Cancer Treatment

Abstract

Use of LAG-3 protein or derivatives thereof for the treatment of cancer in a subject is described. In certain embodiments, the subject has one or more of low monocyte count, luminal B breast cancer, age less than about 85 years, previously treated with a CDK4 / 6 inhibitor, not previously treated with taxane chemotherapy, elevated neutrophil to lymphocyte ratio, and diagnosed less than about 5 years ago. In another embodiment, the LAG-3 protein or derivative is administered to the subject at a dosage of >30 mg to <120 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321. In a further embodiment, the LAG-3 protein or derivative is administered to the subject on the same day as the chemotherapeutic agent. In another embodiment, the LAG-3 protein or derivative is administered to the subject one or more times in the absence of the chemotherapeutic agent after one or more doses of the LAG-3 protein or derivative have been administered to the subject before, with, or after one or more doses of the chemotherapeutic agent. In a further aspect, the subject is a hormone receptor positive HER2-negative / low (HR + Patients with HER2-negative / low) metastatic breast cancer or metastatic triple-negative breast cancer (TNBC).

Description

[Technical field]

[0001] The present invention relates to the use of LAG-3 protein or a derivative thereof for the treatment of cancer. [Background technology]

[0002] Over the past decade, PD-1 and CTLA-4 immune checkpoint inhibitors such as OPDIVO (nivolumab), KEYTRUDA (pembrolizumab) and YERVOY (ipilimumab) have become standard of care for many forms of cancer, but unfortunately, many patients still do not respond to these medications. In an effort to improve patient outcomes, significant research has been conducted to investigate other immune checkpoints such as LAG-3, TIM-3, VISTA, CD47, IDO and TIGIT. LAG-3 in particular has emerged as a promising checkpoint, and many companies are developing new inhibitors targeting this checkpoint. The goal of LAG-3 inhibitors, like the currently approved PD-1 and CTLA-4 inhibitors, is to block downregulation of the immune system, i.e., to "take the brakes off" the body's immune processes. Significant research is also being conducted to explore combinations of PD-1 and CTLA-4 immune checkpoint inhibitors with other approved or experimental therapies. Another type of active immunotherapy being investigated is antigen-presenting cell (APC) activators. APC activators bind to antigen-presenting cells, such as dendritic cells, monocytes and macrophages, via MHC II molecules. This activates the APC to become professional antigen-presenting cells, thereby presenting antigens to the adaptive immune system. This leads to the activation and proliferation of CD4+ (helper) and CD8+ (cytotoxic) T cells. Thus, the purpose of APC activators is to "step on the gas" on the body's immune system.

[0003] Eftilagimod alpha (IMP 321 or efti), a soluble dimeric recombinant form of LAG-3, is a first-in-class APC activator in clinical development. By stimulating dendritic cells and other APCs via MHC class II molecules, IMP321 induces potent anti-cancer T cell responses. IMP321 is described in WO 2009 / 044273, which also describes the use of IMP321 alone and in combination with chemotherapeutic agents for the treatment of cancer. There remains a need in the art for improved cancer treatments and treatment regimens that result in better patient outcomes. This is particularly true for cancers where patients treated with current pharmaceutical agents have a poor prognosis.

[0004] Metastatic breast cancer (MBC) represents a major treatment challenge, either as a de novo breast cancer diagnosis or as a recurrence after previous treatment for early stage disease. The goal of MBC treatment is primarily disease control and palliation of symptoms with minimal side effects.

[0005] Endocrine therapy (ET), in combination with CDK4 / 6 inhibitors, other targeted therapies, or alone, has been shown to be effective in treating hormone receptor-positive (HR) + ), remains the mainstay in the management of HER2-negative / low MBC. However, the likelihood of developing ET resistance is high. Before the introduction of CDK4 / 6 inhibitors into the treatment landscape, patients with ET resistance were usually prescribed single-agent chemotherapy with a median overall survival (OS) of approximately 24 months. The addition of CDK4 / 6 inhibitors to ET-based treatment reduced the OS of subsequent chemotherapy, with the median OS decreasing to approximately 12-18 months. HR, which is resistant to endocrine-based therapy and eligible for chemotherapy, + There is high unmet medical need in HER2-negative / low MBC patients.

[0006] Metastatic triple-negative breast cancer (TNBC) is an aggressive disease with poor outcome. This type of breast cancer is characterized by the absence of estrogen receptor (ER), progesterone receptor (PR) and HER2 expression, as well as the presence of high histological grade and mitotic rate. The choice of therapy depends mainly on PD-L1 expression. For patients with PD-L1-positive tumors (approximately 40% of all TNBC patients), anti-PD-1 + chemotherapy is recommended. Otherwise, chemotherapy remains the main systemic treatment, and international guidelines support the use of single-agent taxanes (with or without bevacizumab) or anthracyclines as first-line therapy, with a median OS of up to 18 months. Given the suboptimal results with chemotherapy alone (median OS between 15 and 18 months) or immune checkpoint inhibitors (ICI; 21 to 25 months) for PD-L1-positive tumors, there is a high unmet medical need for these patients. Summary of the Invention

[0007] In one embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for use in the prevention, treatment or amelioration of cancer in a subject with low monocyte counts.

[0008] In another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject with low monocyte count.

[0009] In yet another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for the prevention, treatment, or amelioration of cancer in a subject with low monocyte count.

[0010] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject with low monocyte count, comprising the step of administering LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, to a subject in need of such prevention, treatment or amelioration.

[0011] In one embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for use in the prevention, treatment, or amelioration of luminal B breast cancer in a subject.

[0012] In another embodiment, the invention relates to the use of a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment, or amelioration of luminal B breast cancer in a subject.

[0013] In yet another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for the prevention, treatment, or amelioration of luminal B breast cancer in a subject.

[0014] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating luminal B breast cancer, comprising administering LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, to a subject in need of such prevention, treatment or amelioration.

[0015] In one embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for use in the prevention, treatment, or amelioration of cancer in a subject less than about 85 years of age.

[0016] In another embodiment, the present invention relates to the use of a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject less than about 85 years of age.

[0017] In yet another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for the prevention, treatment, or amelioration of cancer in a subject less than about 85 years of age.

[0018] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject less than about 85 years of age, comprising administering to a subject in need of such prevention, treatment or amelioration a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule.

[0019] In one embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for use in the prevention, treatment, or amelioration of cancer in a subject previously treated with a CDK4 / 6 inhibitor.

[0020] In another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject previously treated with a CDK4 / 6 inhibitor.

[0021] In another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for the prevention, treatment, or amelioration of cancer in a subject previously treated with a CDK4 / 6 inhibitor.

[0022] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject previously treated with a CDK4 / 6 inhibitor, comprising administering a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, to a subject in need of such prevention, treatment or amelioration.

[0023] In one embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for use in the prevention, treatment, or amelioration of cancer in a subject not previously treated with taxane chemotherapy.

[0024] In another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment or amelioration of cancer in a subject who has not previously been treated with taxane chemotherapy.

[0025] In yet another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for the prevention, treatment, or amelioration of cancer in a subject who has not previously been treated with taxane chemotherapy.

[0026] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject who has not previously been treated with taxane chemotherapy, comprising administering LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, to a subject in need of such prevention, treatment or amelioration.

[0027] In one embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for use in the prevention, treatment, or amelioration of cancer in a subject with an elevated neutrophil to lymphocyte ratio.

[0028] In another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject with an elevated neutrophil to lymphocyte ratio.

[0029] In yet another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to MHC class II molecules, for the prevention, treatment, or amelioration of cancer in a subject with an elevated neutrophil to lymphocyte ratio.

[0030] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject having an elevated neutrophil to lymphocyte ratio, comprising administering to a subject in need of such prevention, treatment or amelioration a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule.

[0031] In one embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for use in the prevention, treatment, or amelioration of cancer in a subject diagnosed less than about 5 years ago.

[0032] In another embodiment, the present invention relates to the use of a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject diagnosed less than about 5 years ago.

[0033] In yet another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for the prevention, treatment, or amelioration of cancer in a subject diagnosed less than about 5 years ago.

[0034] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject diagnosed less than about 5 years ago, comprising administering a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, to a subject in need of such prevention, treatment or amelioration.

[0035] In a further embodiment, the present invention provides a LAG-3 protein or a derivative thereof capable of binding to an MHC class II molecule for use in preventing, treating or ameliorating cancer in a subject, wherein the LAG-3 protein or derivative thereof is administered to a subject at a dosage of >30 mg to <200 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0036] In a further embodiment, the present invention provides a LAG-3 protein or a derivative thereof capable of binding to an MHC class II molecule for use in preventing, treating or ameliorating cancer in a subject, wherein the LAG-3 protein or derivative thereof is administered to a subject at a dosage of >30 mg to <120 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0037] In a further embodiment, the present invention provides the use of a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment or amelioration of cancer in a subject, wherein the LAG-3 protein or derivative is administered to the subject at a dosage of >30 mg to <200 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0038] In a further embodiment, the present invention provides the use of a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment or amelioration of cancer in a subject, wherein the LAG-3 protein or derivative is administered to the subject at a dosage of >30 mg to <120 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0039] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject, comprising administering to the subject a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, at a dosage of >30 mg to <200 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0040] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject, comprising administering to the subject a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, at a dosage of >30 mg to <120 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0041] In a further embodiment, the present invention provides a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for use in preventing, treating or ameliorating cancer in a subject, wherein the LAG-3 protein or derivative thereof is administered to the subject on the same day as a chemotherapeutic agent.

[0042] In a further embodiment, the present invention provides the use of a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment or amelioration of cancer in a subject, wherein the LAG-3 protein or derivative is administered to the subject on the same day as a chemotherapeutic agent.

[0043] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject, comprising administering to the subject a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, on the same day as a chemotherapy agent.

[0044] Optionally, the LAG-3 protein or derivative is administered to the subject on days 1 and 15 of a 4-week cycle, and the chemotherapeutic agent is administered to the subject on days 1, 8, and 15 of the 4-week cycle, and optionally, the 4-week cycle is repeated for 4 to 8 cycles, preferably 6 cycles.

[0045] In a further embodiment, the present invention provides a LAG-3 protein or a derivative thereof capable of binding to an MHC class II molecule for use in the prevention, treatment, or amelioration of cancer in a subject, wherein the LAG-3 protein or derivative is administered to the subject one or more times in the absence of a chemotherapeutic agent after one or more doses of the LAG-3 protein or derivative have been administered to the subject before, with, or after one or more doses of a chemotherapeutic agent.

[0046] In a further embodiment, the present invention provides the use of a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment or amelioration of cancer in a subject, wherein the LAG-3 protein or derivative is administered to the subject one or more times in the absence of a chemotherapeutic agent after one or more doses of the LAG-3 protein or derivative have been administered to the subject prior to, along with or after one or more doses of a chemotherapeutic agent.

[0047] In a further embodiment, the present invention provides a method for preventing, treating, or ameliorating cancer in a subject, comprising administering to the subject one or more doses of LAG-3 protein or derivative capable of binding to an MHC class II molecule in the absence of a chemotherapeutic agent, after which one or more doses of LAG-3 protein or derivative have been administered to the subject prior to, with, or after one or more doses of a chemotherapeutic agent.

[0048] In a further embodiment, the present invention relates to a LAG-3 protein or a derivative thereof capable of binding to an MHC class II molecule for use in the prevention, treatment or amelioration of metastatic breast cancer in a subject, the subject being hormone receptor positive, HER2-negative / low (HR + The present invention provides a LAG-3 protein or derivative thereof for use in patients with HER2-negative / low (HER2-negative / low) metastatic breast cancer, or metastatic triple-negative breast cancer (TNBC) patients.

[0049] In a further embodiment, the present invention relates to the use of a LAG-3 protein or a derivative thereof capable of binding to an MHC class II molecule in the manufacture of a medicament for the prevention, treatment or amelioration of metastatic breast cancer in a subject, the subject being hormone receptor positive, HER2-negative / low (HR + The present invention provides for use in patients with HER2-negative / low (HER2-negative / low) metastatic breast cancer or metastatic triple-negative breast cancer (TNBC).

[0050] In a further embodiment, the present invention provides a method of preventing, treating, or ameliorating metastatic breast cancer in a subject, comprising administering to the subject a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, wherein the subject is a hormone receptor positive, HER2-negative / low (HR + The present invention provides a method for treating a patient with HER2-negative / low (HER2-negative / low) metastatic breast cancer or a patient with metastatic triple-negative breast cancer (TNBC). [Brief description of the drawings]

[0051] [Figure 1] The amino acid sequence of the mature human LAG-3 protein (SEQ ID NO:1) is shown. The four extracellular Ig superfamily domains are amino acid residues: 1-149 (D1); 150-239 (D2), 240-330 (D3), and 331-412 (D4). The amino acid sequence of the extra loop structure of the D1 domain of the human LAG-3 protein is shown in bold underlined type (SEQ ID NO:2). [Diagram 2]Major breast cancer subtypes by HR and HER2 status (Source: Administration,FaD2022.FDA Approves First Targeted Therapy for HER2-Low Breast Cancer FDA.pdf). [Diagram 3] HR+HER2-negative / low MBC treatment algorithm shown (Source: Adapted from Borges, VF2021. Options for Endocrine-Refractory, Hormone Receptor-Positive Breast Cancer: Which Target and When? American Society of Clinical Oncology). [Figure 4] A summarized algorithm for the treatment of mTNBC based on the ESMO guidelines. [Diagram 5] The AIPAC-003 study flowchart is shown. [Figure 6] An algorithm for defining the optimal biological dose is presented. [Figure 7] Kaplan-Meier curves for overall survival probability for patients with BOR of CR / PR (red) and BOR of PD+SD-NA (blue). BOR was assessed by RECIST 1.1. Exploratory analysis from AIPAC. [Figure 8] CD8+ T cell counts before and after treatment (left) and correlation with overall survival (right); AIPAC trial (Source: et al. 2021. Final results from AIPAC: A phase IIb trial comparing eftilagimod alpha(soluble LAG-3 protein) in combination with weekly paclitaxel in HR+HER2-MBC. Poster presented at SITC 2021). [Figure 9] Change in absolute lymphocyte count (ALC) from baseline (left) correlates with overall survival (right); and [Figure 10] A schematic diagram of the AIPAC-003 study is shown. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0052] Treatment strategies for patient subgroups with low initial monocyte counts In one embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for use in the prevention, treatment or amelioration of cancer in a subject with low monocyte counts.

[0053] In another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject with low monocyte count.

[0054] In yet another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for the prevention, treatment, or amelioration of cancer in a subject with low monocyte count.

[0055] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject with low monocyte count, comprising administering to a subject in need of such prevention, treatment or amelioration LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule.

