RP2D).

[0421] Module A: Single Therapy Dosage Escalation Dosage Level Group Expansion

[0422] The purpose of expanding the recruitment of these dose level groups is to further characterize the safety, PK, PD, biomarkers and clinical activity of CLN-619, as well as to characterize the clinical and biological activity of the selected dose levels for expansion.

[0423] The plan was to recruit up to 10 patients with advanced solid tumors in each cohort expansion at the selected dose level (including patients recruited from the original dose level cohort). A total of 45 patients were recruited into the 7 cohorts.

[0424] Research procedures and event planning in group expansion ( Figure 24 ) and sampling plan ( Figure 25 The procedure is the same as that described in the original group, except that two fresh biopsy samples must be collected at baseline (screening) and after the first dosing cycle (i.e., C2D8 -1 day / +3 days).

[0425] Module A: Single Therapy Group Expansion

[0426] Following the selection of CLN-619 monotherapy for RP2D, Module A (Monotherapy Group Expansion) will be initiated. This module will investigate the safety and initial antitumor activity of CLN-619 monotherapy and recruit patients to the tumor-specific expansion group.

[0427] • Expanded A1: CLN-619 monotherapy in patients with non-small cell lung cancer (NSCLC) (N=16 patients).

[0428] • Expanded A2: CLN-619 monotherapy for cervical cancer patients (N=16 patients).

[0429] • Expanded A3: CLN-619 monotherapy at dose level #1 for patients with endometrial cancer (N=10 patients).

[0430] • Expanded A4: CLN-619 monotherapy at dose level #2 for patients with endometrial cancer (N=10 patients).

[0431] • During the dose escalation phase of the study, up to three additional disease-specific expansion cohorts may be declared based on evidence of clinical benefit (objective response or stable disease throughout at least six treatment cycles) for that tumor type. Alternatively, these additional tumor-specific expansion cohorts may be triggered by considering the relationship between hematologic and tumor-based biomarker data and objective response.

[0432] • Expand A1 and A2: If 2 out of 16 patients experience clinical benefit (defined as an objective response or stable disease throughout at least 6 treatment cycles), the cohort can be further expanded to recruit up to 40 patients in total to further characterize the preliminary antitumor activity of CLN-619 monotherapy in this cohort.

[0433] • Expanding A3 and A4: Each cohort will initially be expanded to 10 patients, including those who received the drug at the corresponding dose level during the dose escalation phase. If at least one objective response is observed in the first 10 patients treated, additional patients may be recruited, up to a maximum of 40 endometrial cancer patients (20 per dose level, including those who received the drug during the dose escalation phase).

[0434] • Additional disease-specific expanded cohorts: Each cohort will initially expand to 10 patients. Additional patients may be recruited if at least one of the first 10 patients treated has evidence of clinical benefit (objective response or stable disease throughout at least 6 treatment cycles), up to a maximum of 40 patients with a defined tumor type (including those who received treatment during dose escalation).

[0435] Module B combination therapy dose escalation

[0436] Module B combination therapy dose escalation will be initiated to investigate the safety, tolerability, and initial antitumor activity of the CLN-619 combination with pembrolizumab. These cohorts will recruit patients with tumor types described in the current pembrolizumab prescribing information. After demonstrating the safety of the 3 mg / kg monotherapy dose level, cohorts may be recruited concurrently with the Module A monotherapy expansion cohort, starting at dose level 1 (1 mg / kg). If a lower dose is determined as the MTD in the Module A monotherapy dose escalation group, dose levels below the MTD will be investigated in combination with pembrolizumab.

[0437] The planned dose levels are shown in Table 4 below.

[0438] Table 4: Dosage of Module B Combination Therapy During Incremental Period

[0439]

[0440] 1 Administer pembrolizumab immediately before CLN-619

[0441] 2 Once the SRC has determined safety in Module A, Module B combination therapy dose level 1 (1 mg / kg) will be initiated with a CLN-619 dose of 3.0 mg / kg. Dose levels -1 and -2 can be evaluated as described in the decision rule.

[0442] If no DLT is observed in the first three patients at dose level 1 within the first three-week cycle, CLN-619 will be increased. If a DLT occurs as noted in the decision rule, the dose can be decreased at the dose levels listed in Table 4.

[0443] Module B combination therapy group expansion

[0444] After demonstrating acceptable safety and tolerability of the CLN-619 combination with pembrolizumab, the Module B expansion will be initiated. This module will investigate the safety and initial antitumor activity of the CLN-619 combination with pembrolizumab by enrolling patients with NSCLC, endometrial cancer, and up to three other tumor types. The Module B expansion will only be initiated after six patients have been enrolled at the same dose level and the safety of that dose level has been declared in accordance with the 3+3 design decision rule.

[0445] • Expanded B1: CLN-619 + pembrolizumab treatment for NSCLC patients (N=16)

[0446] • Expanded B2: CLN-619 + pembrolizumab treatment for endometrial cancer patients (N=16)

[0447] • During the dose escalation phase of the study, up to three additional disease-specific cohorts may be declared based on evidence of clinical benefit (objective response or stable disease throughout at least six treatment cycles) for that tumor type. Alternatively, these additional tumor-specific expansion cohorts may be triggered by considering the relationship between hematologic and tumor-based biomarker data and objective response.

[0448] • Expand the B1 and other disease-specific expanded cohorts: If 2 out of 16 patients experience clinical benefit (defined as an objective response or stable disease throughout at least 6 treatment cycles), the cohort may be further expanded to recruit up to 40 patients in total (including those treated during the dose escalation phase of the study) to further characterize the safety and preliminary antitumor activity of CLN-619 monotherapy in this cohort.

[0449] • Expanding B2: Patients will be alternately assigned to receive one of two dose levels of CLN-619 in combination with pembrolizumab. If 2 out of 16 patients experience clinical benefit (defined as an objective response or stable disease throughout at least 6 treatment cycles), this cohort may be further recruited to a total of up to 40 patients.

[0450] Module C CLN-619 + Chemotherapy Combination Therapy, Incremental and Expanded

[0451] Module C: Dose Escalation: Each chemotherapy combination cohort will be studied at 3 mg / kg and 10 mg / kg doses. Patients will be recruited according to a 3+3 design with 3 mg / kg CLN-619 in combination with a standard dose of chemotherapy. For each tumor type, the CLN-619+chemotherapy combination will start at 3 mg / kg (n=3-6) and escalate to 10 mg / kg (n=3-6) once safety is confirmed. If a DLT occurs at 10 mg / kg, the lower dose will be selected, and the safety process will be repeated. Safety data for each dose level tested for each tumor type in combination with chemotherapy will be reviewed to determine the optimal dose for recruiting the remaining patients to dose escalation, up to a maximum of 40 patients in total.

[0452] Module C Disease-Specific Dosage Expansion: Each cohort will initially recruit 16 patients. If 2 out of the 16 patients experience clinical benefit (defined as an objective response or stable disease throughout at least 6 treatment cycles), the cohort may be further expanded to recruit up to 40 patients in total to further characterize the safety and preliminary antitumor activity of the CLN-619 and chemotherapy combination in this cohort.

[0453] • Group C1: CLN 619 + carboplatin + pemetrexed for the treatment of relapsed EGFRm NSCLC

[0454] • Group C2: CLN 619 + Carboplatin + Paclitaxel for the treatment of recurrent endometrial cancer

[0455] • Group C3: CLN 619 + Paclitaxel for the treatment of platinum-resistant epithelial ovarian cancer

[0456] • Each treatment cycle will consist of 21 days or 3 weeks (q21d or q3w).

[0457] Table 5: C1: Relapsed EGFRm NSCLC

[0458]

[0459] Table 6: C2: Recurrent endometrial cancer

[0460]

[0461] Table 7: C3: Platinum-resistant epithelial ovarian cancer

[0462]

[0463] *Weekly dosage may be determined by the researcher as a matter of routine.

[0464] If DLT occurs at 10 mg / kg, a lower dose will be selected and the safety procedure will be repeated.

[0465] Module D Loading Dose Group

[0466] This module will investigate the safety and antitumor activity of CLN-619 monotherapy. A loading dose of 10 mg / kg will be administered weekly during the first cycle (QWx3). This will then be followed by dosing at 10 mg / kg Q3W in the second and subsequent cycles (following the standard dosing schedule).

[0467] Selection of research group

[0468] Planned number of patients

[0469] The total number of patients recruited across all modules will reach approximately 640, not including those requiring replacement. For Module A, monotherapy dose escalation, 45 patients with advanced solid tumors have been recruited. Up to 240 patients with specific tumor types will be recruited to the Module A monotherapy expansion group. If initiated, 25 patients have been recruited to the Module B combination therapy dose escalation group, up to 200 patients will be recruited to Module B, and up to 120 patients will be recruited to the Module C CLN-619 + chemotherapy combination therapy, escalation, and expansion groups. For the Module D monotherapy loading dose group, up to 10 patients will be recruited.

[0470] Inclusion criteria

[0471] Patients must meet all of the following inclusion criteria to be eligible to participate in the study.

[0472] 1) Males or females aged 18 or older.

[0473] 2) Willing and able to provide written informed consent and comply with the requirements of the agreement; prior to performing any procedures related to the agreement (including screening and evaluation), written informed consent and authorization as required by local regulations must be obtained from the patient.

[0474] 3) Module A monotherapy dose-escalation group and Module B combination therapy dose-escalation group: histologically or cytologically confirmed metastatic or locally advanced, unresectable solid tumors. For Module B, the tumor type is listed as the approved indication based on the current prescribing information for pembrolizumab.