[0056] Exemplary cancers that may be treated according to the present invention include, but are not limited to, breast cancer, skin cancer, lung cancer (especially NSCLC), ovarian cancer, renal cancer, colon cancer, colorectal cancer, gastric cancer, esophageal cancer, pancreatic cancer, bladder cancer, urothelial cancer, liver cancer, melanoma (e.g., metastatic malignant melanoma), prostate cancer (e.g., hormone refractory prostate cancer), head and neck cancer (e.g., head and neck squamous cell carcinoma), cervical cancer, thyroid cancer, glioblastoma, glioma, leukemia, lymphoma (e.g., B cell lymphoma), adrenal cancer, AIDS-related cancer, alveolar soft part sarcoma, astrocytoma, bone cancer, brain and spinal cord tumors, metastatic brain tumors, carotid ball tumors, chondrosarcoma, chordoma, chromophobe renal cell carcinoma, clear cell carcinoma, cutaneous benign fibrous histiocytoma, desmoplastic small cell tumor, Ependymoma, Ewing's sarcoma, extraskeletal myxoid chondrosarcoma, fibrous imperfecta osseous, fibrous dysplasia of bone, gallbladder or bile duct cancer, gestational trophoblastic disease, germ cell tumors, hematologic malignancies, hepatocellular carcinoma, islet cell tumors, Kaposi's sarcoma, renal cancer, lipoma / benign lipomatous tumors, liposarcoma / malignant lipomatous tumors, medulloblastoma, meningioma, Merkel cell tumor, multiple endocrine neoplasia, multiple myeloma, myelodysplastic syndromes, neuroblastoma, neuroendocrine tumors, papillary thyroid cancer, parathyroid tumors, childhood cancer, peripheral nerve sheath tumors, pheochromocytoma, pituitary tumors, prostate cancer, posterior uveal melanoma, rare hematologic disorders, renal metastatic cancer, rhabdoid tumor, rhabdomyosarcoma, sarcoma, soft tissue sarcoma, squamous cell carcinoma, gastric cancer, synovial sarcoma, testicular cancer, thymic carcinoma, thymoma, metastatic cancer of the thyroid, and uterine cancer.

[0057] In one embodiment, the cancer is breast cancer. Suitably, the breast cancer is an adenocarcinoma of the breast.

[0058] According to embodiments of the invention, the cancer may have progressed to metastatic disease.

[0059] In another embodiment, the breast cancer is a hormone receptor positive cancer (estrogen receptor positive and / or progesterone receptor positive), which can be HER2 positive or HER2 negative. In one embodiment, the hormone receptor positive cancer is HER2 negative. The hormone receptor positive cancer can be hormone receptor positive metastatic breast cancer. In one embodiment, the hormone receptor positive cancer is hormone receptor positive HER2 negative metastatic breast cancer.

[0060] In one embodiment, the hormone receptor positive cancer is luminal B breast cancer.Luminal B breast cancer is hormone receptor positive, either HER2 positive or HER2 negative, and has high levels of Ki-67.Luminal B cancer generally grows slightly faster than luminal A cancer, and has a slightly worse prognosis.

[0061] In one embodiment, hormone receptor positive cancer is HER2 negative and luminal B subtype.As in other embodiments of the present invention, hormone receptor positive HER2 negative breast cancer with luminal B subtype may progress to metastatic disease.Therefore, in one embodiment, hormone receptor positive cancer is hormone receptor positive HER2 negative metastatic breast cancer with luminal B subtype.

[0062] In another embodiment, the hormone receptor positive cancer is luminal A breast cancer. Luminal A breast cancer is hormone receptor positive (estrogen receptor positive and / or progesterone receptor positive), HER2 negative, and has low levels of the protein Ki-67.

[0063] In yet another embodiment, the breast cancer is triple-negative breast cancer (estrogen receptor negative, progesterone receptor negative and HER2 negative).

[0064] In one particular embodiment, the breast cancer is hormone receptor positive, HER2 negative / low (HR + / HER2-negative / low) breast cancer or triple-negative breast cancer (TNBC).

[0065] Preferably, the breast cancer is hormone receptor positive, HER2 negative / low (HR + / HER2-negative / low) metastatic breast cancer or metastatic triple-negative breast cancer (TNBC).

[0066] In a further embodiment, the breast cancer is HER2-enriched breast cancer.HER2-enriched breast cancer is hormone receptor negative (estrogen receptor negative and progesterone receptor negative) and HER2 positive.HER2-enriched breast cancer tends to grow faster than luminal cancer and may have a worse prognosis.

[0067] In some embodiments, the LAG-3 protein or derivative thereof is administered parenterally, including by subcutaneous, intravenous or intramuscular injection. In certain embodiments, the LAG-3 protein or derivative thereof is administered subcutaneously by injection.

[0068] According to certain embodiments of the invention, patients with low initial monocyte counts are selected for treatment. As defined herein, "low monocyte count" refers to a baseline monocyte count of about 0.25×10 monocytes per L of blood. 9 "Baseline" means prior to initiation of treatment according to the present invention.

[0069] <Treatment methods for luminal B breast cancer subgroup> In one embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for use in the prevention, treatment, or amelioration of luminal B breast cancer in a subject.

[0070] In another embodiment, the invention relates to the use of a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment, or amelioration of luminal B breast cancer in a subject.

[0071] In yet another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for the prevention, treatment, or amelioration of luminal B breast cancer in a subject.

[0072] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating luminal B breast cancer in a subject, comprising administering to a subject in need of such prevention, treatment or amelioration a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule.

[0073] As described herein, luminal B breast cancers are hormone receptor positive, either HER2 positive or HER2 negative, and have high levels of Ki-67. Luminal B cancers generally grow slightly faster than luminal A cancers, and have a slightly worse prognosis.

[0074] In one embodiment, the luminal B breast cancer is HER2 negative. In another embodiment, the luminal B breast cancer is HER2 negative and has progressed to metastatic disease. Thus, in an embodiment of the present invention, the luminal B breast cancer is a HER2 negative metastatic breast cancer.

[0075] In one particular embodiment, the luminal B breast cancer is a hormone receptor positive HER2-negative / low metastatic breast cancer. Preferably, the luminal B breast cancer is a hormone receptor positive HER2-negative / low metastatic breast cancer.

[0076] In another embodiment, the luminal B breast cancer is HER2 positive. In another embodiment, the luminal B breast cancer is HER2 positive and has progressed to metastatic disease. Thus, in an embodiment of the invention, the luminal B breast cancer is a HER2 positive metastatic breast cancer.

[0077] In yet another embodiment, the subject has luminal B breast cancer and also has a low monocyte count.

[0078] In one embodiment, the subject has HER2 negative, luminal B breast cancer and also has a low monocyte count. In another embodiment, the subject has HER2 negative / low, luminal B breast cancer and also has a low monocyte count.

[0079] Alternatively, the subject has luminal B breast cancer that is HER2 positive and also has a low monocyte count.

[0080] In one particular embodiment, the subject has metastatic luminal B breast cancer that is HER2 negative and the subject also has a low monocyte count. In another embodiment, the subject has metastatic luminal B breast cancer that is HER2 negative / low and also has a low monocyte count.

[0081] Treatment strategies for age-based patient subgroups In one embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for use in the prevention, treatment, or amelioration of cancer in a subject less than about 85 years of age.

[0082] In another embodiment, the present invention relates to the use of a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject less than about 85 years of age.

[0083] In yet another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for the prevention, treatment, or amelioration of cancer in a subject less than about 85 years of age.

[0084] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject less than about 85 years of age, comprising administering to a subject in need of such prevention, treatment or amelioration a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule.

[0085] Suitably, the subject is less than about 85 years of age, less than about 80 years of age, less than about 75 years of age, less than about 70 years of age, less than about 65 years of age, less than about 60 years of age, less than about 55 years of age, less than about 50 years of age, less than about 45 years of age, or less than about 40 years of age.

[0086] In one embodiment, the subject is between about 18 and about 85 years of age. In another embodiment, the subject is between about 18 and about 80 years of age. In yet another embodiment, the subject is between about 18 and about 75 years of age. In a further embodiment, the subject is between about 18 and about 70 years of age. In yet a further embodiment, the subject is between about 18 and about 65 years of age. In one embodiment, the subject is between about 18 and about 60 years of age. In another embodiment, the subject is between about 18 and about 55 years of age. In yet another embodiment, the subject is between about 18 and about 50 years of age. In a further embodiment, the subject is between about 18 and about 45 years of age. In yet a further embodiment, the subject is between about 18 and about 40 years of age.

[0087] In another embodiment, the subject is premenopausal.

[0088] Suitably, the subject is less than about 85 years of age, less than about 84 years of age, less than about 83 years of age, less than about 82 years of age, less than about 81 years of age, less than about 80 years of age, less than about 79 years of age, less than about 78 years of age, less than about 77 years of age, less than about 76 years of age, less than about 75 years of age, less than about 74 years of age, less than about 73 years of age, less than about 72 years of age, less than about 71 years of age, less than about 70 years of age, less than about 69 years of age, less than about 68 years of age, less than about 67 years of age, less than about 66 years of age, less than about 65 years of age, less than about 64 years of age, less than about 63 years of age, The age is less than 62 years old, less than about 61 years old, less than about 60 years old, less than about 59 years old, less than about 58 years old, less than about 57 years old, less than about 56 years old, or less than about 55 years old, less than about 54 years old, less than about 53 years old, less than about 52 years old, less than about 51 years old, less than about 50 years old, less than about 49 years old, less than about 48 years old, less than about 47 years old, less than about 46 years old, less than about 45 years old, less than about 44 years old, less than about 43 years old, less than about 42 years old, less than about 41 years old, or less than about 40 years old.

[0089] In either case, the patient may optionally be greater than about 18 years of age and less than the ages described herein.

[0090] In one particular embodiment, the subject is less than about 65 years of age.

[0091] In another specific embodiment, the subject is less than about 53 years of age.

[0092] Exemplary cancers that may be treated according to this embodiment of the invention include, but are not limited to, those described hereinabove.

[0093] In one particular embodiment, the cancer is breast cancer. Suitably, the breast cancer is hormone receptor positive, HER2 negative / low (HR + / HER2-negative / low) breast cancer or triple-negative breast cancer (TNBC).

[0094] Preferably, the breast cancer is hormone receptor positive, HER2 negative / low (HR + / HER2-negative / low) metastatic breast cancer or metastatic triple-negative breast cancer (TNBC).

[0095] Treatment strategies in subgroups of patients previously treated with CDK4 / 6 inhibitors In one embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for use in the prevention, treatment, or amelioration of cancer in a subject previously treated with a CDK4 / 6 inhibitor.

[0096] In another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject previously treated with a CDK4 / 6 inhibitor.

[0097] In another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for the prevention, treatment, or amelioration of cancer in a subject previously treated with a CDK4 / 6 inhibitor.

[0098] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject previously treated with a CDK4 / 6 inhibitor, comprising administering a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, to a subject in need of such prevention, treatment or amelioration.

[0099] CDK4 / 6 inhibitors are a new class of treatments for cancer, particularly hormone receptor-positive, HER2-negative metastatic breast cancer, that target cyclin-dependent kinases 4 and 6. Exemplary CDK4 / 6 inhibitors include, but are not limited to, palbociclib, ribociclib, and abemaciclib.

[0100] Suitably, in one embodiment of the invention, the subject has previously been treated with a CDK4 / 6 inhibitor, but their disease continues to progress and requires an alternative treatment option.

[0101] Exemplary cancers that may be treated according to this embodiment of the invention include, but are not limited to, those described hereinabove.

[0102] In one embodiment, the cancer is breast cancer. In another embodiment, the breast cancer is hormone receptor positive HER2 negative (HR+ / HER2-) breast cancer or hormone receptor positive HER2-negative / low (HR + / HER2-negative / low) breast cancer.

[0103] Preferably, the breast cancer is hormone receptor positive, HER2 negative / low (HR + / HER2-negative / low) breast cancer.

[0104] In yet another embodiment, the hormone receptor positive, HER2 negative breast cancer is selected from the group consisting of hormone receptor positive, HER2 negative metastatic breast cancer and hormone receptor positive, HER2 negative / low (HR + / HER2-negative / low-grade) metastatic breast cancer.

[0105] Preferably, the breast cancer is hormone receptor positive, HER2 negative / low (HR + / HER2-negative / low-grade) metastatic breast cancer.

[0106] Treatment strategies in subgroups of patients not previously treated with taxane therapy In one embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for use in the prevention, treatment, or amelioration of cancer in a subject not previously treated with taxane chemotherapy.

[0107] In another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment or amelioration of cancer in a subject who has not previously been treated with taxane chemotherapy.

[0108] In yet another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for the prevention, treatment, or amelioration of cancer in a subject who has not previously been treated with taxane chemotherapy.

[0109] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject who has not previously been treated with taxane chemotherapy, comprising administering LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, to a subject in need of such prevention, treatment or amelioration.

[0110] Advantageously, in one embodiment of the present invention, the subject has not been previously treated with taxane chemotherapy.Taxane chemotherapy is characterized by taxadiene structure and acts by binding to tubulin, thereby stabilizing microtubule polymer and protecting it from degradation.This then blocks mitotic progression and induces apoptosis (cell death).Taxane chemotherapy is effective in a wide variety of cancers, including breast cancer, ovarian cancer, lung cancer, pancreatic cancer, prostate cancer, and head and neck cancer.

[0111] Exemplary taxane chemotherapies include, but are not limited to, paclitaxel, nab-paclitaxel, docetaxel, cabazitaxel, larotaxel, mirataxel, ortataxel, taxoplexin, opaxio, tesetaxel, and BMS-184476.

[0112] Exemplary cancers that may be treated according to this embodiment of the invention include, but are not limited to, those described hereinabove.

[0113] In one particular embodiment, the cancer is breast cancer. Suitably, the breast cancer is hormone receptor positive, HER2 negative / low (HR + / HER2-negative / low) breast cancer or triple-negative breast cancer (TNBC).

[0114] Preferably, the breast cancer is hormone receptor positive, HER2 negative / low (HR + / HER2-negative / low) metastatic breast cancer or metastatic triple-negative breast cancer (TNBC).

[0115] Treatment methods in patient subgroups with elevated neutrophil-to-lymphocyte ratios In one embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for use in the prevention, treatment, or amelioration of cancer in a subject with an elevated neutrophil to lymphocyte ratio.

[0116] In another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject with an elevated neutrophil to lymphocyte ratio.

[0117] In yet another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to MHC class II molecules, for the prevention, treatment, or amelioration of cancer in a subject with an elevated neutrophil to lymphocyte ratio.

[0118] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject having an elevated neutrophil to lymphocyte ratio, comprising administering to a subject in need of such prevention, treatment or amelioration a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule.

[0119] The neutrophil-to-lymphocyte ratio (NLR), calculated as a simple ratio between neutrophil and lymphocyte counts, is a biomarker that reflects the balance between two aspects of the immune system: acute and chronic inflammation (indicated by neutrophil count) and adaptive immunity (lymphocyte count). An elevated ratio reflects increased inflammation and has been associated with poor prognosis in patients with several types of cancer as well as other diseases, including cardiovascular, infectious, and inflammatory diseases. The normal NLR in healthy adult non-geriatric patients is between 0.78 and 3.53 (Forget P, Khalifa C, Defour JP, Latinne D, Van Pel MC, De Kock M. What is the normal value of the neutrophil-to-lymphocyte ratio? BMC Res Notes. 2017 Jan 3;10(1):12).

[0120] As defined herein, an "elevated neutrophil to lymphocyte ratio (NLR)" is a baseline greater than about 3.53. "Baseline" means prior to initiation of treatment according to the present invention.

[0121] In one embodiment, the subject's NLR at baseline is greater than about 3.53, greater than about 3.54, greater than about 3.55, greater than about 3.56, greater than about 3.57, greater than about 3.58, greater than about 3.59, greater than about 3.60, greater than about 3.61, greater than about 3.62, greater than about 3.63, greater than about 3.64, greater than about 3.65, greater than about 3.66, greater than about 3.67, greater than about 3.68, greater than about 3.69, or greater than about 3.70.

[0122] In one embodiment, the patient's baseline NLR is greater than about 3.65.

[0123] Exemplary cancers that may be treated according to this embodiment of the invention include, but are not limited to, those described hereinabove.

[0124] In one particular embodiment, the cancer is breast cancer. Suitably, the breast cancer is hormone receptor positive, HER2 negative / low (HR + / HER2-negative / low) breast cancer or triple-negative breast cancer (TNBC).

[0125] Preferably, the breast cancer is hormone receptor positive, HER2 negative / low (HR + / HER2-negative / low) metastatic breast cancer or metastatic triple-negative breast cancer (TNBC).

[0126] Treatment options for the subgroup of patients diagnosed less than 5 years ago In one embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for use in the prevention, treatment, or amelioration of cancer in a subject diagnosed less than about 5 years ago.

[0127] In another embodiment, the present invention relates to the use of a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject diagnosed less than about 5 years ago.

[0128] In yet another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for the prevention, treatment, or amelioration of cancer in a subject diagnosed less than about 5 years ago.

[0129] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject diagnosed less than about 5 years ago, comprising administering a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, to a subject in need of such prevention, treatment or amelioration.

[0130] In one embodiment, at the time of treatment, the subject was diagnosed with cancer less than about 5 years ago, less than about 4 years ago, less than about 3 years ago, less than about 2 years ago, or less than about 1 year ago.

[0131] Exemplary cancers that may be treated according to this embodiment of the invention include, but are not limited to, those described hereinabove.

[0132] In one particular embodiment, the cancer is breast cancer. Suitably, the breast cancer is hormone receptor positive, HER2 negative / low (HR + / HER2-negative / low) breast cancer or triple-negative breast cancer (TNBC).

[0133] Preferably, the breast cancer is hormone receptor positive, HER2 negative / low (HR + / HER2-negative / low) metastatic breast cancer or metastatic triple-negative breast cancer (TNBC).

[0134] Treatment options for one or more subgroups In one embodiment, the subject has one or more of: low monocyte count, luminal B breast cancer, age less than about 85 years, previously treated with a CDK4 / 6 inhibitor, not previously treated with taxane chemotherapy, has an elevated neutrophil to lymphocyte ratio, and was diagnosed less than about 5 years ago.

[0135] In one particular embodiment, the subject has one or more of a low monocyte count, has not previously been treated with taxane chemotherapy, has an elevated neutrophil to lymphocyte ratio, and was diagnosed less than about 5 years ago.

[0136] In another specific embodiment, the subject has previously been treated with a CDK4 / 6 inhibitor and has one or more of a low monocyte count, has not previously been treated with taxane chemotherapy, has an elevated neutrophil to lymphocyte ratio, and was diagnosed less than about 5 years ago.

[0137] In one particular embodiment, the subject has not been previously treated with taxane chemotherapy and has an elevated neutrophil to lymphocyte ratio.

[0138] In another particular embodiment, the subject has been previously treated with a CDK4 / 6 inhibitor, has not previously been treated with taxane chemotherapy, and has an elevated neutrophil to lymphocyte ratio.

[0139] <Therapeutic Methods with High Dose of LAG-3 Protein or Its Derivatives> In a further embodiment, the present invention provides a LAG-3 protein or a derivative thereof capable of binding to an MHC class II molecule for use in preventing, treating or ameliorating cancer in a subject, wherein the LAG-3 protein or derivative thereof is administered to a subject at a dosage of >30 mg to 200 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0140] In a further embodiment, the present invention provides the use of a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment or amelioration of cancer in a subject, wherein the LAG-3 protein or derivative is administered to the subject at a dosage of >30 mg to 200 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0141] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject, comprising administering to the subject a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, at a dosage of >30 mg to 200 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0142] In a further embodiment, the present invention provides a LAG-3 protein or a derivative thereof capable of binding to an MHC class II molecule for use in preventing, treating or ameliorating cancer in a subject, wherein the LAG-3 protein or derivative thereof is administered to a subject at a dosage of >30 mg to 180 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0143] In a further embodiment, the present invention provides the use of a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment or amelioration of cancer in a subject, wherein the LAG-3 protein or derivative is administered to the subject at a dosage of >30 mg to 180 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0144] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject, comprising administering to the subject a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, at a dosage of >30 mg to 180 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0145] In a further embodiment, the present invention provides a LAG-3 protein or a derivative thereof capable of binding to an MHC class II molecule for use in preventing, treating or ameliorating cancer in a subject, wherein the LAG-3 protein or derivative thereof is administered to a subject at a dosage of >30 mg to 150 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0146] In a further embodiment, the present invention provides the use of a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment or amelioration of cancer in a subject, wherein the LAG-3 protein or derivative is administered to the subject at a dosage of >30 mg to 150 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0147] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject, comprising administering to the subject a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, at a dosage of >30 mg to 150 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0148] In a further embodiment, the present invention provides a LAG-3 protein or a derivative thereof capable of binding to an MHC class II molecule for use in preventing, treating or ameliorating cancer in a subject, wherein the LAG-3 protein or derivative thereof is administered to a subject at a dosage of >30 mg to 120 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0149] In a further embodiment, the present invention provides the use of a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment or amelioration of cancer in a subject, wherein the LAG-3 protein or derivative is administered to the subject at a dosage of >30 mg to 120 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0150] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject, comprising administering to the subject a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, at a dosage of >30 mg to 120 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0151] In a further embodiment, the present invention provides a LAG-3 protein or a derivative thereof capable of binding to an MHC class II molecule for use in preventing, treating or ameliorating cancer in a subject, wherein the LAG-3 protein or derivative thereof is administered to a subject at a dosage of >30 mg to <120 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0152] In a further embodiment, the present invention provides the use of a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment or amelioration of cancer in a subject, wherein the LAG-3 protein or derivative is administered to the subject at a dosage of >30 mg to <120 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0153] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject, comprising administering to the subject a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, at a dosage of >30 mg to <120 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0154] Optionally, the LAG-3 protein or derivative thereof is administered to the subject at a dosage of between 40 mg and 200 mg of the molar equivalent of IMP321.

[0155] Optionally, the LAG-3 protein or derivative thereof is administered to the subject at a dosage of between 40 mg and 180 mg of the molar equivalent of IMP321.

[0156] Optionally, the LAG-3 protein or derivative thereof is administered to the subject at a dosage of between 40 mg and 150 mg of the molar equivalent of IMP321.

[0157] Optionally, the LAG-3 protein or derivative thereof is administered to the subject at a dosage of between 40 mg and 120 mg of the molar equivalent of IMP321.

[0158] Optionally, the LAG-3 protein or derivative thereof is administered to the subject at a dosage of between 50 mg and 200 mg of the molar equivalent of IMP321.

[0159] Optionally, the LAG-3 protein or derivative thereof is administered to the subject at a dosage of between 50 mg and 180 mg of the molar equivalent of IMP321.

[0160] Optionally, the LAG-3 protein or derivative thereof is administered to the subject at a dosage of between 50 mg and 150 mg of the molar equivalent of IMP321.

[0161] Optionally, the LAG-3 protein or derivative thereof is administered to the subject at a dosage of between 50 mg and 120 mg of the molar equivalent of IMP321.

[0162] Optionally, the LAG-3 protein or derivative thereof is administered to the subject at a dosage of the molar equivalent of 50-100 mg of IMP321.

[0163] Optionally, the LAG-3 protein or derivative thereof is administered to the subject at a dosage of between 60 mg and 200 mg of the molar equivalent of IMP321.

[0164] Optionally, the LAG-3 protein or derivative thereof is administered to the subject at a dosage of between 60 mg and 180 mg of the molar equivalent of IMP321.

[0165] Optionally, the LAG-3 protein or derivative thereof is administered to the subject at a dosage of between 60 mg and 150 mg of the molar equivalent of IMP321.

[0166] Optionally, the LAG-3 protein or derivative thereof is administered to the subject at a dosage of between 60 mg and 120 mg of the molar equivalent of IMP321.

[0167] Optionally, the LAG-3 protein or derivative thereof is administered to the subject at a dosage of 60-100 mg of IMP321, or the molar equivalent of 60-90 mg of IMP321.

[0168] Optionally, the LAG-3 protein or derivative thereof is administered to the subject at a dosage of the molar equivalent of 80-100 mg of IMP321.

[0169] Optionally, the LAG-3 protein or derivative thereof is administered to the subject at a dosage that is the molar equivalent of 90 mg of IMP321.

[0170] The dosage of LAG-3 protein or its derivative may be administered to a subject in two or more separate doses, each separate dose including a partial dose to provide a total dose in combination.For example, a 90mg dose of IMP321 may be administered in two separate doses of 45mg (e.g., by subcutaneous injection).The separate doses may be separated by up to 30 minutes, for example, up to 15 minutes.

[0171] Optionally, multiple doses of the LAG-3 protein or derivative thereof are administered to the subject.

[0172] Optionally, the LAG-3 protein or derivative thereof is administered to the subject before, during, or after administration of a chemotherapeutic agent.

[0173] Optionally, multiple doses of the chemotherapeutic agent are administered to the subject.

[0174] Optionally, the LAG-3 protein or derivative is administered to the subject on the same day as the chemotherapy agent.

[0175] Optionally, the LAG-3 protein or derivative is administered to the subject on days 1 and 15 of a 4-week cycle, and the chemotherapeutic agent is administered to the subject on days 1, 8, and 15 of the 4-week cycle, and optionally, the 4-week cycle is repeated for 4 to 8 cycles, preferably 6 cycles.

[0176] Optionally, the LAG-3 protein or derivative is administered to the subject on days 1 and 15 of a 4-week cycle, and the chemotherapeutic agent is administered to the subject on days 1, 8, and 15 of the 4-week cycle, and optionally the 4-week cycle is repeated for 4 to 13 cycles, preferably 6, 7, 8, 9, 10, 11, 12 or 13 cycles.

[0177] Optionally, the LAG-3 protein or derivative is administered to the subject one or more times in the absence of a chemotherapeutic agent, after which one or more doses of the LAG-3 protein or derivative are administered to the subject prior to, along with, or following one or more doses of the chemotherapeutic agent.

[0178] Optionally, the LAG-3 protein or derivative is administered to the subject in the absence of a chemotherapeutic agent on days 1 and 15 of a four-week cycle for up to nine four-week cycles.

[0179] Optionally, the LAG-3 protein or derivative is administered to the subject in the absence of a chemotherapeutic agent on days 1 and 15 of a four-week cycle for up to seven four-week cycles.

[0180] Optionally, the LAG-3 protein or derivative is administered to the subject over four to eight, preferably six, four-week cycles, in the absence of the chemotherapy agent following doses of the LAG-3 protein or derivative on days 1 and 15 of the four-week cycle, and doses of the chemotherapy agent on days 1, 8, and 15 of the four-week cycle.

[0181] Optionally, the LAG-3 protein or derivative is administered to the subject for 4 to 13 cycles, preferably 6, 7, 8, 9, 10, 11, 12 or 13 4-week cycles, in the absence of the chemotherapy agent following doses of the LAG-3 protein or derivative on days 1 and 15 of the 4-week cycle, and doses of the chemotherapy agent on days 1, 8 and 15 of the 4-week cycle.

[0182] Optionally, the chemotherapeutic agent is administered at a dosage in accordance with the approved prescribing information.

[0183] Optionally, the chemotherapeutic agent is a taxane.

[0184] Optionally, the chemotherapeutic agent is paclitaxel.

[0185] Optionally, the LAG-3 derivative is the LAG-3Ig fusion protein IMP321.

[0186] Optionally, 80 mg / m 2 of paclitaxel is intravenously administered to the subject on days 1, 8, and 15 of a 4-week cycle, followed by 90 mg of IMP321 being subcutaneously administered to the subject on days 1 and 15 of a 4-week cycle.

[0187] Optionally, the LAG-3 derivative is the LAG-3Ig fusion protein IMP321.

[0188] Optionally, the cancer is breast cancer.

[0189] Optionally, the cancer is metastatic breast cancer.

[0190] Optionally, the subject is a hormone receptor-positive HER2-negative / low (HR + / HER2-negative / low) metastatic breast cancer patient, or a metastatic triple-negative breast cancer (TNBC) patient.

[0191] <Method of treating on the same day with an LAG-3 protein or its derivative and a chemotherapeutic agent> In a further embodiment, the present invention provides an LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for use in the prevention, treatment, or amelioration of cancer in a subject, wherein the LAG-3 protein or derivative is administered to the subject on the same day as a chemotherapeutic agent.

[0192] In a further embodiment, the present invention provides the use of an LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject, wherein the LAG-3 protein or derivative is administered to the subject on the same day as a chemotherapeutic agent.

[0193] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject, comprising administering to the subject a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, on the same day as a chemotherapy agent.

[0194] Optionally, the LAG-3 protein or derivative is administered to the subject on days 1 and 15 of a 4-week cycle, and the chemotherapeutic agent is administered to the subject on days 1, 8, and 15 of the 4-week cycle, and optionally, the 4-week cycle is repeated for 4 to 8 cycles, preferably 6 cycles.

[0195] Optionally, the LAG-3 protein or derivative is administered at a dosage of 6 mg to <120 mg, preferably 20 mg to 100 mg, more preferably 30 mg to 90 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0196] Optionally, the chemotherapeutic agent is administered at a dosage in accordance with the approved prescribing information.

[0197] Optionally, the chemotherapeutic agent is a taxane.

[0198] Optionally, the chemotherapeutic agent is paclitaxel.

[0199] Optionally, the LAG-3 derivative is a LAG-3Ig fusion protein, IMP321.

[0200] Optionally, 80 mg / m 2 of paclitaxel will be administered intravenously to subjects on days 1, 8, and 15 of a 4-week cycle, followed by 90 mg of IMP321 administered subcutaneously on days 1 and 15 of a 4-week cycle.

[0201] Optionally, the cancer is breast cancer.

[0202] Optionally, the cancer is metastatic breast cancer.

[0203] Optionally, the subject is a hormone receptor positive HER2-negative / low (HR + / HER2-negative / low) metastatic breast cancer patient, or a metastatic triple-negative breast cancer (TNBC) patient.

[0204] <Method of treatment with an LAG-3 protein or derivative thereof in the absence of a chemotherapeutic agent following combination treatment with the LAG-3 protein or derivative thereof and a chemotherapeutic agent> In a further embodiment, the present invention provides an LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for use in the prevention, treatment, or amelioration of cancer in a subject, wherein the LAG-3 protein or derivative is administered to the subject one or more times in the absence of a chemotherapeutic agent after one or more dosages of the LAG-3 protein or derivative have been administered to the subject before, with, or after one or more dosages of the chemotherapeutic agent.

[0205] In a further embodiment, the present invention provides the use of an LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject, wherein the LAG-3 protein or derivative is administered to the subject one or more times in the absence of a chemotherapeutic agent after one or more dosages of the LAG-3 protein or derivative have been administered to the subject before, with, or after one or more dosages of the chemotherapeutic agent.