[0475] 4) Expanding Module A Group:

[0476] a) Expanded A1: Metastatic or locally advanced, unresectable NSCLC confirmed by histology or cytology;

[0477] b) Expanded A2: Metastatic or locally advanced, unresectable cervical cancer confirmed by histology or cytology.

[0478] c) Expanded A3 and A4: Metastatic or locally advanced, unresectable endometrial cancer confirmed by histology or cytology.

[0479] d) Based on clinical, pharmacodynamic, or biomarker data from the study, the eligibility of disease-specific expanded groups can be further refined by histological subtypes, molecular characteristics, or exposure to prior therapies.

[0480] 5) Expanding Module B group:

[0481] a) Expanded B1: Metastatic or locally advanced, unresectable NSCLC confirmed by histology or cytology.

[0482] b) Expanded B2: Metastatic or locally advanced, unresectable endometrial cancer confirmed by histology or cytology.

[0483] c) Based on clinical, pharmacodynamic, or biomarker data from the study, the eligibility of the disease-specific expanded cohort can be further refined by histological subtypes, molecular characteristics, or exposure to prior therapy.

[0484] 6) Module C CLN-619 + chemotherapy combination therapy, incremental and expanded groups

[0485] a) Expanded C1: Histologically or cytologically confirmed recurrent NSCLC.

[0486] b) Extended C2: Histologically or cytologically confirmed recurrent endometrial cancer.

[0487] c) Expanded C3: Histologically or cytologically confirmed recurrent platinum-resistant epithelial ovarian cancer.

[0488] d) Based on clinical, pharmacodynamic, or biomarker data from the study, the eligibility of disease-specific expanded groups can be further refined by histological subtypes, molecular characteristics, or exposure to prior therapies.

[0489] 7) Module D Single Therapy Loading Dose 10 mg Group: Patients with recurrent epithelial ovarian cancer, breast cancer, and gastrointestinal cancer (esophageal cancer, gastric cancer, colorectal cancer).

[0490] 8) Previous medical history is as follows:

[0491] a) The patient should receive any other approved standard therapy available to the patient, unless the therapy is contraindicated, intolerable, or refused by the patient. If the patient refuses such therapy, proof that the patient was informed and refused should be recorded in the medical records.

[0492] 9) At baseline, the patient must have one or more measurable lesions that meet the RECIST v1.1 criteria and satisfy the following conditions:

[0493] a) Non-lymph node lesions with the longest single-dimensional measurement value ≥10mm or lymph node lesions with the shortest single-dimensional measurement value ≥15mm;

[0494] b) For lesions that have already received previous local treatments (such as radiotherapy or ablation), if the progression has been confirmed according to RECIST v1.1 prior to recruitment and the longest single-dimensional measurement is ≥10 mm, they can also be considered as measurable target lesions.

[0495] 10) According to the Eastern Cooperative Oncology Group (ECOG) physical fitness scale, the physical fitness status is 0 or 1.

[0496] 11) The estimated lifespan is 12 weeks or longer.

[0497] 12) Prior palliative radiotherapy must have been completed 14 days before C1D1 administration.

[0498] 13) According to the National Cancer Institute (NCI) Common Terminology Standard for Adverse Events (CTCAE) v5.0, toxicities associated with the previously investigated therapy should have regressed to Grade 1 or lower, except for alopecia. Peripheral neuropathy should be clinically stable or improved, and of Grade 2 or lower severity. Patients with long-term but stable Grade 2 toxicities may be permitted to be recruited upon agreement between the investigator and the sponsor.

[0499] 14) Possesses sufficiently good liver and kidney function, and hematological parameters are within the normal range, specifically defined as follows:

[0500] a) Total bilirubin ≤ 1.5x ULN. This does not apply to patients with a confirmed diagnosis of Gilbert's syndrome, in whom total bilirubin must be less than 3.0 mg / dL and conjugated bilirubin must be less than 0.5 mg / dL;

[0501] b) For patients with liver metastases, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) ≤2.5x ULN or ≤5x ULN;

[0502] c) Creatinine clearance (CrCl) ≥ 45 mL / min, if measured or estimated using the Cockcroft-Gault formula;

[0503] d) Hemoglobin ≥8 g / dL without blood transfusion for at least two weeks prior to C1D1 administration;

[0504] e) Absolute neutrophil count ≥1500 cells / mm² without growth factor support 3 Figexin for three days, polyethylene glycol-modified Figexin for 14 days;

[0505] f) Platelet count ≥75,000 cells / mm 3 .

[0506] 15) Patients in the dose escalation groups of Modules A and B, as well as Module D, must have archived tissue available for biomarker analysis. Samples should preferably be obtained after the most recent therapy. If archived tissue is unavailable (e.g., all tumor masses have been used up), a fresh biopsy is required. If a biopsy cannot be performed at the researcher's discretion due to unacceptable clinical risk, the sponsor's medical monitor must be contacted for recruitment approval.

[0507] a) Patients participating in the dose level group expansion of Module A and Module D must agree to provide a fresh biopsy at screening and an in-treatment biopsy on day 8 of cycle 2.

[0508] (b) Patients in the expanded cohorts of Modules A, B, and C must consent to a fresh pre-treatment biopsy and an in-treatment biopsy on day 8 of cycle 2. If the required biopsy cannot be obtained at the investigator's discretion with acceptable clinical risk, the sponsor's medical monitor must be contacted for recruitment approval. Matched biopsies are recommended if clinically feasible for the Modules A1 and B1 NSCLC cohorts.

[0509] Exclusion criteria

[0510] A patient will be excluded if they meet any of the following exclusion criteria:

[0511] 1) Currently or previously involved in interventional studies and received the study drug within 28 days (or five half-lives, whichever is longer) of C1D1 administration.

[0512] 2) Patients with a second malignancy (other than well-treated non-melanoma skin cancer, ductal carcinoma in situ, superficial bladder cancer, prostate cancer, or cervical cancer in situ) will be excluded unless they have achieved complete remission three years prior to study entry and do not require or are not expected to require additional therapy during study participation.

[0513] 3) Patients with any active autoimmune disease or a history of known or suspected autoimmune disease, or a history of a syndrome requiring systemic corticosteroid or immunosuppressive therapy, except for patients with vitiligo, relapsed childhood asthma / atopic disease, or autoimmune thyroid disease receiving stable thyroid hormone supplementation.

[0514] 4) Severe, uncontrolled medical conditions that impair a patient's ability to receive the prescribed therapy, or whose control may be jeopardized by complications of that therapy. These criteria include, but are not limited to, the following:

[0515] a) Uncontrolled airway hyperresponsiveness;

[0516] b) Type 1 diabetes. Individuals with type 2 diabetes are permitted to participate if their blood glucose control is stable, based on the researchers' assessment.

[0517] c) Uncontrolled, clinically significant lung disease;

[0518] d) Requires supplemental oxygen to maintain a pulse rate ox > 93%;

[0519] e) Symptomatic congestive heart failure as assessed by researchers or a recorded cardiac ejection fraction of less than 45%;

[0520] f) Patients with a history of prior anthracycline chemotherapy or a history of ventricular dysfunction have an ejection fraction <45%. Patients with a history of ventricular dysfunction or anthracycline therapy should undergo echocardiography to assess baseline cardiac function;

[0521] g) History of unstable angina or myocardial infarction within six months of C1D1 administration;

[0522] h) Unstable cardiac arrhythmias;

[0523] i) History of ventricular arrhythmias;

[0524] j) Uncontrolled hypertension: Patients with a sustained systolic blood pressure reading greater than 150 or a diastolic blood pressure greater than 100 should have a certificate from their attending physician proving that the finding is inconsistent with uncontrolled hypertension.

[0525] k) History of stroke or cerebral hemorrhage within one year of C1D1 administration;

[0526] l) Poorly controlled epilepsy;

[0527] m) Had active diverticulitis within one year prior to C1D1 administration;

[0528] n) Having undergone major surgery within three months of C1D1 administration, or major surgery with unresolved complications, may interfere with the study treatment.

[0529] 5) Treat acute infections with systemic antiviral, antibacterial, or antifungal agents within ≤7 days of C1D1 administration.

[0530] 6) History of HIV-1 / 2 primary immunodeficiency disease (such as human immunodeficiency virus (HIV)) or positive test result.

[0531] 7) Being diagnosed with hepatitis B (positive hepatitis B surface antigen [HBsAg] or hepatitis B core Ab test) or hepatitis C virus (HCV) infection (positive HCV antibody and / or HCV ribonucleic acid [RNA] test in serum) under any of the following conditions:

[0532] a) Active hepatitis B or hepatitis C disease and having received antiretroviral therapy within 4 weeks.

[0533] b) Blood hepatitis B DNA or HCV RNA can be detected.

[0534] 8) Previous organ allogeneic transplantation or allogeneic hematopoietic transplantation.

[0535] 9) History of any of the following events that occurred with prior treatment with checkpoint inhibitor immunotherapy: grade 3 or higher neurotoxicity, ocular toxicity, pneumonia, myocarditis, or colitis; liver dysfunction meeting laboratory criteria for Hy's law.

[0536] 10) Symptomatic, uncontrolled brain metastases, known or suspected leptomeningeal metastases, and / or carcinomatous meningitis. Patients with brain metastases identified at screening may be rescreened after receiving appropriate treatment (e.g., surgery and / or radiation therapy). Patients with brain metastases who have received treatment should maintain neurological stability for 28 days prior to completion of treatment and recruitment, and maintain a stable steroid dosing regimen (prednisone <10 mg or equivalent) for 14 days prior to C1D1 administration.