[0206] In a further embodiment, the present invention provides a method for the prevention, treatment, or amelioration of cancer in a subject, comprising administering to the subject one or more times an LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the absence of a chemotherapeutic agent after one or more dosages of the LAG-3 protein or derivative have been administered to the subject before, with, or after one or more dosages of the chemotherapeutic agent.

[0207] Optionally, the LAG-3 protein or derivative is administered to the subject in the absence of a chemotherapeutic agent on days 1 and 15 of a four-week cycle for up to nine four-week cycles.

[0208] Optionally, the LAG-3 protein or derivative is administered to the subject in the absence of a chemotherapeutic agent on days 1 and 15 of a four-week cycle for up to seven four-week cycles.

[0209] Optionally, the LAG-3 protein or derivative is administered to the subject over four to eight, preferably six, four-week cycles, in the absence of the chemotherapy agent following doses of the LAG-3 protein or derivative on days 1 and 15 of the four-week cycle, and doses of the chemotherapy agent on days 1, 8, and 15 of the four-week cycle.

[0210] Optionally, the LAG-3 protein or derivative is administered to the subject over 4 to 13, preferably 6, 7, 8, 9, 10, 11, 12 or 13, four-week cycles, in the absence of the chemotherapy agent following doses of the LAG-3 protein or derivative on days 1 and 15 of the four-week cycle, and doses of the chemotherapy agent on days 1, 8 and 15 of the four-week cycle.

[0211] Optionally, the LAG-3 protein or derivative is administered at a dosage of 1 mg to 200 mg, 6 mg to 200 mg, 6 mg to <120 mg, 20 mg to 100 mg, 30 mg to 90 mg, about 30 mg, or about 90 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0212] Optionally, the chemotherapeutic agent is administered at a dosage in accordance with the approved prescribing information.

[0213] Optionally, the chemotherapeutic agent is a taxane.

[0214] Optionally, the chemotherapeutic agent is paclitaxel.

[0215] Optionally, the LAG-3 derivative is a LAG-3Ig fusion protein, IMP321.

[0216] Optionally, 80 mg / m 2 of paclitaxel will be administered intravenously to subjects on days 1, 8, and 15 of a 4-week cycle, followed by 90 mg of IMP321 administered subcutaneously on days 1 and 15 of a 4-week cycle.

[0217] Optionally, the cancer is breast cancer.

[0218] Optionally, the cancer is metastatic breast cancer.

[0219] Optionally, the subject is a hormone receptor positive HER2-negative / low (HR + Patients with HER2-negative / low) metastatic breast cancer or metastatic triple-negative breast cancer (TNBC).

[0220] Treatment of patient subgroups In a further embodiment, the present invention relates to a LAG-3 protein or a derivative thereof capable of binding to an MHC class II molecule for use in the prevention, treatment or amelioration of metastatic breast cancer in a subject, the subject being hormone receptor positive, HER2-negative / low (HR + The present invention provides a LAG-3 protein or derivative thereof for use in patients with HER2-negative / low (HER2-negative / low) metastatic breast cancer, or metastatic triple-negative breast cancer (TNBC) patients.

[0221] In a further embodiment, the present invention relates to the use of a LAG-3 protein or a derivative thereof capable of binding to an MHC class II molecule in the manufacture of a medicament for the prevention, treatment or amelioration of metastatic breast cancer in a subject, the subject being hormone receptor positive, HER2-negative / low (HR + The present invention provides for use in patients with HER2-negative / low (HER2-negative / low) metastatic breast cancer or metastatic triple-negative breast cancer (TNBC).

[0222] In a further embodiment, the present invention is a method for preventing, treating, or ameliorating metastatic breast cancer in a subject, comprising administering to the subject an LAG-3 protein, or a derivative thereof that can bind to an MHC class II molecule, wherein the subject is a hormone receptor positive HER2-negative / low (HR + / HER2-negative / low) metastatic breast cancer patient, or a metastatic triple negative breast cancer (TNBC) patient.

[0223] <LAG-3 Protein and Derivatives> According to an embodiment of the present invention, the LAG-3 protein may be an isolated native or recombinant LAG-3 protein. The LAG-3 protein may comprise the amino acid sequence of an LAG-3 protein from any suitable species, such as a primate or mouse LAG-3 protein, but is preferably the amino acid sequence of a human LAG-3 protein. The amino acid sequences of human and mouse LAG-3 proteins are shown in FIG. 1 of Huard et al. (Proc. Natl. Acad. Sci. USA, 11:5744-5749, 1997). The sequence of the human LAG-3 protein is repeated in FIG. 1 herein (SEQ ID NO: 1). The amino acid sequences of the four extracellular Ig superfamily domains (D1, D2, D3, and D4) of human LAG-3 are also identified by the amino acid residues: 1-149 (D1), 150-239 (D2), 240-330 (D3), and 331-412 (D4) of FIG. 1 of Huard et al.

[0224] Derivatives of the LAG-3 protein include soluble fragments, mutants, or variants of the LAG-3 protein that are capable of binding to MHC class II molecules. Several derivatives of the LAG-3 protein are known to be capable of binding to MHC class II molecules. Many examples of such derivatives can be determined using quantitative cell adhesion assays such as those described in Huard et al. (Proc. Natl. Acad. Sci. USA, 11:5744-5749, 1997). This document describes the characterization of the MHC class II binding site on the LAG-3 protein. Methods for making mutants of LAG-3 are described, as well as quantitative cell adhesion assays to determine the ability of LAG-3 mutants to bind to class II positive Daudi cells. The binding of several different mutants of LAG-3 to MHC class II molecules was determined. Some mutants were able to reduce class II binding, while others increased the affinity of LAG-3 for class II molecules. Many of the residues essential for LAG-3 binding to MHC class II proteins are concentrated at the base of a large 30 amino acid extra loop structure in the D1 domain of LAG-3. The amino acid sequence of the extra loop structure of the D1 domain of human LAG-3 protein is GPPAAAPGHPLAPGPHPAAPSSWGPRPRRY (SEQ ID NO: 2). The amino acid sequence of the extra loop structure of the D1 domain of human LAG-3 protein is shown in bold underline in Figure 1.

[0225] In one embodiment of the present invention, the derivative of the LAG-3 protein comprises the 30 amino acid extra loop sequence of the D1 domain of human LAG-3, or a variant of such a sequence having one or more amino acid substitutions. The variant may comprise an amino acid sequence having at least 70%, 80%, 90%, or 95% amino acid identity with the 30 amino acid extra loop sequence of the D1 domain of human LAG-3. Optionally, the amino acid substitution is a conservative amino acid substitution.

[0226] A derivative of a LAG-3 protein may comprise the amino acid sequence of domains D1 and optionally D2, or domains D1 and D2, of a LAG-3 protein, preferably a human LAG-3 protein.

[0227] A derivative of a LAG-3 protein may comprise an amino acid sequence having at least 70%, 80%, 90%, or 95% amino acid identity to domains D1 and optionally D2, or domains D1 and D2, of a LAG-3 protein, preferably a human LAG-3 protein.

[0228] A derivative of a LAG-3 protein may comprise the amino acid sequence of domains D1, D2, and D3, domains D1, D2, D3 and optionally D4, or domains D1, D2, D3 and D4 of a LAG-3 protein, preferably a human LAG-3 protein.

[0229] A derivative of a LAG-3 protein may comprise an amino acid sequence having at least 70%, 80%, 90%, or 95% amino acid identity to a LAG-3 protein, preferably domains D1, D2 and D3, domains D1, D2, D3 and optionally D4, or domains D1, D2, D3 and D4 of human LAG-3.

[0230] The sequence identity between amino acid sequences can be determined by comparing the alignment of sequences. If the corresponding positions in the compared sequences are occupied by the same amino acid, the molecules are identical at that position. The score of the alignment as a percentage of identity is a function of the number of identical amino acids at the positions shared by the compared sequences. When comparing sequences, optimal alignment may require that gaps are introduced in one or more sequences to take into account possible insertions and deletions in the sequence. The sequence comparison method may use a gap penalty for the same number of identical molecules in the compared sequences, and a sequence alignment with as few gaps as possible will achieve a higher score than one with many gaps, reflecting a higher relatedness between the two sequences being compared. The calculation of the maximum identity percentage includes the generation of an optimal alignment that takes into account gap penalties.

[0231] Suitable computer programs for carrying out sequence comparisons are widely available in the commercial and public sector. Examples include MatGat (Campanella et al., 2003, BMC Bioinformatics 4:29; program available at http: / / bitincka.com / ledion / matgat), Gap (Needleman and Wunsch, 1970, J. Mol. Biol. 48:443-453), FASTA (Altschul et al., 1990, J. Mol. Biol. 215:403-410; program available at http: / / www.ebi.ac.uk / fasta), Clustal W 2.0 and X 2.0 (Larkin et al., 2007, Bioinformatics 23:2947-2948; program available at http: / / www.ebi.ac.uk / tools / clustalw2) and EMBOSS Pairwise Alignment. Algorithms (Needleman and Wunsch, 1970, supra; Kruskal, 1983, Time warps, string edits and macromolecules: the theory and practice of sequence comparison, Sankoff and Kruskal (eds.), pp. 1-44, Addison Wesley; programs available at http: / / www.ebi.ac.uk / tools / emboss / align). All programs can be run using the default parameters.

[0232] For example, sequence comparison can be performed using the "needle" method of the EMBOSS Pairwise Alignment Algorithms, which determines the optimal alignment (including gaps) when considering two sequences over their entire length and provides a percentage identity score. Default parameters for amino acid sequence comparison (option "protein molecule") can be Gap Extend penalty: 0.5, Gap Open penalty: 10.0, Matrix: Blosum 62.

[0233] Sequence comparison may be performed over the entire length of the reference sequence.

[0234] Derivatives of the LAG-3 protein may be fused to an immunoglobulin Fc amino acid sequence, preferably a human IgG1 Fc amino acid sequence, optionally by a linker amino acid sequence.

[0235] The ability of a derivative of the LAG-3 protein to bind to an MHC class II molecule may be determined using a quantitative cell adhesion assay such as that described in Huard et al. (Proc. Natl. Acad. Sci. USA, 11:5744-5749, 1997). The affinity of the derivative of the LAG-3 protein for an MHC class II molecule may be at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% of the affinity of the human LAG-3 protein for an MHC class II molecule.

[0236] Preferably, the affinity of the LAG-3 protein derivative for MHC class II molecules is at least 50%, 60%, 70%, 80%, 90%, 95%, 99%, or 100% of the affinity of human LAG-3 protein for MHC class II molecules.

[0237] Examples of suitable derivatives of the LAG-3 protein capable of binding to MHC class II molecules include: amino acid residues 23–448 of the human LAG-3 sequence; Amino acid sequences of domains D1 and D2 of LAG-3; The amino acid sequences of domains D1 and D2 of LAG-3 with amino acid substitutions at one or more of the following positions: position 30 where ASP is replaced by ALA; position 56 where HIS is replaced by ALA; position 73 where ARG is replaced by GLU; position 75 where ARG is replaced by ALA or GLU; position 76 where ARG is replaced by GLU; or position 103 where ARG is replaced by ALA; Derivatives include: recombinant soluble human LAG-3Ig fusion protein (IMP321)-160 kDa dimer produced in Chinese hamster ovary cells transfected with a plasmid encoding the extracellular domain of hLAG-3 fused to human IgG1 Fc. The sequence of IMP321 is given in SEQ ID NO: 17 of US Patent Application Publication No. 2011 / 0008331.

[0238] In one embodiment, the subject is a mammal, preferably a human.

[0239] According to the present invention, LAG-3 protein or its derivative is administered in a therapeutically effective amount. "Therapeutically effective amount" refers to the amount of active ingredient that is sufficient to have a therapeutic effect when administered. The effective amount of active ingredient varies, for example, according to the specific disease or diseases to be treated, the severity of the disease, the duration of treatment, and the characteristics of the patient (for example, sex, age, height and weight).

[0240] In one embodiment, the LAG-3 protein or derivative thereof is administered at a dose that is the molar equivalent of about 0.1 mg to about 200 mg, about 1 mg to about 200 mg, about 6 mg to about 200 mg, about 10 mg to about 200 mg, about 20 mg to about 200 mg, about 30 mg to about 200 mg, >30 mg to about 200 mg, about 40 mg to about 200 mg, about 50 mg to about 200 mg, or about 80 mg to about 200 mg of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0241] In one embodiment, the LAG-3 protein or derivative thereof is administered at a dose that is the molar equivalent of about 0.1 mg to about 180 mg, about 1 mg to about 180 mg, about 6 mg to about 180 mg, about 10 mg to about 180 mg, about 20 mg to about 180 mg, about 30 mg to about 180 mg, >30 mg to about 180 mg, about 40 mg to about 180 mg, about 50 mg to about 180 mg, or about 80 mg to about 180 mg of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0242] In one embodiment, the LAG-3 protein or derivative thereof is administered at a dose that is the molar equivalent of about 0.1 mg to about 120 mg, about 1 mg to about 120 mg, about 6 mg to about 120 mg, about 10 mg to about 120 mg, about 20 mg to about 120 mg, about 30 mg to about 120 mg, >30 mg to about 120 mg, about 40 mg to about 120 mg, about 50 mg to about 120 mg, or about 80 mg to about 120 mg of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0243] In one embodiment, the LAG-3 protein or derivative thereof is administered at a dose that is the molar equivalent of about 0.1 mg to about 60 mg, about 6 mg to about 60 mg, about 10 mg to about 50 mg, about 20 mg to about 40 mg, about 25 mg to about 35 mg, or about 30 mg of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0244] In another embodiment, the LAG-3 protein or derivative thereof is administered at a dose that is the molar equivalent of about 25 mg, about 26 mg, about 27 mg, about 28 mg, about 29 mg, about 30 mg, about 31 mg, about 32 mg, about 33 mg, about 34 mg, or about 35 mg of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0245] Suitably, the LAG-3 protein or derivative thereof is administered in a dose that is the molar equivalent of about 30 mg of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0246] In yet another embodiment, the LAG-3 protein or derivative thereof is administered at a dose that is the molar equivalent of about 25 mg to about 60 mg, e.g., about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, or about 60 mg of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0247] In one embodiment, the LAG-3 protein or derivative thereof is IMP321 and is administered at a dose of about 0.1 mg to about 60 mg, about 6 mg to about 60 mg, about 10 mg to about 50 mg, about 20 mg to about 40 mg, about 25 mg to about 35 mg, or about 30 mg.

[0248] In another embodiment, IMP321 is administered at a dose of about 25 mg, about 26 mg, about 27 mg, about 28 mg, about 29 mg, about 30 mg, about 31 mg, about 32 mg, about 33 mg, about 34 mg, or about 35 mg.

[0249] Suitably, IMP321 is administered in a dose of about 30 mg.

[0250] In other embodiments, IMP321 is administered at a dose of about 25 mg to about 60 mg, for example, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, or about 60 mg.

[0251] Doses of 6-30 mg per subcutaneous (sc) injection of IMP321 have thus far been shown to be safe and provide acceptable systemic exposure based on the results of pharmacokinetic data obtained in patients with metastatic renal cell carcinoma. Blood concentrations of IMP321 greater than 1 ng / ml are achieved for at least 24 hours after sc injection in patients injected with doses of IMP321 greater than 6 mg. No dose-limiting toxicities have been observed to date.

[0252] In one embodiment, the LAG-3 protein or derivative thereof is administered to the subject about once a week. In another embodiment, the LAG-3 protein or derivative thereof is administered to the subject about once every two weeks. In yet another embodiment, the LAG-3 protein or derivative thereof is administered to the subject about once every three weeks. In a further embodiment, the LAG-3 protein or derivative thereof is administered to the subject about once every four weeks. In yet a further embodiment, the LAG-3 protein or derivative thereof is administered to the subject about once a month. As will be appreciated by those skilled in the art, the exact treatment regimen will vary and be adapted according to the particular cancer being treated and the characteristics of the patient.

[0253] In one embodiment, the LAG-3 protein or derivative thereof is present as the sole active ingredient, hi another embodiment, the LAG-3 protein or derivative thereof is present in the absence of any further antigens added to the pharmaceutical composition or medicament.

[0254] <Combination treatment with chemotherapy in one or more subgroups> In one embodiment, the LAG-3 protein or derivative thereof is administered in combination with a chemotherapeutic agent.

[0255] Suitable chemotherapeutic agents include, but are not limited to, alkylating agents, plant alkaloids, antitumor antibiotics, antimetabolites, topoisomerase inhibitors, and miscellaneous antitumor drugs.

[0256] Preferably, the chemotherapeutic agent is an alkylating agent. Exemplary alkylating agents include mustard gas derivatives such as mechlorethamine, cyclophosphamide, chlorambucil, melphalan, and ifosfamide; ethyleneimines such as thiotepa and hexamethylmelamine; alkylsulfonates such as busulfan; hydrazines and triazines such as altretamine, procarbazine, dacarbazine, and temozolomide; nitrosoureas such as carmustine, lomustine, and streptozocin; and metal salts such as carboplatin, cisplatin, and oxaliplatin.

[0257] Preferably, the chemotherapeutic agent is a plant alkaloid.Exemplary plant alkaloids include vinca alkaloids such as vincristine, vinblastine and vinorelbine; taxanes such as paclitaxel, nab-paclitaxel, docetaxel, cabazitaxel, larotaxel, mirataxel, ortataxel, taxoplexin, opaxio, tesetaxel and BMS-184476; podophyllotoxins such as etoposide and tenisopide; and camptothecan analogs such as irinotecan and topotecan.

[0258] Preferably, the chemotherapeutic agent is an antitumor antibiotic. Exemplary antitumor antibiotics include anthracyclines such as doxorubicin, daunorubicin, epirubicin, mitoxantrone, and idarubicin; chromomycins such as dactinomycin and plicamycin; and various antitumor antibiotics such as mitomycin and bleomycin.

[0259] Preferably, the chemotherapeutic agent is an antimetabolite. Exemplary antimetabolites include folate antagonists such as methotrexate, pyrimidine antagonists such as 5-fluorouracil, foxuridine, cytarabine, capecitabine and gemcitabine, purine antagonists such as 6-mercaptopurine and 6-thioguanine, and adenosine deaminase inhibitors such as cladribine, fludarabine, nelarabine and pentostatin.

[0260] Preferably, the chemotherapeutic agent is a topoisomerase inhibitor. Exemplary topoisomerase inhibitors include topoisomerase I inhibitors, such as irinotecan and topotecan; and topoisomerase II inhibitors, such as amsacrine, etoposide, etoposide phosphate, and teniposide.

[0261] Preferably, the chemotherapeutic agents are various anti-tumor drugs.

[0262] Exemplary anti-tumor drugs include ribonucleotide reductase inhibitors such as hydroxyurea; corticosteroid inhibitors such as mitotane; enzymes such as asparaginase and pegaspargase; microtubule inhibitors such as estramustine; and retinoids such as bexarotene, isotretinoin, and tretinoin.

[0263] In one particular embodiment, the chemotherapeutic agent is a taxane. In one embodiment, the taxane is paclitaxel, nab-paclitaxel, docetaxel, cabazitaxel, larotaxel, mirataxel, ortataxel, taxoplexin, opaxio, tesetaxel, or BMS-184476.

[0264] In another embodiment, the taxane is paclitaxel.

[0265] The chemotherapeutic agent is administered in a therapeutically effective amount. The therapeutically effective amount refers to the amount of the chemotherapeutic agent that is sufficient to have a therapeutic effect when administered. The effective amount of the chemotherapeutic agent varies depending on the chemotherapeutic agent selected, the specific disease or diseases being treated, the severity of the disease, the duration of the treatment, and the characteristics of the patient (e.g., sex, age, height and weight). Preferably, the chemotherapeutic agent is administered according to the prescription information approved for the chemotherapeutic agent.

[0266] In some embodiments, the chemotherapeutic agents are administered parenterally (including by subcutaneous, intravenous or intramuscular injection) or orally. Preferably, the chemotherapy is administered intravenously.

[0267] In one embodiment, the LAG-3 protein or derivative thereof is administered before, during or after administration of the chemotherapeutic agent, hi another embodiment, the LAG-3 protein or derivative thereof is administered after administration of the chemotherapeutic agent.

[0268] In one embodiment, the LAG-3 protein or derivative thereof and the chemotherapeutic agent are packaged separately, i.e., in this embodiment, the LAG-3 protein or derivative thereof and the chemotherapeutic agent are in separate unit dosage forms, typically (but not necessarily) supplied by different suppliers, and then used in the methods of the invention.

[0269] In another embodiment, the LAG-3 protein or derivative thereof and the chemotherapeutic agent are in the form of a combined formulation.

[0270] The components of a "combination preparation" may be present (i) in one combination unit dosage form, known as a fixed dose combination (FDC), or (ii) as a first unit dosage form of component (a) and a separate second unit dosage form of component (b) packaged together, known as a kit of parts. The ratio of the total amounts of combination component (a) to combination component (b) administered in a combination preparation may vary, for example, to accommodate the needs of a patient subpopulation being treated, or the needs of a patient, which may result from, for example, a particular disease, age, sex, or weight of the patient.

[0271] That is, the combination preparation according to the present invention may take the form of a pharmaceutical composition comprising the LAG-3 protein or a derivative thereof and a chemotherapeutic agent, or it may take the form of a kit-of-parts comprising the LAG-3 protein or a derivative thereof and a chemotherapeutic agent as separate components but packaged together.

[0272] The kit-of-parts may include multiple doses of the LAG-3 protein or derivative thereof and / or multiple doses of a chemotherapeutic agent.

[0273] In one embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent for use in the prevention, treatment, or amelioration of cancer in a subject with low monocyte counts.

[0274] In another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject with low monocyte count.

[0275] In yet another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent for the prevention, treatment, or amelioration of cancer in a subject with low monocyte count.

[0276] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject with low monocyte counts, comprising administering to a subject in need of such prevention, treatment or amelioration a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent.

[0277] In yet a further embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for use in the prevention, treatment or amelioration of cancer in a subject with low monocyte count, wherein the LAG-3 protein, or a derivative thereof, is administered before, during or after administration of a chemotherapeutic agent.

[0278] In another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject with low monocyte count, wherein the LAG-3 protein, or a derivative thereof, is administered before, during or after administration of a chemotherapeutic agent.

[0279] In one embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject with low monocyte counts, comprising administering to a subject in need of such prevention, treatment or amelioration a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent, wherein the LAG-3 protein or derivative thereof is administered before, during or after administration of the chemotherapeutic agent.

[0280] In one embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent for use in the prevention, treatment, or amelioration of luminal B breast cancer in a subject.

[0281] In another embodiment, the present invention relates to the use of a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent in the manufacture of a medicament for the prevention, treatment, or amelioration of luminal B breast cancer in a subject.

[0282] In yet another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent for the prevention, treatment, or amelioration of luminal B breast cancer in a subject.

[0283] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating luminal B breast cancer in a subject, comprising administering to a subject in need of such prevention, treatment or amelioration a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent.

[0284] In yet a further embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for use in preventing, treating, or ameliorating luminal B breast cancer in a subject, wherein the LAG-3 protein, or a derivative thereof, is administered before, during or after administration of a chemotherapeutic agent.

[0285] In another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment, or amelioration of luminal B breast cancer in a subject, wherein the LAG-3 protein, or a derivative thereof, is administered before, during or after administration of a chemotherapeutic agent.

[0286] In one embodiment, the present invention provides a method for preventing, treating or ameliorating luminal B breast cancer in a subject, comprising administering to a subject in need of such prevention, treatment or amelioration a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent, wherein the LAG-3 protein or derivative thereof is administered before, during or after administration of the chemotherapeutic agent.

[0287] In one embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent for use in the prevention, treatment, or amelioration of cancer in a subject less than about 85 years of age.

[0288] In another embodiment, the present invention relates to the use of a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject less than about 85 years of age.

[0289] In yet another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent for the prevention, treatment, or amelioration of cancer in a subject less than about 85 years of age.

[0290] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject less than about 85 years of age, comprising administering to a subject in need of such prevention, treatment or amelioration a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent.

[0291] In yet a further embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for use in the prevention, treatment, or amelioration of cancer in a subject less than about 85 years of age, wherein the LAG-3 protein, or a derivative thereof, is administered before, during or after administration of a chemotherapeutic agent.

[0292] In another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject less than about 85 years of age, wherein the LAG-3 protein, or a derivative thereof, is administered before, during or after administration of a chemotherapeutic agent.

[0293] In one embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject less than about 85 years of age, comprising administering to a subject in need of such prevention, treatment or amelioration a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent, wherein the LAG-3 protein or derivative thereof is administered before, during or after administration of the chemotherapeutic agent.

[0294] In one embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent for use in the prevention, treatment, or amelioration of cancer in a subject previously treated with a CDK4 / 6 inhibitor.

[0295] In another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject previously treated with a CDK4 / 6 inhibitor.

[0296] In another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent for the prevention, treatment, or amelioration of cancer in a subject previously treated with a CDK4 / 6 inhibitor.

[0297] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject previously treated with a CDK4 / 6 inhibitor, comprising administering to a subject in need of such prevention, treatment or amelioration a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent.

[0298] In yet a further embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for use in the prevention, treatment, or amelioration of cancer in a subject previously treated with a CDK4 / 6 inhibitor, wherein the LAG-3 protein, or a derivative thereof, is administered before, during or after administration of a chemotherapeutic agent.

[0299] In another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject previously treated with a CDK4 / 6 inhibitor, wherein the LAG-3 protein, or a derivative thereof, is administered before, during or after administration of a chemotherapeutic agent.

[0300] In one embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject previously treated with a CDK4 / 6 inhibitor, comprising administering to a subject in need of such prevention, treatment or amelioration a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent, wherein the LAG-3 protein or derivative thereof is administered before, during or after administration of the chemotherapeutic agent.

[0301] In one embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent for use in the prevention, treatment, or amelioration of cancer in a subject who has not previously been treated with taxane chemotherapy.

[0302] In another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject who has not previously been treated with taxane chemotherapy.

[0303] In yet another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent for the prevention, treatment, or amelioration of cancer in a subject who has not previously been treated with taxane chemotherapy.

[0304] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject who has not previously been treated with taxane chemotherapy, comprising administering to a subject in need of such prevention, treatment or amelioration a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent.

[0305] In yet a further embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for use in the prevention, treatment or amelioration of cancer in a subject not previously treated with taxane chemotherapy, wherein the LAG-3 protein, or a derivative thereof, is administered before, during or after administration of a chemotherapeutic agent.

[0306] In another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment or amelioration of cancer in a subject who has not previously been treated with taxane chemotherapy, wherein the LAG-3 protein, or a derivative thereof, is administered before, during or after administration of a chemotherapeutic agent.

[0307] In one embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject who has not previously been treated with taxane chemotherapy, comprising administering to a subject in need of such prevention, treatment or amelioration a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent, wherein the LAG-3 protein or derivative thereof is administered before, during or after administration of the chemotherapeutic agent.

[0308] In one embodiment, the present invention relates to LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent for use in the prevention, treatment, or amelioration of cancer in a subject with an elevated neutrophil to lymphocyte ratio.

[0309] In another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject with an elevated neutrophil to lymphocyte ratio.

[0310] In yet another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent for the prevention, treatment, or amelioration of cancer in a subject with an elevated neutrophil-to-lymphocyte ratio.

[0311] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject having an elevated neutrophil to lymphocyte ratio, comprising administering to a subject in need of such prevention, treatment or amelioration a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent.

[0312] In yet a further embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for use in the prevention, treatment, or amelioration of cancer in a subject with an elevated neutrophil-to-lymphocyte ratio, wherein the LAG-3 protein, or a derivative thereof, is administered before, during or after administration of a chemotherapeutic agent.

[0313] In another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject with an elevated neutrophil-to-lymphocyte ratio, wherein the LAG-3 protein, or a derivative thereof, is administered before, during or after administration of a chemotherapeutic agent.

[0314] In one embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject having an elevated neutrophil to lymphocyte ratio, comprising administering to a subject in need of such prevention, treatment or amelioration a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent, wherein the LAG-3 protein or derivative thereof is administered before, during or after administration of the chemotherapeutic agent.

[0315] In one embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent for use in the prevention, treatment, or amelioration of cancer in a subject diagnosed less than about 5 years ago.

[0316] In another embodiment, the present invention relates to the use of a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject diagnosed less than about 5 years ago.

[0317] In yet another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent for the prevention, treatment, or amelioration of cancer in a subject diagnosed less than about 5 years ago.

[0318] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject diagnosed less than about 5 years ago, comprising administering to a subject in need of such prevention, treatment or amelioration a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent.

[0319] In yet a further embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for use in the prevention, treatment, or amelioration of cancer in a subject diagnosed less than about 5 years ago, wherein the LAG-3 protein, or a derivative thereof, is administered before, during or after administration of a chemotherapeutic agent.

[0320] In another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject diagnosed less than about 5 years ago, wherein the LAG-3 protein, or a derivative thereof, is administered before, during or after administration of a chemotherapeutic agent.

[0321] In one embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject diagnosed less than about 5 years ago, comprising administering to a subject in need of such prevention, treatment or amelioration a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent, wherein the LAG-3 protein or derivative thereof is administered before, during or after administration of the chemotherapeutic agent.

[0322] In one embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent for use in the prevention, treatment, or amelioration of cancer in a subject, wherein the subject has one or more of: low monocyte count, luminal B breast cancer, age less than about 85 years, previously treated with a CDK4 / 6 inhibitor, not previously treated with taxane chemotherapy, elevated neutrophil to lymphocyte ratio, and diagnosed less than about 5 years ago.

[0323] In another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject, wherein the subject has one or more of: low monocyte count, luminal B breast cancer, age less than about 85 years, previously treated with a CDK4 / 6 inhibitor, not previously treated with taxane chemotherapy, elevated neutrophil to lymphocyte ratio, and diagnosed less than about 5 years ago.