[0537] 11) Treatment with a non-tumor vaccine to control an infectious disease (i.e., HPV vaccine) within 28 days of C1D1. Inactivated seasonal influenza vaccines can be given to patients without restriction before the start of treatment and during investigational therapy. Live virus-containing influenza vaccines, or other clinically indicated infectious disease vaccines (i.e., pneumococcal vaccines, varicella vaccines), may be permitted, but must be discussed in advance with the sponsor's medical monitor, and an investigational drug washout period may be required before and / or after vaccine administration. Covid-19 vaccines may be administered according to institutional policy.

[0538] 12) Active SARS-CoV-2 infection, including a history of a positive SARS-CoV-2 test but no subsequent negative test result, patients with pending results, or patients with suspected active infection based on clinical characteristics. SARS-CoV-2 vaccination is permitted during treatment.

[0539] 13) Received immunosuppressive drugs within 28 days of C1D1 administration, including but not limited to cellcept, methotrexate, infliximab, anakinra, tocilizumab, cyclosporine, or corticosteroids (≥10 mg / day prednisone or equivalent).

[0540] 14) Women of fertility (FOCBP) who meet the following criteria: are pregnant or breastfeeding, plan to become pregnant within 120 days of the last dose of study drug, or refuse to use an acceptable method of contraception during study treatment and within 120 days of the last dose of study drug.

[0541] a) A woman of fertility is defined as:

[0542] i) Not sterilized by surgery, namely bilateral tubal ligation, bilateral oophorectomy, or total hysterectomy;

[0543] ii) Non-postmenopausal, defined as amenorrhea for ≥2 years without other medical cause.

[0544] Note: For women who have been amenorrhea for less than two years and have not undergone surgical sterilization (i.e., tubal ligation, bilateral oophorectomy, or total hysterectomy), they are considered infertile only if their follicle-stimulating hormone (FSH) levels are within the postmenopausal range.

[0545] 15) Male patients who: plan to conceive or donate sperm within 120 days of the last dose of study drug or within 5 half-lives of CLN-619 (whichever is later), or whose partner is FOCBP, and who refuse to use acceptable methods of contraception during study treatment and within 120 days of the last dose of study drug or within 5 half-lives of CLN-619 (whichever is later).

[0546] 16) The QT interval for heart rate correction using the Fridericia formula (QTcF) is ≥ 500 milliseconds.

[0547] 17) The patient has a history of drug-related allergic reactions to any component of CLN-619 (module A and module B patients) or pembrolizumab (module B patients only). A history of grade 4 allergic reactions to any monoclonal antibody therapy.

[0548] 18) Known alcoholism or drug abuse.

[0549] 19) Unable to comply with the agreement and / or unwilling or unable to conduct follow-up assessments.

[0550] 20) Patients who have lost their capacity for civil conduct or are involuntarily imprisoned.

[0551] 21) Patients deemed unsuitable for participation based on the researcher's judgment.

[0552] 22) Use any of the following treatments:

[0553] a) Received systemic anticancer therapy within 14 days prior to the first dose of the study drug C1D1.

[0554] b) Received immunotherapy ≤28 days prior to the first dose of the study drug C1D1.

[0555] c) Prior to the first dose of the C1D1 study drug, radiation therapy must have been administered for <28 days and palliative radiation for ≤14 days. If radiation therapy is received, the lesion must have shown significant progression before it qualifies as a target lesion for evaluation.

[0556] d) Underwent major surgery (excluding vascular access placement) within ≤28 days of the first dose of the study drug C1D1.

[0557] Diet and dietary restrictions

[0558] Patients should maintain a normal diet unless adjustments are needed to control adverse events such as diarrhea, nausea, or vomiting.

[0559] Criteria for Treatment Discontinuation

[0560] Patients who have discontinued treatment should be encouraged to return for safety follow-up and survival follow-up. If a patient withdraws from treatment and follow-up, no further evaluation or additional data should be collected.

[0561] The reasons for treatment discontinuation include, but are not limited to, the following:

[0562] • Complete 34 cycles of therapy;

[0563] • Disease progression or a requirement for cancer treatment not specified in the agreement;

[0564] • When a patient's condition develops complications, general or specific changes, and the researchers believe that further treatment is not in the best interest of the patient;

[0565] • die

[0566] • DLT or uncontrollable adverse events related to the therapy;

[0567] • Withdrawal of treatment consent;

[0568] • Serious breach of the agreement or non-compliance with the agreement;

[0569] • The sponsor terminated the study;

[0570] For patients who meet the criteria for treatment discontinuation but feel that they have received significant clinical benefit from continued treatment with CLN-619, pembrolizumab, or a combination of CLN-619 and pembrolizumab, or a combination of CLN-619 and chemotherapy, further investigation of the therapy may be considered on a case-by-case basis.

[0571] Research termination criteria (EOT)

[0572] Patients should be encouraged to complete all study evaluations. However, patients may withdraw their consent to participate in this study at any time for any reason without penalty or loss of their entitlements.

[0573] Patients will withdraw from the study (survival follow-up) in the following circumstances:

[0574] • Complete the follow-up period;

[0575] • Withdrawal of follow-up consent;

[0576] • Patients lost to follow-up;

[0577] • die;

[0578] • The sponsor, Institutional Review Board (IRB) / Ethics Committee (EC), regulatory agency or other governing body terminates the study.

[0579] Definition of dose-limiting toxicity (DLT)

[0580] Throughout the study, treatment-related adverse events (DLTs) will be defined by the use of CLN-619 alone, in combination with pembrolizumab, or in combination with chemotherapy as an investigational drug-related adverse event (Ae) that is possibly or definitely treatment-related. DLTs will be defined separately for hematologic events, non-hepatic non-hematologic events, and hepatic non-hematologic events, as described below. The severity of all events will be graded according to CTCAEv5.0. For patients experiencing grade 3 or higher adverse events, treatment with the investigational drug should generally be suspended until the event is managed and resolved, and the relationship between the event and the administration of the investigational drug should be assessed.

[0581] Hematological dose-limiting toxicity (DLT):

[0582] • Grade 4 neutropenia lasting ≥7 days;

[0583] • Grade 4 febrile neutropenia;

[0584] • Grade 3 febrile neutropenia lasting >48 hours or with objective evidence of clinical infection;

[0585] • Grade 4 thrombocytopenia or Grade 3 or higher thrombocytopenia with clinically significant bleeding.

[0586] Non-hepatic, non-hematologic dose-limiting toxicities (DLT):

[0587] Grade 3 or higher non-hematologic, non-hepatic arterial lesions (Ae) will be considered DLT, except in the following cases:

[0588] • Grade 3 or 4 electrolyte abnormality, without clinical complications, and responding to medical intervention within 96 hours;

[0589] • Level 3 fatigue lasts for less than 72 hours;

[0590] • Level 3 fever;

[0591] • Grade 3 headache, responding to medical intervention within 24 hours;

[0592] • Grade 3 nausea, vomiting, or diarrhea, with a response to medical intervention within 72 hours;

[0593] • An elevation of grade 3 or 4 amylase or lipase in the absence of radiological or clinical signs and symptoms suggestive of pancreatitis;

[0594] • Grade 3 hypertension, without clinical complications, and responding to medical intervention within 24 hours;

[0595] Hepatic, non-hematologic dose-limiting toxicities (DLT):

[0596] Elevated transaminase levels

[0597] • Grade 3 or higher AST or ALT elevation >10x ULN, regardless of duration;

[0598] • A grade 3 AST or ALT elevation of ≥ 5x ULN and ≤ 10x ULN, which fails to regress to grade 2 or below within seven days and fails to regress to grade 1 or below within 14 days.

[0599] Elevated bilirubin

[0600] • Grade 3 or higher total bilirubin elevation >5x ULN, regardless of duration;

[0601] • Grade 3 bilirubin elevation ≥ 3x ULN and ≤ 5x ULN, which fails to decline to Grade 2 or below within seven days and fails to decline to Grade 1 or below within 14 days.

[0602] Adverse events that meet the criteria of Hy's Law will also be considered DLT:

[0603] • AST or ALT ≥ 3x ULN, and

[0604] • Total bilirubin ≥ 2 x ULN (no cholestasis), and

[0605] • Lack of provable alternative causes.

[0606] Grade 2 events that are persistent, clinically significant, and require continuous dose interruption in patients can also be considered DLT, depending on the specific circumstances.

[0607] Study the stopping rules

[0608] If one Grade 5 event or two Grade 4 events are considered potentially related to CLN-619 therapy, the study accumulation will be paused. Grade 4 laboratory abnormalities that regress to Grade 2 or below within 96 hours will not be included.

[0609] Decision rules for accelerated titration groups in Module A

[0610] Accelerated titration will switch to the standard 3+3 design at the 3.0 mg / kg dose level unless toxicity at lower dose levels necessitates starting from a lower dose level.

[0611] After completing the accelerated titration group for a single patient, if:

[0612] • No CLN-619-related grade ≥ 2 Ae occurred during the first cycle; dose escalation may lead to the next accelerated titration group.

[0613] • Patients experience any CLN-619-related grade ≥ 2Ae during cycle 1; convert the current cohort to a 3+3 design and recruit two to five additional patients.

[0614] • The patient underwent DLT; the design was switched to 3+3 and two to five more patients were recruited.

[0615] Decision rules for the 3+3 dose escalation group in Module A

[0616] After completing the 3+3 group (minimum three, maximum six patients), if:

[0617] • None of the three patients experienced DLT; dose escalation can proceed to the next cohort.

[0618] • One of the three patients will have experienced DLT; the other three patients will be recruited into the group.

[0619] • ≤ 1 out of 6 patients experienced DLT; dose escalation can proceed to the next cohort.

[0620] • ≥ 2 patients have experienced DLT; MTD has been exceeded, and further recruitment at this dose level will be discontinued.

[0621] ○ Then, the lower dose group, including intermediate doses, can be further explored until the MTD is determined.

[0622] Decision rules for the 3+3 dose escalation group in Module B

[0623] After completing the dose escalation level group (minimum three, maximum six patients), if:

[0624] • None of the three patients experienced DLT; dose escalation can proceed to the next dose level group.

[0625] • One of the three patients will have experienced DLT; the other three patients will be recruited to the current dose level group.

[0626] • ≤ 1 out of 6 patients experienced DLT; dose escalation can proceed to the next dose level group as long as the safety of CLN-619 at the next escalation dose level is determined by SRC during the dose escalation of the monotherapy in Module A of the study.

[0627] • ≥ 2 patients have experienced DLT; MTD has been exceeded, and further recruitment at this dose level will be discontinued.

[0628] ○ According to the 3+3 decision rule mentioned above, a lower dose "minus one" group can be recruited.

[0629] - If there are ≥ 2 patients in the "Minus 1" cohort who have experienced DLT, the sponsor may proceed with the "Minus 2" cohort at a lower dose, following the 3+3 decision rule described above.

[0630] If there are ≥ 2 patients in the "minus 2" cohort who have experienced DLT, the sponsor will not proceed with the expansion of the Module B cohort.

[0631] Decision rules for dose escalation in Module C (3 mg / kg and 10 mg / kg)

[0632] Determine the safety at a dose level of 3 mg / kg

[0633] • None of the three patients experienced DLT at the 3 mg / kg dose level; dose escalation was possible up to the 10 mg / kg dose level.

[0634] • At a dose level of 3 mg / kg, 1 out of 3 patients experienced DLT; three more patients will be recruited.

[0635] • ≤ 1 out of 6 patients experienced DLT; dose escalation can be performed up to a dose level of 10 mg / kg.

[0636] • At a dose level of 3 mg / kg, ≥ 2 patients experienced DLT; this has exceeded MTD, and further recruitment at 3 mg / kg will be discontinued.

[0637] Determine the safety at a dose level of 10 mg / kg

[0638] • None of the three patients experienced DLT at the 10 mg / kg dose level, and dose escalation can be performed at 10 mg / kg.

[0639] • At the 10 mg / kg dose level, 1 out of 3 patients will experience DLT; three more patients will be recruited at the 10 mg / kg dose level.

[0640] • At a dose level of 10 mg / kg, ≤ 1 out of 6 patients experienced DLT; dose escalation can be performed at 10 mg / kg.

[0641] • If ≥ 2 patients experience DLT at 10 mg / kg, exceeding the MTD, further recruitment at 10 mg / kg will be discontinued. Data for each dose level tested for each tumor type in combination with chemotherapy will be reviewed to determine the dose for recruiting the remaining patients to dose expansion (up to 40 patients in total).

[0642] Decision rules for dose escalation in module D

[0643] • To assess the safety of the dosing regimen (10 mg / kg, first cycle, Q1W and second cycle and thereafter, Q3W), 3–6 patients will be initially recruited. If fewer than 2 dose-limiting toxicities (DLTs) are observed, additional patients (up to 10) may be recruited to collect additional PK and biomarker data to establish this dosing regimen; if none of the 3 patients experienced DLTs, additional patients may be recruited.

[0644] • One of the three patients has experienced DLT; three more patients will be recruited.

[0645] • If ≤ 1 out of 6 patients has experienced DLT, additional patients may be recruited. If ≥ 2 patients have experienced DLT, the MTD has been exceeded, and further recruitment according to this dosing schedule will be discontinued.

[0646] Dosage scalation decision rules

[0647] During the study, if >33% of patients experience Ae that meets the DLT criteria during the first three-week cycle, or if a safety finding suggesting clinically significant cumulative toxicity is observed, the SRC may be temporarily convened to review data from the expanded cohort and make recommendations on continued dosing for patients.

[0648] Determination of the maximum bioeffective dose

[0649] Both Module A (monotherapy) and Module B (combination therapy) dose escalation are designed to determine the MTD, MAD (if MTD is not defined), and / or MBED of CLN-619 or CLN-619 in combination with pembrolizumab, and to inform the selection of RP2D for further studies in their respective congregation expansions. During this phase, in addition to safety and efficacy data, PK and exploratory biomarker data will be collected and analyzed to assess whether doses below the MTD or MAD have a more favorable clinical profile.

[0650] Timing and frequency of dose administration

[0651] All infusions will be administered in a monitored environment with immediate access to trained personnel and adequate equipment and medication to manage potentially serious or life-threatening reactions. In dose escalation (Modules A and B), CLN-619 will be administered as a 60-minute IV infusion (+10 / - 5 minutes) on Day 1 of each three-week treatment cycle. Baseline body weight will be used to calculate the CLN-619 dose. Baseline body weight is the weight obtained at screening. If there is a change in weight of + / - 10%, the dose will be adjusted.

[0652] In Module B, pembrolizumab 200 mg will be administered via 30-minute IV infusion (+10 / -5 minutes) on Day 1 of each three-week treatment cycle. For more information on pembrolizumab administration, please refer to the Pharmacy Manual. In Module B of this study, when CLN-619 is administered in combination with pembrolizumab, a prophylactic agent (including dexamethasone) will be administered first, followed by pembrolizumab. CLN-619 will be administered immediately after the completion of the pembrolizumab infusion (+30 minutes). Administration of the combination therapy may be delayed if an infusion-related reaction occurs during pembrolizumab administration.

[0653] In Module C, standard care chemotherapy should be administered on Day 1 of each 3-week treatment cycle prior to CLN-619 administration, according to prescribing information or institutional policy. In Module D, CLN-619 (10 mg / kg) will be administered over 120 minutes on Day 1 of the first cycle, which is the first dose of the three weekly doses in the first cycle. For all subsequent weekly doses (C1D8 and C1D15) and cycles (Cycle 2 and thereafter), the infusion may be administered over 60 minutes if the patient does not experience an IRR.

[0654] Prior to CLN-619 administration, a pre-infusion of medication, including the required corticosteroid, will be administered. CLN-619 infusion will be administered via IV infusion over 120 (+10 / -5) minutes on Day 1 of Cycle 1. Baseline body weight will be used to calculate the CLN-619 dose. If no infusion-related response occurs during the first treatment cycle, all subsequent three-week cycles will be administered via IV infusion over 60 (+10 / -5) minutes. If an infusion-related response occurs during the Day 1 visit of Cycle 1, CLN-619 administration on Day 1 of Cycle 2 will be administered via IV infusion over 120 (+10 / -5) minutes, and subsequent cycles will be administered via IV infusion over 60 (+10 / -5) minutes. Figure 26A and 26B If an IRR occurs after a C2D1 visit, the clinical site should contact the sponsor for guidance on future infusions.

[0655] Predrugs used for the prevention of infusion-related reactions

[0656] For patients to be treated with CLN-619 alone, in combination with pembrolizumab, or in combination with chemotherapy, prophylactic medication for infusion-related reaction prophylaxis using corticosteroids, antihistamines, and / or antipyretics is mandatory. Suitable corticosteroids include dexamethasone; suitable antihistamines include diphenhydramine and famotidine; suitable antipyretics include paracetamol, acetaminophen, and ibuprofen. Folic acid supplementation may also be administered.

[0657] Classification and management of infusion-related reactions

[0658] Infusion reactions may manifest as fever, chills, shivering, headache, rash, itching, joint pain, low or high blood pressure, bronchospasm, or other symptoms.

[0659] Table 8 below provides further treatment recommendations, which may be modified as appropriate based on local treatment standards and guidelines.

[0660] Table 8: Management of Infusion-Related Reactions

[0661]

[0662]

[0663] Tumor imaging and assessment of disease response according to RECIST v 1.1

[0664] Researchers will read the images locally and conduct efficacy response assessments based on their evaluation. Enhanced CT is strongly preferred for tumor imaging. Contrast-enhanced MRI may be used for the chest, abdomen, and pelvis when CT with iodinated contrast agents is contraindicated or mandated by local practice. MRI is strongly preferred for brain imaging when clinically indicated. Enhanced CT is an acceptable alternative if MRI is medically contraindicated.

[0665] Throughout the study, the same scanner and contrast agent should be used on the same patient to optimize the reproducibility of assessments of existing and new tumor burden and improve the accuracy of imaging-based assessments of response or progression.

[0666] Any patient who completes 34 treatment cycles will undergo tumor imaging at the safety follow-up. Patients who have undergone tumor imaging ≤12 weeks prior to this visit and discontinue treatment before 34 cycles may not require tumor imaging at this visit.

[0667] RECIST assessment of the disease

[0668] In this study, RECIST v1.1 will be used as the primary metric for assessing tumor response and for making treatment decisions.

[0669] For patients with an objective response, a confirmatory scan should be performed at least four weeks after the initial scan in which the response was detected, or at the next scan required by the protocol.

[0670] Clinical stability is defined as follows:

[0671] • Lack of symptoms and signs that indicate significant clinical progression of the disease.

[0672] • No intensive management is required, including enhanced analgesia, radiation or other palliative care.

[0673] • For more information about RECIST, see https: / / recist.eortc.org / recist-1-1-2 / .

[0674] Pharmacokinetics

[0675] Blood samples will be collected to assess serum concentrations of CLN-619 and pembrolizumab (Module B only) and to determine relevant pharmacokinetic parameters.

[0676] Immunogenicity

[0677] Blood samples will be collected to assess immunogenicity parameters, including human anti-drug (CLN-619 and pembrolizumab (Module B only)) antibody titers (ADA).