[0324] In yet another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent for the prevention, treatment, or amelioration of cancer in a subject, wherein the subject has one or more of: low monocyte count, luminal B breast cancer, age less than about 85 years, previously treated with a CDK4 / 6 inhibitor, not previously treated with taxane chemotherapy, elevated neutrophil to lymphocyte ratio, and diagnosed less than about 5 years ago.

[0325] In a further embodiment, the present invention provides a method of preventing, treating or ameliorating cancer in a subject, comprising administering to a subject in need of such prevention, treatment or amelioration a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent, wherein the subject has one or more of: low monocyte count, luminal B breast cancer, age less than about 85 years, previously treated with a CDK4 / 6 inhibitor, not previously treated with taxane chemotherapy, elevated neutrophil to lymphocyte ratio, and diagnosed less than about 5 years ago.

[0326] In yet a further embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for use in preventing, treating, or ameliorating cancer in a subject, wherein the LAG-3 protein or derivative thereof is administered before, during or after administration of a chemotherapeutic agent, and the subject has one or more of: low monocyte count, luminal B breast cancer, age less than about 85 years, previously treated with a CDK4 / 6 inhibitor and not previously treated with taxane chemotherapy, elevated neutrophil to lymphocyte ratio, and diagnosed less than about 5 years ago.

[0327] In another embodiment, the present invention relates to the use of LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject, wherein the LAG-3 protein or derivative thereof is administered before, during or after administration of a chemotherapeutic agent, and the subject has one or more of: low monocyte count, luminal B breast cancer, age less than about 85 years, previously treated with a CDK4 / 6 inhibitor and not previously treated with taxane chemotherapy, elevated neutrophil to lymphocyte ratio, and diagnosed less than about 5 years ago.

[0328] In one embodiment, the present invention provides a method of preventing, treating, or ameliorating cancer in a subject, comprising administering to a subject in need of such prevention, treatment, or amelioration a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent, wherein the LAG-3 protein or derivative thereof is administered before, during, or after administration of the chemotherapeutic agent, and the subject has one or more of the following: low monocyte count, luminal B breast cancer, age less than about 85 years, previously treated with a CDK4 / 6 inhibitor, not previously treated with taxane chemotherapy, elevated neutrophil to lymphocyte ratio, and diagnosed less than about 5 years ago.

[0329] In one particular embodiment, the subject has one or more of a low monocyte count, has not previously been treated with taxane chemotherapy, has an elevated neutrophil to lymphocyte ratio, and was diagnosed less than about 5 years ago.

[0330] In another specific embodiment, the subject has previously been treated with a CDK4 / 6 inhibitor and has one or more of a low monocyte count, has not previously been treated with taxane chemotherapy, has an elevated neutrophil to lymphocyte ratio, and was diagnosed less than about 5 years ago.

[0331] In one particular embodiment, the subject has not been previously treated with taxane chemotherapy and has an elevated neutrophil to lymphocyte ratio.

[0332] In another particular embodiment, the subject has been previously treated with a CDK4 / 6 inhibitor, has not previously been treated with taxane chemotherapy, and has an elevated neutrophil to lymphocyte ratio.

[0333] In a further embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent for use in the prevention, treatment, or amelioration of cancer in a subject, wherein the LAG-3 protein or derivative is administered to the subject at a dosage of >30 mg to <200 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0334] In a further embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent for use in the prevention, treatment, or amelioration of cancer in a subject, wherein the LAG-3 protein or derivative is administered to the subject at a dosage of >30 mg to <120 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0335] In another embodiment, the present invention relates to the use of a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent in the manufacture of a medicament for use in the prevention, treatment, or amelioration of cancer in a subject, wherein the LAG-3 protein or derivative is administered to the subject at a dosage of >30 mg to <200 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0336] In another embodiment, the present invention relates to the use of a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent in the manufacture of a medicament for use in the prevention, treatment, or amelioration of cancer in a subject, wherein the LAG-3 protein or derivative is administered to the subject at a dosage of >30 mg to <120 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0337] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject comprising administering to a subject in need of such prevention, treatment or amelioration a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent, wherein the LAG-3 protein or derivative thereof is administered to the subject at a dosage of >30 mg to <200 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0338] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject comprising administering to a subject in need of such prevention, treatment or amelioration a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent, wherein the LAG-3 protein or derivative thereof is administered to the subject at a dosage of >30 mg to <120 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0339] In yet a further embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for use in preventing, treating, or ameliorating cancer in a subject, wherein the LAG-3 protein or derivative is administered before, during, or after administration of a chemotherapeutic agent, and the LAG-3 protein or derivative is administered to the subject at a dosage of >30 mg to <200 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0340] In yet a further embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for use in preventing, treating, or ameliorating cancer in a subject, wherein the LAG-3 protein or derivative is administered before, during, or after administration of a chemotherapeutic agent, and the LAG-3 protein or derivative is administered to the subject at a dosage of >30 mg to <120 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0341] In another embodiment, the present invention relates to the use of a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject, wherein the LAG-3 protein or derivative is administered before, during, or after administration of a chemotherapeutic agent, and the LAG-3 protein or derivative is administered to the subject at a dosage of >30 mg to <200 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0342] In another embodiment, the present invention relates to the use of a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment or amelioration of cancer in a subject, wherein the LAG-3 protein or derivative is administered before, during or after administration of a chemotherapeutic agent, and the LAG-3 protein or derivative is administered to the subject at a dosage of >30 mg to <120 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0343] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject who has already been administered a chemotherapeutic agent, comprising administering to a subject in need of such prevention, treatment or amelioration a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, wherein the LAG-3 protein or derivative thereof is administered to the subject at a dosage of >30 mg to <200 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0344] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject who has already been administered a chemotherapeutic agent, comprising administering to a subject in need of such prevention, treatment or amelioration a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, wherein the LAG-3 protein or derivative thereof is administered to the subject at a dosage of >30 mg to <120 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0345] In yet a further embodiment, the present invention relates to a chemotherapeutic agent for use in the prevention, treatment, or amelioration of cancer in a subject, wherein the chemotherapeutic agent is administered to the subject at a dosage of >30 mg to <200 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321 before, during, or after administration of a LAG-3 protein or a derivative thereof capable of binding to an MHC class II molecule.

[0346] In yet a further embodiment, the present invention relates to a chemotherapeutic agent for use in the prevention, treatment, or amelioration of cancer in a subject, wherein the chemotherapeutic agent is administered to the subject at a dosage of >30 mg to <120 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321 before, during, or after administration of a LAG-3 protein or a derivative thereof capable of binding to an MHC class II molecule.

[0347] In another embodiment, the present invention relates to the use of a chemotherapeutic agent in the manufacture of a medicament for the prevention, treatment or amelioration of cancer in a subject, wherein the chemotherapeutic agent is administered to the subject at a dosage of >30 mg to <200 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321 before, during or after administration of a LAG-3 protein or a derivative thereof capable of binding to an MHC class II molecule.

[0348] In another embodiment, the present invention relates to the use of a chemotherapeutic agent in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject, wherein the chemotherapeutic agent is administered to the subject at a dosage of >30 mg to <120 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321 before, during, or after administration of a LAG-3 protein or a derivative thereof capable of binding to an MHC class II molecule.

[0349] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject who has already been administered LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, at a dose of >30 mg to <200 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321, comprising administering a chemotherapeutic agent to a subject in need of such prevention, treatment or amelioration.

[0350] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject who has already been administered LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, at a dose of >30 mg to <120 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321, comprising administering a chemotherapeutic agent to a subject in need of such prevention, treatment or amelioration.

[0351] Optionally, the LAG-3 protein or derivative thereof is administered to the subject at a dosage of between 40 mg and 200 mg of the molar equivalent of IMP321.

[0352] Optionally, the LAG-3 protein or derivative thereof is administered to the subject at a dosage of between 40 mg and 180 mg of the molar equivalent of IMP321.

[0353] Optionally, the LAG-3 protein or derivative thereof is administered to the subject at a dosage of between 40 mg and 150 mg of the molar equivalent of IMP321.

[0354] Optionally, the LAG-3 protein or derivative thereof is administered to the subject at a dosage of between 40 mg and 120 mg of the molar equivalent of IMP321.

[0355] Optionally, the LAG-3 protein or derivative thereof is administered to the subject at a dosage of between 50 mg and 200 mg of the molar equivalent of IMP321.

[0356] Optionally, the LAG-3 protein or derivative thereof is administered to the subject at a dosage of between 50 mg and 180 mg of the molar equivalent of IMP321.

[0357] Optionally, the LAG-3 protein or derivative thereof is administered to the subject at a dosage of between 50 mg and 150 mg of the molar equivalent of IMP321.

[0358] Optionally, the LAG-3 protein or derivative thereof is administered to the subject at a dosage of between 50 mg and 120 mg of the molar equivalent of IMP321.

[0359] Optionally, the LAG-3 protein or derivative thereof is administered to the subject at a dosage of the molar equivalent of 50-100 mg of IMP321.

[0360] Optionally, the LAG-3 protein or derivative thereof is administered to the subject at a dosage of 60-100 mg of IMP321, or the molar equivalent of 60-90 mg of IMP321.

[0361] Optionally, the LAG-3 protein or derivative thereof is administered to the subject at a dosage of the molar equivalent of 80-100 mg of IMP321.

[0362] Optionally, the LAG-3 protein or derivative thereof is administered to the subject at a dosage that is the molar equivalent of 90 mg of IMP321.

[0363] The dosage of LAG-3 protein or its derivative may be administered to a subject in two or more separate doses, each separate dose including a partial dose to provide a total dose in combination.For example, a 90mg dose of IMP321 may be administered in two separate doses of 45mg (e.g., by subcutaneous injection).The separate doses may be separated by up to 30 minutes, for example, up to 15 minutes.

[0364] Optionally, multiple doses of the LAG-3 protein or derivative thereof are administered to the subject.

[0365] Optionally, multiple doses of the chemotherapeutic agent are administered to the subject.

[0366] Optionally, the LAG-3 protein or derivative is administered to the subject on the same day as the chemotherapy agent.

[0367] Optionally, the LAG-3 protein or derivative is administered to the subject on days 1 and 15 of a 4-week cycle, and the chemotherapeutic agent is administered to the subject on days 1, 8, and 15 of the 4-week cycle, and optionally, the 4-week cycle is repeated for 4 to 8 cycles, preferably 6 cycles.

[0368] Optionally, the LAG-3 protein or derivative is administered to the subject on days 1 and 15 of a 4-week cycle, and the chemotherapeutic agent is administered to the subject on days 1, 8, and 15 of the 4-week cycle, and optionally the 4-week cycle is repeated for 4 to 13 cycles, preferably 6, 7, 8, 9, 10, 11, 12 or 13 cycles.

[0369] Optionally, the LAG-3 protein or derivative is administered to the subject one or more times in the absence of a chemotherapeutic agent, after which one or more doses of the LAG-3 protein or derivative are administered to the subject prior to, along with, or following one or more doses of the chemotherapeutic agent.

[0370] Optionally, the LAG-3 protein or derivative is administered to the subject in the absence of a chemotherapeutic agent on days 1 and 15 of a four-week cycle for up to nine four-week cycles.

[0371] Optionally, the LAG-3 protein or derivative is administered to the subject in the absence of a chemotherapeutic agent on days 1 and 15 of a four-week cycle for up to seven four-week cycles.

[0372] Optionally, the LAG-3 protein or derivative is administered to the subject over four to eight, preferably six, four-week cycles, in the absence of the chemotherapy agent following doses of the LAG-3 protein or derivative on days 1 and 15 of the four-week cycle, and doses of the chemotherapy agent on days 1, 8, and 15 of the four-week cycle.

[0373] Optionally, the LAG-3 protein or derivative is administered to the subject over 4 to 13, preferably 6, 7, 8, 9, 10, 11, 12 or 13, four-week cycles, in the absence of the chemotherapy agent following doses of the LAG-3 protein or derivative on days 1 and 15 of the four-week cycle, and doses of the chemotherapy agent on days 1, 8 and 15 of the four-week cycle.

[0374] Optionally, the chemotherapeutic agent is administered at a dosage in accordance with the approved prescribing information.

[0375] Optionally, the chemotherapeutic agent is a taxane.

[0376] Optionally, the chemotherapeutic agent is paclitaxel.

[0377] Optionally, the LAG-3 derivative is a LAG-3Ig fusion protein, IMP321.

[0378] Optionally, 80 mg / m 2 of paclitaxel will be administered intravenously to subjects on days 1, 8, and 15 of a 4-week cycle, followed by 90 mg of IMP321 administered subcutaneously on days 1 and 15 of a 4-week cycle.

[0379] Optionally, the LAG-3 derivative is a LAG-3Ig fusion protein, IMP321.

[0380] Optionally, the cancer is breast cancer.

[0381] Optionally, the cancer is metastatic breast cancer.

[0382] Optionally, the subject is a hormone receptor positive HER2-negative / low (HR + Patients with HER2-negative / low) metastatic breast cancer or metastatic triple-negative breast cancer (TNBC).

[0383] In a further embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent for use in the prevention, treatment, or amelioration of cancer in a subject, wherein the LAG-3 protein or derivative is administered to the subject on the same day as the chemotherapeutic agent.

[0384] In another embodiment, the present invention relates to the use of a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject, wherein the LAG-3 protein or derivative is administered to the subject on the same day as the chemotherapeutic agent.

[0385] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject, comprising administering to a subject in need of such prevention, treatment or amelioration a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent, wherein the LAG-3 protein or derivative is administered to the subject on the same day as the chemotherapeutic agent.

[0386] In yet a further embodiment, the present invention relates to a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, for use in the prevention, treatment, or amelioration of cancer in a subject, wherein the LAG-3 protein or derivative is administered before, during, or after administration of a chemotherapeutic agent, and wherein the LAG-3 protein or derivative is administered to the subject on the same day as the chemotherapeutic agent.

[0387] In another embodiment, the present invention relates to the use of a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject, wherein the LAG-3 protein or derivative is administered before, during, or after administration of a chemotherapeutic agent, and the LAG-3 protein or derivative is administered to the subject on the same day as the chemotherapeutic agent.

[0388] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject who has already been administered a chemotherapeutic agent, comprising administering to a subject in need of such prevention, treatment or amelioration a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, wherein the LAG-3 protein or derivative thereof is administered to the subject on the same day as the chemotherapeutic agent.

[0389] In yet a further embodiment, the present invention relates to a chemotherapeutic agent for use in the prevention, treatment, or amelioration of cancer in a subject, wherein the chemotherapeutic agent is administered to the subject before, during, or after administration of a LAG-3 protein or a derivative thereof capable of binding to an MHC class II molecule, and the chemotherapeutic agent is administered to the subject on the same day as the LAG-3 protein or derivative.

[0390] In another embodiment, the present invention relates to the use of a chemotherapeutic agent in the manufacture of a medicament for the prevention, treatment, or amelioration of cancer in a subject, wherein the chemotherapeutic agent is administered to the subject before, during, or after administration of a LAG-3 protein or a derivative thereof capable of binding to an MHC class II molecule, and the chemotherapeutic agent is administered to the subject on the same day as the LAG-3 protein or derivative.