[0678] Definition of AE

[0679] An AE (Adverse Event) refers to any adverse medical event or exacerbation of a pre-existing medical condition that occurs in a clinical trial participant who has received a medical product and is not necessarily causally related to the treatment.

[0680] Therefore, AE can be:

[0681] • Any adverse or unexpected clinical signs (e.g., including abnormal laboratory findings).

[0682] • Any symptoms, illnesses, or injuries that are related to the use of the study product, whether or not they are related to the study product.

[0683] Ae will not include:

[0684] • Medical or surgical procedures, such as endoscopy, tooth extraction, or intravenous infusion (although the underlying condition that leads to the procedure may be an AE).

[0685] • Pre-existing diseases or conditions that were present at the start of the study will not worsen during the study.

[0686] • No adverse medical events occurred (e.g., hospitalization due to elective cosmetic surgery or social admission).

[0687] • Researchers identify laboratory test results that are not clinically significant.

[0688] Adverse events can be spontaneously reported or triggered during open-ended inquiries, examinations, or evaluations of patients. (To prevent reporting bias, patients should not be asked about the specific occurrence of one or more adverse events.)

[0689] Chronic diseases present before the start of the study should be recorded in the medical history section of the eCRF and reported as Ae only if the frequency or severity of the disease increases during the study.

[0690] Natural progression or worsening of malignant tumors in the study will be recorded as part of the efficacy assessment and should not be recorded as an AE / SAE. Death due to disease progression will be recorded as part of the efficacy assessment and will not be considered an SAE.

[0691] If the signs and symptoms of disease progression are clearly associated with recurrence or with anticipated changes in baseline malignancy, they should not be reported as Ae / SAE. Ae / SAE should only be reported if these signs and symptoms meet the following criteria (at the researcher's discretion):

[0692] • Determined by researchers to be an abnormally severe or accelerating malignant tumor, or

[0693] • If researchers believe that the worsening of signs and symptoms of malignant tumors is directly caused by IMP.

[0694] If it is uncertain whether an AE is caused solely by a malignant tumor in the study, it should be reported as an AE / SAE as appropriate.

[0695] Qualified physicians, researchers, and any designated personnel are responsible for detecting, evaluating, recording, and reporting events that meet the definition of an AE or SAE, as well as other reportable safety incidents. Researchers remain responsible for following up on AEs, SAEs, and other reportable safety incidents to understand their outcomes.

[0696] Adverse event severity classification

[0697] The severity of adverse events (Ae) will be documented according to NCI CTCAE v5.0. This guideline provides a common language for describing severity levels, analyzing and interpreting data, assessing overall AE scores, and clarifying the clinical significance of all Ae. A copy of the current NCI CTCAE version can be downloaded from http: / / evs.nci.nih.gov / ftp1 / CTCAE / About.html.

[0698] For Ae not listed in NCI CTCAE, the following levels should be used:

[0699] • Level 1: Mild AE – asymptomatic or mild symptoms; clinical or diagnostic observation only; no intervention indicated.

[0700] • Level 2: Moderate AE – Indicates minimal, local or non-invasive intervention; age-appropriate activities of daily living are restricted.

[0701] • Level 3: Severe AE – Serious or medically significant but not immediately life-threatening; instructs hospitalization or prolongs hospitalization; causes disability; limits self-care activities in daily life.

[0702] • Level 4: Life-threatening or disabling AE – life-threatening consequences; requires emergency intervention.

[0703] • Level 5: Deaths related to AE

[0704] An AE assessed as severe is different from a serious AE. Severity is a category used to assess the intensity of an event, and both AEs and SAEs can be assessed as severe. An AE is defined as "severe" when it meets one of the predefined serious consequences below.

[0705] Sample size

[0706] No formal sample size estimation was performed.

[0707] Module A:

[0708] The number of patients in the dose escalation phase is estimated to be up to 50 in seven cohorts, including cohort expansions at all dose levels (45 patients have been recruited). It is estimated that up to 240 patients could be recruited to cohort expansion in module A (assuming full expansion). Overall, it is estimated that up to approximately 285 patients will be dosed in module A of this trial.

[0709] During dose escalation, patients who withdraw from treatment for reasons other than DLT during the three-week DLT monitoring period will be replaced.

[0710] Module B:

[0711] The number of patients requiring combination dose escalation is estimated to be up to 30 (assuming six patients receive up to five dose levels); 25 patients have been recruited. It is estimated that up to 200 patients can be recruited to the Module B expansion. Overall, it is estimated that up to approximately 225 patients will be dosed in Module B of this trial.

[0712] Module C:

[0713] The chemotherapy combination group will have a maximum of 40 patients per indication (40 each for NSCLC, endometrial cancer, and epithelial ovarian cancer). Overall, it is estimated that up to approximately 120 patients will be administered in module C of this trial.

[0714] Module D:

[0715] For the Module D single-therapy loading dose group, up to 10 patients will be recruited.

[0716] In the dose escalation group, patients who withdraw from the study for reasons other than DLT during the three-week DLT monitoring period will be replaced.

[0717] result:

[0718] Sixty-four patients received either the combination of CLN-619 and pembrolizumab (n=22) or monotherapy (n=42). Baseline characteristics are presented in Table 9 below.

[0719] Table 9: Baseline Characteristics

[0720]

[0721] CPI, checkpoint inhibitor; NSCLC, non-small cell lung cancer.

[0722] a Other tumor types in the cohort: gastric cancer (2 patients), esophageal cancer (1), head and neck cancer (1), skin cancer (1), and urothelial carcinoma (1); b Other tumor types in the single-therapy group: breast cancer (2), pancreatic cancer (2), sarcoma (2), adenoid cystic carcinoma (1), cecal cancer (1), duodenal cancer (1), head and neck cancer (1), renal cancer (1), leiomyosarcoma (1), mediastinal endometrial sarcoma (1), melanoma (1), parotid gland cancer (1), peritoneal mesothelioma (1), and thyroid cancer (1).

[0723] Of the 22 patients treated with CLN-619 plus pembrolizumab, 18 were assessable for response by 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), or admission to a hospice with acute kidney injury (n=1). Confirmed responses (all partial responses) were observed in three patients treated with CLN-619 in combination with pembrolizumab at doses ≥3 mg / kg. Responses were also observed in patients with tumor types that typically do not respond to checkpoint inhibitors (CPIs) alone.

[0724] The characteristics of the respondents are detailed in Table 10, and Figure 27 The treatment duration and clinical activities of the combined groups are described.

[0725] Table 10: Characteristics of Respondents

[0726]

[0727] Both monotherapy and combination therapy are effective against NSCLC, which is in Figure 28 The figure depicts the treatment duration and clinical activity in NSCLC patients. Objective responses and stable disease (SD) were observed in NSCLC patients with targetable oncogenic mutations in both the CLN-619 monotherapy and combination therapy groups.

[0728] Of the 42 patients treated with CLN-619 monotherapy, 29 received CLN-619 at a dose ≥1 mg / kg and were evaluable by RECIST. Six patients did not undergo post-baseline imaging for response evaluation due to clinical progression (n=3), end-of-life care (n=1), and withdrawal of consent (n=2). Seven patients received doses below the treatment dose (<1 mg / kg) or were not evaluable by RECIST. The clinical benefit rate (CBR) was 41.4% (1 complete response (CR), 2 partial responses, and 9 SD ≥18 weeks). Table 11 shows the characteristics of patients who responded or had an SD ≥18 weeks. Figure 29 The treatment duration and clinical activity of monotherapy groups ≥1 mg / kg are described.

[0729] Table 11: Characteristics of patients with a response or SD ≥ 18 weeks

[0730]

[0731] Safety: CLN-619 plus pembrolizumab combination therapy was well tolerated at doses ranging from 1 to 6 mg / kg and demonstrated clinical activity, including objective responses, in tumor types that are typically unresponsive to pembrolizumab. No adverse events (AEs) met the protocol-defined DLT criteria, and most TEAEs were grade 1 / 2 (see [link to relevant documentation]). Figure 30 Treatment-related adverse events (TRAEs) reported in ≥10% of patients were fatigue (combination therapy: 18.2%; monotherapy: 9.5%) and IRR (combination therapy: 18.2%; monotherapy: 28.6%). The only ≥Grade 3 treatment-related AE reported in >5% of patients in any group was elevated AST (combination therapy: 0%; monotherapy: 7.1%). TEAEs led to study discontinuation in 13.6% (3 / 22) of patients in the combination therapy group and 9.5% (4 / 42) of patients in the monotherapy group. IRR was a new-onset known AE of CLN-619, while IRR was the most frequently reported TRAE of CLN-619. Most IRRs were Grade 1 or 2 following administration of prophylactic prophylaxis, occurring on Day 1 of Cycle 1 and resolving rapidly. No treatment-related deaths occurred.

[0732] Conclusion: Objective responses were observed with CLN-619 + pembrolizumab in patients with tumor types that typically do not respond to pembrolizumab (e.g., NSCLC with ALKr and EGFRm). CLN-619 + pembrolizumab was well tolerated at doses ranging from 1 to 10 mg / kg. Long-term follow-up of patients treated with CLN-619 monotherapy confirmed good safety and durable clinical benefit, including objective responses in patients with various tumor types and those whose disease progressed after CPI therapy. Based on these findings, an expanded cohort is being recruited for endometrial cancer and NSCLC patients into the monotherapy and combination therapy groups. Table 12 shows the best confirmatory responses in RECIST-evaluable patients.