[0391] In a further embodiment, the present invention provides a method for preventing, treating or ameliorating cancer in a subject who has already been administered a LAG-3 protein or a derivative thereof capable of binding to an MHC class II molecule, comprising administering a chemotherapeutic agent to a subject in need of such prevention, treatment or amelioration, wherein the chemotherapeutic agent is administered to the subject on the same day as the LAG-3 protein or derivative thereof.

[0392] Optionally, the LAG-3 protein or derivative is administered to the subject on days 1 and 15 of a 4-week cycle, and the chemotherapeutic agent is administered to the subject on days 1, 8, and 15 of the 4-week cycle, and optionally, the 4-week cycle is repeated for 4 to 8 cycles, preferably 6 cycles.

[0393] Optionally, the LAG-3 protein or derivative is administered to the subject on days 1 and 15 of a 4-week cycle, and the chemotherapeutic agent is administered to the subject on days 1, 8, and 15 of the 4-week cycle, and optionally the 4-week cycle is repeated for 4 to 13 cycles, preferably 6, 7, 8, 9, 10, 11, 12 or 13 cycles.

[0394] Optionally, the LAG-3 protein or derivative is administered at a dosage of 1 mg to 200 mg, 6 mg to 200 mg, 6 mg to <120 mg, 20 mg to 100 mg, 30 mg to 90 mg, about 30 mg, or about 90 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

[0395] Optionally, the chemotherapeutic agent is administered at a dosage in accordance with the approved prescribing information.

[0396] Optionally, the chemotherapeutic agent is a taxane.

[0397] Optionally, the chemotherapeutic agent is paclitaxel.

[0398] Optionally, the LAG-3 derivative is a LAG-3Ig fusion protein, IMP321.

[0399] Optionally, 80 mg / m 2 of paclitaxel will be administered intravenously to subjects on days 1, 8, and 15 of a 4-week cycle, followed by 90 mg of IMP321 administered subcutaneously on days 1 and 15 of a 4-week cycle.

[0400] Optionally, the cancer is breast cancer.

[0401] Optionally, the cancer is metastatic breast cancer.

[0402] Optionally, the subject is a hormone receptor positive HER2-negative / low (HR + Patients with HER2-negative / low) metastatic breast cancer or metastatic triple-negative breast cancer (TNBC).

[0403] <Combination treatment drug regimen> In one embodiment, the LAG-3 protein or derivative thereof is administered to the subject following administration of the chemotherapeutic agent, within about 12 to about 96 hours, about 12 to about 48 hours, or about 24 hours of administration of the chemotherapeutic agent.

[0404] According to another embodiment of the present invention, the combination treatment with chemo-immunotherapy comprises 6 cycles of 4 weeks. Patients are administered paclitaxel every week on days 1, 8 and 15, with adjuvant treatment with LAG-3 protein or its derivative on days 2 and 16 of each 4-week cycle. After completing the 6-cycle chemo-immunotherapy phase, responding or stable patients receive LAG-3 protein or its derivative every 4 weeks during the maintenance phase for a period of up to 12 injections (48 weeks).

[0405] In another embodiment, the chemo-immunotherapy combination treatment comprises 7 cycles of 4 weeks, or 8 cycles of 4 weeks, or 9 cycles of 4 weeks, or 10 cycles of 4 weeks, or 11 cycles of 4 weeks, or 12 cycles of 4 weeks (long-term combination treatment).

[0406] In other embodiments, after the chemoimmunotherapy phase, responding or stable patients receive the LAG-3 protein or derivative thereof about every week, or about every two weeks, or about every three weeks during the maintenance phase for up to about 48 weeks, or up to 96 weeks.

[0407] In another embodiment, the present invention relates to the use of LAG-3 protein or a derivative thereof as a maintenance therapy following cancer treatment according to an embodiment of the present invention. Suitably, during the maintenance therapy, the LAG-3 protein or a derivative thereof is administered about every week, or about every two weeks, or about every three weeks, or about every four weeks, for up to about 48 weeks, or for up to 96 weeks.

[0408] In a further embodiment, the present invention relates to a LAG-3 protein or a derivative thereof capable of binding to an MHC class II molecule for use in the prevention, treatment or amelioration of cancer in a subject, wherein the LAG-3 protein or derivative thereof is administered to the subject on the same day as a chemotherapeutic agent.

[0409] Optionally, the LAG-3 protein or derivative is administered to the subject on days 1 and 15 of a 4-week cycle, and the chemotherapeutic agent is administered to the subject on days 1, 8, and 15 of the 4-week cycle, and optionally, the 4-week cycle is repeated for 4 to 8 cycles, preferably 6 cycles.

[0410] Optionally, the LAG-3 protein or derivative is administered to the subject on days 1 and 15 of a 4-week cycle, and the chemotherapeutic agent is administered to the subject on days 1, 8, and 15 of the 4-week cycle, and optionally the 4-week cycle is repeated for 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13 cycles, preferably 6, 7, 8, 9, 10, 11, 12 or 13 cycles.

[0411] Optionally, the LAG-3 protein or derivative is administered to the subject on days 1 and 15 of a 4-week cycle, and the chemotherapeutic agent is administered to the subject on days 1, 8, and 15 of the 4-week cycle, optionally, the 4-week cycle is repeated for up to 13 cycles (52 weeks).

[0412] The length of treatment with the combination of the LAG-3 protein or derivative and the chemotherapeutic agent depends on how well the chemotherapeutic agent is tolerated by the patient. Preferably, at least six four-week cycles (24 weeks) of treatment with the LAG-3 protein or derivative and the chemotherapeutic agent are administered.

[0413] Optionally, 80 mg / m 2 of paclitaxel will be administered intravenously to subjects on days 1, 8, and 15 of a 4-week cycle, followed by 90 mg of IMP321 administered subcutaneously on days 1 and 15 of a 4-week cycle.

[0414] In a further embodiment, the present invention provides a LAG-3 protein or a derivative thereof capable of binding to an MHC class II molecule for use in the prevention, treatment, or amelioration of cancer in a subject, wherein the LAG-3 protein or derivative is administered to the subject one or more times in the absence of a chemotherapeutic agent after one or more doses of the LAG-3 protein or derivative have been administered to the subject before, with, or after one or more doses of a chemotherapeutic agent.

[0415] Optionally, the LAG-3 protein or derivative is administered to the subject in the absence of a chemotherapeutic agent on days 1 and 15 of a four-week cycle for up to nine four-week cycles.

[0416] Optionally, the LAG-3 protein or derivative is administered to the subject in the absence of a chemotherapeutic agent on days 1 and 15 of a four-week cycle for up to seven four-week cycles.

[0417] Optionally, the LAG-3 protein or derivative is administered to the subject over four to eight, preferably six, four-week cycles, in the absence of the chemotherapy agent following doses of the LAG-3 protein or derivative on days 1 and 15 of the four-week cycle, and doses of the chemotherapy agent on days 1, 8, and 15 of the four-week cycle.

[0418] Optionally, the LAG-3 protein or derivative is administered to the subject over 4 to 13, preferably 6, 7, 8, 9, 10, 11, 12 or 13, four-week cycles, in the absence of the chemotherapy agent following doses of the LAG-3 protein or derivative on days 1 and 15 of the four-week cycle, and doses of the chemotherapy agent on days 1, 8 and 15 of the four-week cycle.

[0419] Optionally, the combination of the chemo-immunotherapy phase (chemo-IO) and maintenance phase (IO) of treatment is for a total of up to 13 four-week cycles (52 weeks).

[0420] Optionally, the subject has not previously been treated with chemotherapy for metastatic disease.

[0421] Optionally, the hormone receptor positive HER2-negative / low (HR+ / HER2-negative / low) metastatic breast cancer patient is endocrine resistant.

[0422] Optionally, the patient with hormone receptor positive HER2-negative / low (HR+ / HER2-negative / low) metastatic breast cancer has been previously treated with a CDK4 / 6 inhibitor.

[0423] Optionally, the patient with hormone receptor positive HER2-negative / low (HR+ / HER2-negative / low) metastatic breast cancer is endocrine refractory and has previously been treated with a CDK4 / 6 inhibitor.

[0424] Optionally, patients with hormone receptor positive HER2-negative / low (HR+ / HER2-negative / low) metastatic breast cancer are endocrine refractory and have been previously treated with a CDK4 / 6 inhibitor and are aged 18-65 years.

[0425] Optionally, the metastatic triple-negative breast cancer (TNBC) patient is ineligible for anti-PD-1 or anti-PD-L1 based therapy (e.g., a combination of anti-PD-1 / PD-L1 therapy and chemotherapy).

[0426] Optionally, the metastatic triple-negative breast cancer (TNBC) patient is PD-L1 positive or PD-L1 negative.

[0427] Optionally, the metastatic triple-negative breast cancer (TNBC) patient is PD-L1 negative.

[0428] Optionally, the metastatic triple-negative breast cancer (TNBC) patient is between 18 and 65 years old.

[0429] In one particular embodiment, the present invention relates to a method of preventing, treating or ameliorating metastatic breast cancer in a subject, comprising administering to a subject in need of such prevention, treatment or amelioration a LAG-3 protein, or a derivative thereof capable of binding to an MHC class II molecule, and a chemotherapeutic agent, Here, the subject is a patient with hormone receptor positive HER2-negative / low (HR+ / HER2-negative / low) metastatic breast cancer or a patient with metastatic triple-negative breast cancer (TNBC), The LAG-3 protein or derivative thereof is administered to the subject on the same day as the chemotherapy agent; The LAG-3 protein or derivative thereof is administered to the subject at a dose that is the molar equivalent of about 6 mg to about 200 mg of the LAG-3 derivative LAG-Ig fusion protein IM321; The LAG-3 protein or derivative thereof and the chemotherapeutic agent are administered to the subject for at least six four-week cycles (24 weeks) of treatment.

[0430] Further patient subgroups In other embodiments, one or more additional patient subgroups are selected for treatment, including, for example, patients with higher or lower initial performance status, patients with extensive prior exposure to corticosteroids, and patients with a low BMI, e.g., <30 kg / m 2 Patients with

[0431] <Pharmaceutical Composition> The LAG-3 protein or derivative thereof and, if applicable, the chemotherapeutic agent are formulated together with a pharma- ceutically acceptable carrier, excipient, or diluent to provide a pharmaceutical composition. Typically, they are formulated as separate pharmaceutical compositions, but in the case of a fixed dose combination, the LAG-3 protein or derivative thereof and the chemotherapeutic agent are formulated together with a pharma- ceutically acceptable carrier, excipient, or diluent. The separate pharmaceutical compositions may be packaged together in the form of a kit of parts.

[0432] In general, the LAG-3 protein or derivative thereof and, where applicable, the chemotherapeutic agent may be administered by known means, in any suitable pharmaceutical composition, and by any suitable route.

[0433] Suitable pharmaceutical compositions may be prepared using conventional methods known to those skilled in the art of pharmaceutical formulation and described in the relevant texts and literature, for example, Remington: The Science and Practice of Pharmacy (Easton, Pa.: Mack Publishing Co., 1995).

[0434] For ease of administration and uniformity of dosage, it is particularly advantageous to formulate the composition of the present invention in unit dosage form.The term "unit dosage form" as used herein refers to a physically separate unit suitable as a unitary dosage for an individual to be treated.That is, the composition is formulated into individual dosage units, each containing a predetermined "unit dosage" amount of active agent calculated to produce a desired therapeutic effect in association with a required pharmaceutical carrier, excipient or diluent.The specification of the unit dosage form of the present invention depends on the unique characteristics of the active agent to be delivered.The dosage can further be determined with reference to the usual dose and mode of administration of the components.It should be noted that in some cases, two or more individual dosage units are combined to provide a therapeutically effective amount of active agent.

[0435] Formulations according to the invention for parenteral administration include sterile aqueous and non-aqueous solutions, suspensions, and emulsions. Aqueous solutions for injection contain the active agent in a water-soluble form. Examples of non-aqueous solvents or vehicles include fatty oils such as olive oil and corn oil, synthetic fatty acid esters such as ethyl oleate or triglycerides, low molecular weight alcohols such as propylene glycol, synthetic hydrophilic polymers such as polyethylene glycol, liposomes, and the like. Parenteral formulations may also contain adjuvants such as solubilizers, preservatives, wetting agents, emulsifiers, dispersants, and stabilizers, and aqueous suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, and dextran. Injectable preparations may be sterilized by incorporation of a sterilizing agent, filtration through a bacteria-retaining filter, irradiation, or heat. They may also be manufactured using a sterile injectable medium. The active agent may also be in a dry form, for example a lyophilized form, which can be rehydrated with a suitable vehicle immediately prior to administration by injection. EXAMPLES

[0436] Embodiments of the present invention will now be described, by way of example only, with reference to the accompanying drawings, in which:

[0437] Example 1 - HER2 - / HR + Active Immunotherapy Paclitaxel in Metastatic Breast Cancer (MBC) (AIPAC)] We conducted a multicenter, placebo-controlled, double-blind, 1:1 randomized phase IIb study in female patients with HER2-negative, hormone receptor-positive metastatic breast cancer. The study included two phases: Phase 1, an open-label safety run-in phase consisting of cohorts 1 and 2 to confirm the recommended Phase II dose (RPTD) of IMP321 in combination with paclitaxel; and · Phase 2, a placebo-controlled, double-blind, randomized phase comparing paclitaxel plus IMP321 to paclitaxel plus placebo in the RPTD.

[0438] experiment: IMP321 group: Paclitaxel + IMP321 at RPTD of 30 mg (114 patients): The chemoimmunotherapy phase consisted of six 4-week cycles. Patients received weekly paclitaxel on days 1, 8, and 15 with adjuvant treatment of the study drug (IMP321) on days 2 and 16 of each 4-week cycle. After completion of the six-cycle chemoimmunotherapy phase, responding or stable disease patients received the study drug (IMP321) every 4 weeks for an additional period of up to 12 injections during the maintenance phase.

[0439] Placebo group: Paclitaxel + placebo (112 patients): The chemoimmunotherapy phase consisted of six 4-week cycles. Patients received weekly paclitaxel on days 1, 8, and 15 with adjunctive treatment with study drug (placebo) on days 2 and 16 of each 4-week cycle.

[0440] After completion of the six-cycle chemoimmunotherapy phase, responding or stable disease patients received the study drug (placebo) for an additional period of up to 12 injections every 4 weeks during the maintenance phase.

[0441] result: Two hundred twenty-six patients were included in the full analysis set [IMP321 (n=114); placebo (n=112)]. Patients were endocrine refractory (84%), pretreated with a CDK4 / 6 inhibitor (44.2%), and had similar post-study treatment.

[0442] [Table 1] The database cutoff date was May 14, 2021 (73% of events) and the minimum follow-up was 22 months.

[0443] Exploratory univariate analyses (analysis arms defined before unblinding) showed that younger patients (<65 years), those with low baseline monocyte counts (<0.25 / nL), or breast cancer subtype luminal B had a significant and clinically meaningful improvement in median overall survival (OS) from treatment with IMP321. Multivariate analyses were performed using backward selection (p>0.15) from univariate Cox models.

[0444] [Table 2]

[0445] In multivariate predictive models, four patient subgroups (high neutrophil-to-lymphocyte ratio (NLR), no prior taxane treatment, low monocytes, and <5 years from diagnosis) were significant for improved OS.

[0446] Immune monitoring was also performed. Blood cell subsets (CD4, CD8, PBMC, monocytes) and Th1 biomarkers (CXCL-10) were measured centrally. Comparisons were performed using two-tailed Wilcoxon tests.