[0733] Table 12: Best Confirmatory Response (RECIST-evaluable patients)

[0734]

[0735] 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, stable disease.

[0736] CLN-619 also demonstrated single-agent activity, including objective responses to a variety of tumors.

[0737] Example 4. Conclusions on the application of CLN-619 (anti-MICA / B antibody) in solid tumors and its use in the treatment of multiple myelopathy Evidence of tumor

[0738] CLN-619 will also be studied in a phase 1 clinical trial (NCT06381141) in patients with relapsed / refractory multiple myeloma.

[0739] CLN-619 has demonstrated antitumor efficacy in the treatment of advanced and solid tumors, and has been used as a monotherapy and in combination therapy. Combination therapy is well tolerated.

[0740] In previous embodiments, 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 advanced solid tumors. These studies show that in vitro binding data collectively support the high specificity and potent binding of CLN-619 to MICA / MICB, thereby demonstrating broad reactivity and high affinity binding to representative alleles of MICA and typical alleles of MICB. CLN-619 exhibits multiple modes of action. By binding to MICA / MICB and preventing their hydrolysis from tumor cell surface proteins, CLN-619 enhances NKG2D-mediated immune cell activation through antibody-dependent cytotoxicity (ADCC) and antibody-dependent phagocytosis (ADCP) on tumor cells expressing MICA / MICB. Furthermore, in vivo studies have demonstrated that CLN-619 treatment produces potent antitumor activity in mouse models carrying human liver, lung, and multiple myeloma tumor xenografts expressing MICA / MICB, within a dose range of 0.1 mg / kg to 10 mg / kg. Antitumor efficacy was also observed in a lung cancer model at doses as low as 0.1 mg / kg.

[0741] NCT05117476, described in the above examples, recruited patients with advanced metastatic solid tumors in the CLN-619 (CLN-619-001) study. Multiple doses of CLN-619 (maximum dose 10 mg / kg) were tested, administered alone or in combination with pembrolizumab every three weeks. In this study, CLN-619 was tolerable in both monotherapy and combination therapy scenarios, with no dose-limiting toxicities observed during dose escalation, and no grade 3 treatment-related adverse events (TRAEs) occurred in patients receiving protocol-mandated progenitor drugs.

[0742] As of June 22, 2023, 61 patients received intravenous doses of CLN-619 at doses of 0.1, 0.3, 1, 3, 6, and 10 mg / kg according to a 21-day cycle protocol. Forty-three patients received CLN-619 monotherapy, with 10 receiving a dose of 3 mg / kg, 11 receiving a dose of 6 mg / kg, and 12 receiving a dose of 10 mg / kg. Eighteen patients received CLN-619 in combination with pembrolizumab at doses of 1, 3, 6, and 10 mg / kg, with 6 receiving a dose of 10 mg / kg. CLN-619 was found to be well-tolerated, with no adverse events (AEs) meeting the protocol-defined DLT criteria and no grade 4 transarterial adverse events (TRAEs).

[0743] Serious adverse events (SAEs) were monitored following administration of CLN-619 (monotherapy and in combination with pembrolizumab). Only one patient experienced an infusion-related reaction (IRR) event, laryngeal edema, at a dose level of 10.0 mg / kg, which was considered to be related to CLN-619. This patient was not using a prophylactic corticosteroid. One mild grade 3 rash was reported in the monotherapy group, which was considered to be possibly related. No serious adverse events related to CLN-619 were reported in the pembrolizumab combination therapy, but one case of acute kidney injury (AKI) SAE was reported, which was not related to CLN-619. To date, the most common treatment-related adverse events (TEAEs) in ≥ 5% of patients treated with CLN-619 monotherapy included IRR (23.3%), abdominal pain (18.6%), fatigue (18.6%), and fever (18.6%).

[0744] Most IRRs occurred during or immediately after the first dose of CLN-619; one IRR occurred after the sixth dose; and one patient had IRR after both the first and second doses. One event also occurred in a patient receiving combination therapy with pembrolizumab. All patients recovered without sequelae. Treatment recommendations include prophylactic therapy for IRR and a longer infusion time at the first administration of CLN-619. Monitoring for immune-related adverse events is also recommended, such as immune-mediated colitis, hepatitis, pneumonia, hypothyroidism, hyperthyroidism, adrenal insufficiency, hypophysitis, nephritis with renal dysfunction, myocarditis, and uveitis.

[0745] Evidence for using CLN-619 to treat multiple myeloma

[0746] Multiple myeloma (MM) is a plasma cell tumor that arises from the malignant transformation of antibody-producing plasma cells in the bone marrow. It is characterized by the increasing presence of abnormal monoclonal immunoglobulins (M proteins) in the serum and urine. Excessive production of M proteins can lead to fatal complications, including bone destruction, hypercalcemia, anemia, kidney damage, and an increased risk of infection.

[0747] Multiple myeloma (MM) accounts for approximately 10% to 18% of all hematologic malignancies. It is an older disease, with a median age of diagnosis of approximately 70 years, and is slightly more common in men than women. MM is 2 to 3 times more common in African American patients than in Caucasians, making it the most common hematologic malignancy in this population. Globally, there are approximately 160,000 new cases of MM each year. In the United States (US), there are approximately 34,500 new cases of MM and approximately 13,000 deaths from MM each year. The frequency of MM is geographically uneven, with the highest incidence in Europe, North America, and Australia / New Zealand. Despite some progress in recent years, multiple myeloma remains incurable, with the most common causes of death being disease progression, infection, and kidney failure. The median overall survival (OS) for MM patients is 2 to 10 years or more, with approximately 15% of patients dying within 2 years of diagnosis.

[0748] Treatment options for multiple myeloma (MM) include a variety of therapies, all aimed at controlling and eliminating the cells. These include proteasome inhibitors, immunomodulators, steroids, antibodies, BCMA-targeted therapies, and chemotherapy. Unfortunately, most MM patients experience recurrent relapses and require multiple lines of therapy. Relapsed MM cells often exhibit a more aggressive phenotype, leading to shorter response durations and reduced survival. Treatment at this stage typically requires a multifaceted approach to address the heterogeneous biological aspects of the disease and the full range of clinical complications associated with it. Optimal long-term disease control can be achieved by combining therapies with different mechanisms of action, particularly those targeting key components of the immune system. Therefore, new therapies for MM are urgently needed.

[0749] It has been well established that MM cells can detach MICA / MICB from their surface via proteolytic shedding to evade tumor cell surveillance. The loss of MICA / MICB from the cell surface allows tumor cells to evade immune cell recognition and destruction, accompanied by immune cell dysregulation due to NKG2D internalization on the immune cell surface, as well as impaired NK and CD8+ T cell function and expansion. High concentrations of detached MICA (sMICA) have been observed in the serum of patients from various tumor types and have been shown to be associated with poor survival, including in MM, where it is an independent prognostic factor for OS and PFS. MM patients with sMICA are known to have reduced NKG2D levels on NK and CD8+ T cells and decreased NK cell cytotoxicity. Furthermore, detached MICB (sMICB) has been shown to promote an immunosuppressive microenvironment by polarizing macrophages to the M2 phenotype and expanding myeloid-derived suppressor cells (MDSCs) in the tumor microenvironment (TME). Preclinical data have validated the role of the MICA / MICB-NKG2D axis in immune surveillance, as MICA overexpression in human tumor models led to delayed tumor growth and improved survival in mice. Furthermore, while NKG2D-deficient mice exhibit normal development, their tumor surveillance function is impaired in spontaneous malignant tumor models.

[0750] Immunomodulatory agents such as lenalidomide and pomalidomide can enhance the expression of MICA and CD155 by binding to cereblon and downregulating the transcription factors IKZF1, IKZF3, and IRF4 in MM cell lines and primary MM cells, thereby leading to the degranulation of interferon-γ in NK cells. Therefore, the upregulated MICA / MICB on myeloma cells can be stabilized by using anti-MICA / MICB stabilizing antibodies, thereby enhancing the elimination of myeloma cells by NK and T cells expressing NKG2D. Furthermore, 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 circulating and bone marrow-infiltrated NK and T cells. These changes included an increase in the number of NK and T cells and upregulation of activation receptors, including NKG2D. In this embodiment, the inventors suggest that combining CLN-619 with immunomodulators to increase the expression of MICA and MICB on malignant plasma cells and improve immune cell activation can further synergize these effects.

[0751] Some have proposed that therapeutic agents that effectively restore MICA / MICB expression on the surface of tumor cells are an effective way to reverse tumor-mediated immunosuppression in cancer patients with multiple myeloma.

[0752] While immunotherapy has provided improvements in overall survival and quality of life and dramatically altered the treatment prospects for patients with MM, most patients with advanced disease still experience disease progression during or after multiple double or triple therapy combinations, indicating an unmet need in patients with refractory disease. There is strong evidence to modulate innate and adaptive immune responses using CLN-619 monotherapy or in combination with pomalidomide and dexamethasone. This embodiment provides a planned combination regimen that uses FDA-approved standards of care for MM and has strong scientific and clinical data to support its use in combination with CLN-619 for the treatment of MM. The apoptotic activity induced by pomalidomide and dexamethasone can induce cellular stress and consequently increase the expression of NKG2D ligands on MM cells. CLN-619 further stabilizes these MICA and MICB ligands, which can improve immune cell recognition and enhance ADCC (an important NK cell-mediated cell lysis mechanism). It has been demonstrated that the combination of IMiD with a monoclonal antibody having Fc-like functions similar to CLN-619 has a synergistic effect in enhancing NK cell activity in a clinical setting. Furthermore, higher levels of sMICA were associated with decreased overall survival in MM patients compared to those who were sMICA-negative. 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, thereby leading to improved anti-myeloma activity.

[0753] Therefore, based on their respective modes of action, this embodiment proposes that using CLN-619 alone or in combination with pomalidomide and dexamethasone to treat multiple myeloma can enhance the anti-myeloma effects of both innate and adaptive immunity. Preferably, the method envisioned in this embodiment provides an effective treatment for relapsed / refractory multiple myeloma (R / R MM).

[0754] This embodiment details an open-label, dose-exploration study aimed at addressing the safety, tolerability, and efficacy of CLN-619 monotherapy and its combination with pomalidomide and dexamethasone for the treatment of recurrent / recurrent MM.

[0755] Part 1A of the study will use two escalating dose designs, with a 3-week DLT observation period for CLN-619 monotherapy (cycle 1) and in Part 1C, a 3-week DLT observation period for CLN-619 combination therapy (cycle 4 or earlier). The optimal dose of CLN-619 will be determined using the implementation of the 3+3 cohort design in Part 1. CLN-619 monotherapy in Part 1A will be examined first, and preliminary efficacy will be assessed before patients move to dose extensions for monotherapy (Part 1B) or combination therapy (Part 1C). Clinical response data from the first 4 cycles or shorter of CLN-619 monotherapy in Part 1A will be used to determine patient participation in Part 1B or 1C. Patients with a very good partial response (VGPR) or better, according to the International Myeloma Working Group (IMWG) response criteria, will continue to receive the designated dose of CLN-619 monotherapy (Part 1B). According to the IMWG response criteria, patients with disease progression (PD), partial response (PR), minimum response (MR), or stable disease (SD) will continue to receive their designated dose of CLN-619 in combination with pomalidomide and dexamethasone (partial 1C).

[0756] Based on all the data from Part 1 of the SRC review, Part 2 will begin the combination group of CLN-619 with pomalidomide + dexamethasone starting from the first cycle.

[0757] Dosage basis for treating relapsed / relapsed multiple myeloma

[0758] Based on 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, this study selected a starting dose of CLN-619 of 3 mg / kg Q3W.

[0759] Studies of patients with locally advanced or metastatic solid tumors who were recruited into the monotherapy group of the CLN-619-001 study and had received at least one dose of CLN-619 monotherapy provided a basis for MM dosing. In the aforementioned study, the safety, efficacy, and pharmacokinetics (PK) of CLN-619 were investigated in patients with advanced solid tumors whose disease had progressed after existing therapy, across a dose range of 0.1 mg / kg to 10 mg / kg. No disease-to-lack (DLT) was observed in any patient. In this dose group, only one patient experienced a TRAE (grade 2 IRR at the first CLN-619 infusion), and no serious TRAEs occurred.

[0760] Early observations support the efficacy of CLN-619 as a monotherapy in advanced or metastatic solid tumors (i.e., the solid tumors described in Examples 2 and 3 above), and the results are encouraging. Confirmed objective responses (RECIST v1.1) were observed at dose levels ≥3 mg / kg: one CR in a patient with parotid gland cancer treated at a dose level of 3 mg / kg, and two PRs in patients with endometrial cancer treated at dose levels of 3 and 10 mg / kg. Stable disease lasting at least 6 cycles was also observed at dose levels of 1 mg / kg and above.

[0761] In the CLN-619-001 study, preliminary pharmacokinetics of CLN-619 and sMICA were also determined (i.e., those of Examples 2 and 3 above). At steady state (cycle 3), exposure increases from 0.1 mg / kg to 1 mg / kg were greater than dose-proportional, but increases from 1 mg / kg to 10 mg / kg were approximately dose-proportional. The mean half-life of CLN-619 across dose groups ranged from 60.5 to 464 hours (2.5 to 19 days), with longer half-lives generally observed at higher doses and associated with dose-related decreases in systemic clearance, likely a result of target-mediated clearance (TMDD) saturation.

[0762] A dose-related increase in fold changes in serum sMICA concentrations was observed following CLN-619 treatment. Individual profiles of sMICA generally showed a steady increase in serum concentrations with continued CLN-619 treatment, particularly at higher dose levels. CLN-619 binding to sMICA leads to an increase in total sMICA serum concentrations, which may result in an overall reduction in free sMICA. Given the immunosuppressive effects of sMICA in the tumor microenvironment, reducing free sMICA concentrations may contribute to the clinical activity of CLN-619. TMDD saturation and sMICA pharmacokinetic responses indicate that near-complete target saturation can be achieved at dose levels starting from 3 mg / kg.

[0763] In solid tumors and multiple myeloma, MICA / MICB expression is typically elevated. Serum sMICA concentrations also tend to be elevated in patients with solid tumors and multiple myeloma, and higher concentrations are associated with poor prognosis. Some studies have reported higher serum sMICA concentrations in multiple myeloma patients than in solid tumor patients, which may reflect increased dropout rates and / or a higher overall tumor burden. Therefore, a dose of at least 3 mg / kg is recommended to achieve target saturation in MM patients.

[0764] In summary, CLN-619 was well tolerated in patients with solid tumors at doses up to 10 mg / kg, with objective responses observed only at doses of 3 mg / kg or higher. Evidence of target saturation (saturation of TMDD and pharmacodynamics of total sMICA in serum) appears to occur at approximately 3 mg / kg or higher in most patients. Therefore, in patients with multiple myeloma, a starting dose of 3 mg / kg for CLN-619 according to a Q3W dosing schedule is expected to be a safe and effective approach to targeted engagement. For overall study design, see [link to study design]. Figure 31 .

[0765] Combination therapy of CLN-619 and IMiD for R / R multiple myeloma .

[0766] Pomalidomide, marketed in the United States as Pomalyst®, is a thalidomide analog or immunomodulatory (IMiD) agent manufactured by Bristol Myers Squibb for the treatment of multiple myeloma (MM). When taken in combination with dexamethasone, pomalidomide exerts its effects by inducing tumor cell apoptosis, enhancing T-cell and NK-cell-mediated immunity, and inhibiting pro-inflammatory cytokines.

[0767] This study used a standard nursing dose of 4 mg and will allow for dose reduction to minimize toxicity observations of neutropenia and pneumonia observed in other studies.

[0768] Figure 31 The overall study design for treating recurrent / recurrent myocardial infarction (R / R MM) is shown. Because CLN-619 follows a 21-day dosing schedule, the inventors designed a shorter pomalidomide dosing schedule to match that of CLN-619. The standard care dosing regimen for pomalidomide is typically 28 days, with 21 days of dosing followed by a one-week break. The current study design will administer pomalidomide on days 1-14 and discontinue it on days 15-21 within each 21-day treatment cycle.

[0769] The patient will be treated with a combination of pomalidomide, dexamethasone, and CLN-619 until disease progression, intolerable toxicity, or up to 2 years (whichever occurs first).

[0770] Dexamethasone is a corticosteroid used to treat a variety of diseases by suppressing the immune system. Dexamethasone is often used in combination with other drugs to treat multiple myeloma.

[0771] Different doses of dexamethasone have been used in combination with pomalidomide, such as weekly doses of 20 mg (≥75 years) or 40 mg (<75 years), and have shown efficacy in the treatment of R / R MM.

[0772] In combination with pomalidomide and CLN-619, the current study design will administer dexamethasone at 20 mg PO or IV on days 1, 8, and 15 of each 21-day treatment cycle, regardless of age.

[0773] The patient will be treated with dexamethasone in combination with CLN-619 and pomalidomide until disease progression, intolerable toxicity, or up to 2 years (whichever occurs first).

[0774] During the treatment cycle, patients who meet all eligibility criteria will undergo a pre-dose assessment upon their Day 1 visit. Following the completion of the Day 1 pre-dose assessment, the first dose of study treatment (CLN-619) will be administered under field observation, followed by a post-dose assessment. Depending on the section and group, the study treatment will consist of an intravenous infusion of CLN-619.

[0775] In Part 1A (dose escalation), treatment will consist of the first four cycles of CLN-619 monotherapy. Patients with R / RMM will be treated with CLN-619 monotherapy at escalating doses using a standard 3+3 dose escalation design. CLN-619 will be administered at 3 and 6 mg / kg according to a 21-day cycle. Prophylactic medication will be provided on day 1 of the first treatment cycle and subsequent treatment cycles to reduce infusion-related reactions.

[0776] The decision-making rules guiding dose escalation decisions and study implementation will be based on DLT or MAD occurring in patients during the first three weeks of CLN-619 monotherapy. Any DLT occurring in subsequent cycles will be considered in the recommended optimal dose decision.

[0777] A 3+3 design will be used to evaluate toxicity in each cohort. After completing the 3+3 cohorts (minimum three, maximum six patients), if:

[0778] • None of the three patients experienced DLT; dose escalation can proceed to the next cohort.

[0779] • One of the three patients has a history of DLT; the other three patients will be recruited to this group.

[0780] • ≥2 patients have experienced DLT; exceeding MTD, and further recruitment in this group will be stopped.

[0781] ○ Then, the lower dose group, including intermediate doses, can be further explored until the MTD is determined.

[0782] Patients who are unable to complete the DLT observation period due to reasons other than experiencing DLT will be excluded when estimating parameters; however, all patient data will be included in the safety summary and data list.

[0783] Part 1B and 1C CLN-619 Dosage Extension: Patients who have completed the first four cycles of CLN-619 monotherapy in Part 1A and demonstrated acceptable safety and tolerability will proceed to Part 1B or 1C based on their efficacy assessment results.

[0784] Part 1B CLN-619 monotherapy: Patients with VGPR or better after four treatment cycles, based on IMWG response criteria, will continue with their specific monotherapy dose of CLN-619 in the fifth cycle and beyond.

[0785] Partial 1C CLN-619 + Pomalidomide + Dexamethasone: Patients with PR, MR, or SD according to IMWG response criteria after four cycles of CLN-619 monotherapy will begin CLN-619 in combination with pomalidomide and dexamethasone starting from cycle 5. Patients with PD according to IMWG criteria at any time after completing cycles 1–4 will receive CLN-619 in combination with pomalidomide and dexamethasone. The DLT observation period will occur during the first cycle of combination therapy (days 1–21). CLN-619 will be administered according to a 21-day cycle. Pomalidomide will be administered orally at 4 mg daily starting from day 1 of cycle 4, repeating the 21-day cycle from day 1 to day 14. Dexamethasone will be administered orally or intravenously at 20 mg on days 1, 8, and 15 of each cycle. Partial 1C patients will be closely monitored for DLT during the first three weeks of combination therapy.

[0786] At any time during a partial 1B or 1C study, if >30% of patients experience adverse events (AEs) that meet the DLT criteria during and after the defined DLT observation period, or if safety findings suggest clinically significant cumulative toxicity, the SRC may be convened to review data from the extended cohort and make recommendations regarding continued dosing for patients.

[0787] Part 2: Following a review of the available safety and efficacy data from Part 1, a single-dose combination of CLN-619 with pomalidomide and dexamethasone will be selected to enroll patients in Part 2, with the combination of CLN-619 and pomalidomide / dexamethasone to be administered from the first treatment cycle. CLN-619 will be administered according to a 21-day cycle. Pomalidomide will be administered orally at a dose of 4 mg daily from day 1 to day 14 of repeated 21-day cycles. Dexamethasone will be administered orally or intravenously at a dose of 20 mg on days 1, 8, and 15 of each cycle. At any time during the Part 2 study period, if >30% of patients experience adverse events (AEs) meeting the DLT criteria, or if safety findings suggest clinically significant cumulative toxicity, the SRC may be convened to review data from the extended cohort and make recommendations regarding continued dosing for patients.

[0788] References

[0789]

[0790]

[0791]

[0792]

[0793] All features disclosed in the specification (including claims, abstract, drawings, and appendices) and all steps in any disclosed method or process may be combined in any combination, except that at least some of such features and / or steps are mutually exclusive combinations. Each feature disclosed in the specification (including claims, abstract, drawings, and appendices) may be replaced by an alternative feature serving the same, equivalent, or similar purpose, unless expressly stated otherwise. Therefore, unless expressly stated otherwise, each disclosed feature is merely one example of a series of equivalent or similar features.

[0794] While certain preferred embodiments of the invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Various modifications, variations, and substitutions will occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments described herein may be employed. This means that the following claims define the scope of the invention and the methods and structures within the scope of these claims and their equivalents are therefore covered.

Claims

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

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

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

4. The method of claim 1, wherein the subject has not previously been treated with a PD-1 inhibitor (e.g., an anti-PD-1 antibody).

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

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

7. A method for treating parotid gland cancer 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).

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

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

10. The method of any of the preceding claims, wherein the cancer is mutated for one or more of the following: EGFR, TP53, HER2, PTEN, ARID1A, CTNNB1, or PLK3R1.

11. A method of treating a subject with cancer, the method comprising administering to the subject a therapeutically effective amount of an antibody binding to MICA / B (anti-MICA / B antibody), wherein the subject has previously been treated with: (i) VEGF inhibitors; (ii) EGFR inhibitors; and / or (iii) Hormone therapy.

12. The method of claim 11, wherein the subject responds to the VEGF inhibitor, the EGFR inhibitor, or the hormone therapy (e.g., a complete or partial response), wherein 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 of claim 12, wherein the subject's disease progresses after the response period to the VEGF inhibitor, the EGFR inhibitor, or the hormone therapy.

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

15. The method of claim 11, wherein the subject has stable disease after administration of the VEGF inhibitor, the EGFR inhibitor, or the hormone therapy, wherein optionally the subject has a stable disease 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 of claim 11, wherein treatment with the VEGF inhibitor, the EGFR inhibitor, or the hormone therapy is discontinued before the administration of the anti-MICA / B antibody is initiated.

17. The method of claim 11, wherein treatment with the VEGF inhibitor, the EGFR inhibitor, or the hormone therapy continues after the initiation of administration of the anti-MICA / B antibody.

18. The method of claim 11, wherein the subject's disease progresses after treatment with the VEGF inhibitor, the EGFR inhibitor, or the hormone therapy.

19. The method of any of the preceding claims, wherein after administration of the anti-MICA / B antibody, the subject has a response (e.g., a complete or partial response) or has a stable disease.

20. The method of claim 19, wherein the response following 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.

21. The method of any one of claims 11-20, 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 neximab).

22. The method of 11-20, 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 of 11-20, 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 antiadrenergic drug, or an estrogen receptor antagonist.

24. A method of treating a subject with cancer, the method comprising administering to the subject a therapeutically effective amount of an antibody binding 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), wherein: (i) The subject responded to the PD-1 inhibitor (e.g., with a complete or partial response); or (ii) The subject had stable disease after administration of the PD-1 inhibitor.

25. The method of 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 of claim 25, wherein the subject's disease progresses after the response period to the PD-1 inhibitor.

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

28. The method of claim 24, wherein, The subjects mentioned above had a stable disease 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 of claim 24, wherein treatment with the PD-1 inhibitor is discontinued before the administration of the anti-MICA / B antibody is initiated.

30. The method of claim 24, wherein treatment with the PD-1 inhibitor continues after the administration of the anti-MICA / B antibody is initiated.

31. The method as described in any of the preceding claims, wherein after administration of the anti-MICA / B antibody, the subject has a response (e.g., a complete or partial response) or has a stable disease.

32. The method of claim 37, wherein the response following 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.

33. The method of any of the preceding claims, wherein the PD-1 inhibitor is an anti-PD-1 antibody.

34. The method of any of the preceding claims, wherein the anti-PD-1 inhibitor comprises pimiralimumab, pembrolizumab, nivolumab, cimipril, AMP-224, APM-514 or spartazolizumab.

35. The method of any of the preceding 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, the method comprising: (i) Administering a first dose of the MICA / B-binding antibody (anti-MICA / B antibody) to the subject, and (ii) Administer a therapeutically effective amount of corticosteroid to the subject prior to the first dose of the anti-MICA / B antibody.

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

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

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

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

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

42. The method of 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 of any one of claims 36-42, wherein the corticosteroid is dexamethasone.

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

45. The method of claim 44, wherein: The female reproductive tract tumors include endometrial tumors, ovarian cancer, or cervical cancer; The salivary gland tumors include mucoepidermoid tumors such as parotid gland tumors.

46. ​​The method of any of the preceding claims, wherein the subject has cancer, said cancer being parotid gland cancer, cervical cancer, endometrial cancer, breast cancer, colon cancer, ovarian cancer, prostate cancer, sarcoma, melanoma, adenoid cystic salivary gland tumor, peritoneal mesothelioma, rectal squamous cell carcinoma, leiomyosarcoma, colorectal cancer, kidney cancer, thyroid cancer, NSCLC, duodenal cancer, pancreatic cancer, mediastinal endometrial sarcoma, head and neck cancer, or cecal cancer.

47. The method of any of the preceding claims, wherein the subject suffers from cancer characterized by one or more of the following: Cancers characterized by low levels of PD-L1 expression; Cancers characterized by high levels of PD-L1 expression; Cancers characterized by low tumor mutational burden (TMB); Cancers characterized by high tumor mutational burden (TMB); Immune cold cancer; Immune fever cancer; Hormone-sensitive cancers; Cancers characterized by the overexpression of carcinogenic drivers; or Cancers expressing one or more of the following: EGFR, ER, PR, or HER2.

48. The method of any of the preceding claims, wherein the subject has metastatic cancer.

49. The method of any of the preceding claims, wherein the subject has previously received a therapy including chemotherapy.

50. The method of any of the preceding claims, wherein the anti-MICA / B antibody is administered at a dose of about 3 mg / kg to about 10 mg / kg.

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

52. The method of any of the preceding claims, wherein the dosing interval comprises a three-week cycle, and wherein the anti-MICA / B antibody is administered once every three weeks (Q3W).

53. The method of claim 51 or 52, wherein 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.

54. The method of claim 53, wherein the repeated dosing interval is 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.

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

6.

56. The method of claim 55, wherein the 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 with it.

57. The method of claim 55 or 56, wherein the 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 with it.

58. The method of any one of claims 55-57, wherein 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 with it.

59. The method of any one of claims 55-58, wherein 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 with it.

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

61. The method of any of the preceding claims, wherein the anti-MICA / B antibody specifically binds to the α-3 domain of the MICA protein, the MICB protein, or both the MICA and MICB proteins.

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

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

64. The method of any of the preceding claims, wherein the anti-MICA / B antibody is selected from intact immunoglobulins, scFv, Fab, F(ab')2 or disulfide-linked Fv.

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

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

67. The method as claimed in any of the preceding claims, wherein the method further comprises administering an effective amount of a PD-1 inhibitor to the subject.

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

69. The method of claim 67 or 68, wherein the anti-PD-1 inhibitor comprises pimiralimumab, pembrolizumab, nivolumab, cimiprimab, AMP-224, APM-514, or spartazolizumab.

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

71. The method of claim 70, further comprising administering a therapeutically effective amount of the immunomodulatory drug to the subject.

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

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

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

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

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

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

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

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

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

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

82. The method of 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 of any one of claims 76-82, wherein the corticosteroid is dexamethasone.

Images