[0447] Fold changes in monocytes (5.81 vs. 2.29; p=0.025), PBMCs (2.00 vs. 1.41; p=0.041), T cells (2.28 vs. 1.48; p=0.086), and CXCL10 (2.78 vs. 1.56; p=0.06) on treatment were significantly higher compared to placebo and associated with improved OS. Post-baseline CD4 (median 896 / μl vs. 736 μl; p=0.038) and CD8 (median 377 / μl vs. 223 μl; p=0.005) T cell counts were significantly increased in patients with improved OS in the IMP321 arm versus placebo.

[0448] IMP321 added to paclitaxel induces significant effects on primary (monocyte) and secondary (CD4;CD8) target cells and related Th1 biomarkers that are significantly associated with improved OS.

[0449] Conclusion: The significant increase in OS in this specific patient subgroup, which represents a significant proportion of the overall patient population, is surprising and unexpected, as there have been no recent improvements in terms of treatment options for chemotherapy-eligible (i.e., post-endocrine therapy with or without CDK4 / 6 inhibitor treatment) HR+ / HER2- metastatic breast cancer patients. Furthermore, there are no active immunotherapies currently approved or in late-stage trials for this indication. This is due to the non-immunogenic nature of these types of tumors, which means that they are typically poorly or non-responsive to traditional immunotherapy options.

[0450] [Example 2 - Active Immunotherapy Paclitaxel-003 (AIPAC003) in HER2-negative / low metastatic breast cancer (MBC)] This example describes a planned randomized, double-blind, placebo-controlled Phase 3 study testing eftilagimod alpha (soluble LAG-3; IMP321), after open-label dose optimization, in patients with HER2-negative / low-metastatic breast cancer receiving paclitaxel.

[0451] Breast cancer (BC) is the most commonly diagnosed cancer worldwide, with an estimated 2.3 million new breast cancer cases reported in 2020, accounting for 11.7% of all cancer cases. In the United States, there were 253,465 estimated new cases in 2020 compared to 531,086 in Europe, and an estimated number of deaths was 42,617, i.e., 6.2% of all cancer deaths that year, compared to 141,765 in Europe.

[0452] Breast cancer is the leading cause of death worldwide in the female-only population with 685,000 deaths (the fifth leading cause of cancer deaths in both men and women). This equates to 1 in 4 cancer cases being the result of breast cancer in women, and 1 in 6 cancer deaths. With earlier detection and improved treatment, the 5-year net survival for BC has increased in recent years. Advanced / metastatic BC remains a virtually incurable disease, with a median OS of approximately 3 years and a 5-year survival rate of approximately 25%.

[0453] MBC patients are differentiated into four major molecular subtypes based on the degree of HER2 expression and HR status, as shown in Figure 2. + or HR - The aim of the study was to recruit patients with HER2-negative / low MBC. The majority of BC cases (68%) were HR + , i.e., estrogen receptor and / or progesterone receptor positive but HER2-negative / low. The 5-year survival rate is estimated at 30%.

[0454] HER2-high (previously HER2-pos) expression is defined as a 3+ score by immunohistochemistry (IHC) and / or a positive in situ hybridization (ISH) result (including 2+ in IHC and positive in ISH). All other patients are considered HER2-negative / low (Society, ACLast Revised: August 25, 2022. Breast Cancer HER2 Status). HER2-low BC represents a recently proposed classification of HER2 subtypes. Approximately half of all patients with breast cancer have tumors that are HER2-low. They have previously been classified as HER2-negative, with no effective treatment options with HER2-targeted drugs. HER2-low BC have a higher HR than patients with TNBC. + It is more common in patients with breast cancer. Metastatic triple-negative breast cancer (TNBC) is an aggressive disease with poor outcome and constitutes approximately 10-20% of all breast cancer cases. This type of breast cancer is characterized by the lack of expression of estrogen receptor (ER), progesterone receptor (PR) and HER2, as well as the presence of high histological grade and mitotic rate. The expected 5-year survival rate is estimated to be less than 15%.

[0455] This study is + and all patients with MBC who are either HER2-negative / low or TNBC and have previously received certain therapies.

[0456] <HR + HER2-negative / low> HR + The treatment landscape for HER2-negative / low MBC has undergone major changes in recent years. In the past, blockade of estrogen signaling with single-agent ET was the mainstay of first-line treatment. However, ET resistance eventually occurs in almost all patients, typically with disease progression 1 year after first-line single-agent ET. This treatment paradigm changed with the approval of CDK4 / 6 inhibitors. In 2015, palbociclib was the first CDK4 / 6 inhibitor approved. To date, multiple CDK4 / 6-based drugs have been approved by the FDA, EMA, and other health agencies in combination with different types of ET.

[0457] Early after introduction of CDK4 / 6 inhibitors, HR + HER2-negative / low MBC patients, in the first-line setting, typically receive ET alone or in combination with a CDK4 / 6 inhibitor, potentially followed by a later line of endocrine therapy (Figure 3). Chemotherapy was given when ET was not possible, e.g., in cases of acute life-threatening disease or endocrine resistance.

[0458] Today, the use of CDK4 / 6 inhibitors is widespread. Preferred regimens according to the NCCN / ESMO guidelines include CDK4 / 6 inhibitor and aromatase inhibitor (AI), fulvestrant ± CDK4 / 6 inhibitor, fulvestrant and nonsteroidal AI (category 1), and single-agent ET (category 2A). For second or subsequent lines, preferred regimens include fulvestrant + CDK4 / 6. For patients with PIK3 mutations, there is the option to receive a PI3 kinase inhibitor, as shown in Figure 3. Nevertheless, many patients become endocrine resistant at some point and require further treatment. Single-agent chemotherapy and especially taxanes are commonly used in this patient population (circles in Figure 3).

[0459] [Table 3] ORR: objective response rate; PFS: progression-free survival; OS: overall survival

[0460] The addition of CDK4 / 6 inhibitors dramatically increased PFS / OS for ET-based therapy, but different studies showed that chemotherapy-free overall survival decreased after CDK4 / 6 therapy. In the AIPAC study, OS in the paclitaxel + placebo arm decreased from 20.4 to 14.9 months (if previously treated with CDK4 / 6 compared to patients without previous CDK4 / 6 therapy). Prior CDK4 / 6 had a negative impact on PFS and OS, independent of other poor prognostic markers.

[0461] In general, patients currently undergoing chemotherapy have received many more lines of treatment and can be expected to be at a later stage (more metastatic sites, more involvement of the liver and other distant metastases) compared to 2015 and prior.

[0462] <tnbc> For TNBC patients presenting with metastatic disease, the choice of therapy depends primarily on PD-L1 expression, as presented in Figure 4. For patients with PD-L1 positive tumors (expressed in 38% of TNBC according to the Keynote-355 study), anti-PD-1 + chemotherapy is recommended.

[0463] The FDA and EMA approved atezolizumab in combination with nab-paclitaxel for the treatment of adult patients with unresectable locally advanced or metastatic PD-L1-positive TNBC in 2019 based on results from the phase 3 IMpassion130 trial (April 2018 cutoff). The indication was subsequently withdrawn in the United States in 2021 following results from the phase 3 IMpassion131 trial, in which a PD-L1 inhibitor plus paclitaxel did not demonstrate a statistically significant improvement in PFS over paclitaxel alone.

[0464] In 2021, both the EMA and FDA granted approval for pembrolizumab in combination with chemotherapy (paclitaxel, nab-paclitaxel, or carboplatin-gemcitabine) for patients with locally recurrent unresectable or metastatic TNBC whose tumors express PD-L1 (combined positive score (CPS) ≥ 10). Approval was based on KEYNOTE-355, a multicenter, double-blind, randomized, placebo-controlled study in patients with locally recurrent unresectable or metastatic TNBC who had not been previously treated with chemotherapy in the metastatic setting. Among patients with a CPS ≥ 10, median PFS was 9.7 months in the pembrolizumab chemotherapy group and 5.6 months in the placebo chemotherapy group (cutoff, December 11, 2019).

[0465] For patients who are not eligible for anti-PD-1 / PD-L1-containing therapy (approximately 62%), chemotherapy remains the primary systemic treatment, and international guidelines support the use of single-agent taxanes (with or without bevacizumab) or anthracyclines as first-line therapy, with a median OS of up to 18 months. Thus, patients with PD-L1-negative tumors or after failure of anti-PD-1 / PD-L1-based therapy have limited treatment options.

[0466] [Table 4]

[0467] Although improvements have been made in the adjuvant treatment of early stage breast cancer over the past few decades, approximately 30% of patients initially diagnosed with limited stage disease will eventually experience recurrence in the form of metastatic breast cancer (MBC). Additionally, 5-10% of patients may also be diagnosed with metastatic disease at the time of initial diagnosis. Despite major improvements for early and late stage BC patients with the introduction of novel therapies, no improvements have been made for patients who become eligible for chemotherapy with metastatic disease.

[0468] Today, HR + HER2-negative / low MBC patients receive chemotherapy later in the disease course and are more heavily pretreated at the time of chemotherapy initiation (Figure 3). + Patients with MBC acquire a more aggressive clinical behavior and eventually become refractory to any intervention. In the PALOMA-3 trial of palbociclib + fulvestrant, the duration of post-study chemotherapy was shortened from 6 to 5 months, which is a decrease of about 20% (Turner, et al., 2018. Overall Survival with Palbociclib and Fulvestrant in Advanced Breast Cancer. N Engl J Med 379:1926-1936). In multivariate analysis in AIPAC, previous CDK4 / 6 treatment was an independent poor prognostic marker, with a HR of 1.374 for OS and 1.65 for PFS. The median OS observed in AIPAC in patients receiving paclitaxel + placebo is clearly lower in patients pretreated with CDK4 / 6 (median 14.9 months) compared to historical data with a median OS of about 24 months for patients receiving single-agent chemotherapy for MBC.

[0469] For TNBC, novel therapies are urgently needed given the suboptimal outcomes of chemotherapy alone and the lack of treatment, especially in patients who are ineligible for anti-PD-1 / PD-L1 therapy (see Figure 4). Furthermore, no active immuno-oncology (IO) treatments have been approved for this patient population.

[0470] In conclusion, HR who have exhausted endocrine-based treatment options + There remains a high unmet medical need for HER2-negative / low MBC patients or TNBC patients who are ineligible for anti-PD-L1 / PD-1-based therapy and therefore candidates for chemotherapy such as weekly paclitaxel, with a median OS ranging from 12 to 18 months.

[0471] <Study design> The study consists of an open-label, dose-optimized run-in component followed by a double-blind, randomized, placebo-controlled Phase 3 component, as shown in Figure 5.

[0472] The dose optimization run-in phase includes two parts: a safety run-in followed by a randomized dose optimization run-in.

[0473] Dose-optimized implementation components In the initial open-label run-in component, the optimal biological dose (OBD) of efti in combination with weekly paclitaxel will be determined based on the following parameters: · Safety 1 and 2: % of patients with DLT and % of patients with adverse reactions leading to permanent discontinuation of efti within the first 4 cycles (excluding hypersensitivity to paclitaxel with C1D1). Tolerability: % of patients with any injection site reaction of any grade lasting >3 days and % of patients with any injection site reaction of any duration and ≥Grade 3 severity. · Efficacy: % of patients with BOR of PR / CR by RECIST 1.1. · PDM1 and 2: % of patients with a 1.4-fold increase in each PDM (PDM1-peripheral CD8+ T cells and PDM2-absolute lymphocyte count) compared to baseline within the first 4 cycles.

[0474] The OBD will be determined based on the data for both dose levels once at least 29 evaluable patients per dose cohort have been randomized. The decision algorithm is performed at three different levels, as outlined in Figure 6.

[0475] <Efficacy: Best Overall Response (BOR)> Overall response rate (ORR) was positively correlated with improvement in overall survival in the AIPAC trial, as shown below. Patients with a BOR of PR or CR according to RECIST 1.1 had a higher likelihood of overall survival > median. The Kaplan–Meier plots were significantly different (see Figure 7). The median decreased significantly (p < 0.001) from 27.5 [95% CI 20.6; 33.0] to 12.9 (95% CI 10.7–16.5) for patients without PR / CR as BOR. The HR was 0.5 [95% CI 0.37; 0.69], with a p-value < 0.001 in support of BOR of PR / CR, making ORR a good surrogate endpoint for the efficacy of this combination (OS, which is primary in the phase 3 portion), and thus selected as an efficacy marker.

[0476] <PDM1 - CD8 + Absolute number of T cells> In the AIPAC trial, in patients (n = 75) in whom fresh whole blood was directly analyzed ex vivo using FACS, a sustained statistically significant increase in circulating CD8 + T cells was observed in the efti group but not in the placebo group and was found to correlate with OS (Figure 8). This effect was seen only in the efti group, in samples taken at the time of treatment. Notably, the samples were taken 2 weeks after the previous efti dose (note that efti disappears from the bloodstream 3–4 days after injection), and thus, immediately prior to the next efti injection, the residual of efti in the trial was minimal. On the other hand, the pre-treatment levels of CD8+ T cells did not correlate with OS in either treatment group. Efti increases secondary target cells (CD8+ T cells) in peripheral blood, leading to improvement in OS. Based on the above reasons, the increase in circulating CD8+ T cells is a good surrogate for efti activity, correlates positively with overall survival, can be measured in a standardized way, and thus was selected as PDM1.

[0477] <PDM2 - Absolute lymphocyte count (ALC)> Absolute lymphocyte counts showed an early (after 4 weeks) and sustainable increase (≥0.2 / nl) within the EFTI group (green line, Figure 9). Increased ALC is associated with improved survival in patients treated with EFTI but not in placebo-treated patients (Figure 9). Just under half of the patients in AIPAC had an absolute increase of ≥0.2 / nl, which leaves room for improvement in the 90 mg cohort. Because this increase is observed early, it can be used in the future as a potential early-trial biomarker for treatment continuation. Based on the above reasons, absolute increase in lymphocytes is a good surrogate for EFTI activity, positively correlates with overall survival, and can be measured in a standardized manner, and was therefore selected as PDM2.

[0478] Phase 3 Components In the phase 3 component of the study, approximately 771 HER2-negative / low MBC patients will be randomized 2:1 to Arm A (active arm): paclitaxel + efti at OBD and Arm B (control arm): paclitaxel + placebo. Thus, there is a 66% probability that a patient will be randomized to the active treatment arm, and all patients will receive weekly paclitaxel as standard of care. In this study, the use of placebo in conjunction with double blinding greatly improves the quality of the data and its interpretation.

[0479] Patients were classified into (1) MBC type (HR + Against HR - ), (2) stratified by prior CDK4 / 6 (yes vs. no) and age (<65 years and ≥65 years). The study is event-driven and will be considered completed after recording the required number of events.

[0480] Dose-optimized induction and phase 3 schedule Each patient will undergo a screening period of up to 3 weeks. Treatment for the dose-optimized induction and phase 3 components of the study will consist of a chemo-immunotherapy (chemo-IO) phase followed by an immunotherapy (IO) phase (Figure 10). The chemo-IO phase will consist of 6 cycles of 4 weeks (28 days) each. Six cycles of paclitaxel will be planned. If paclitaxel is well tolerated, patients may continue beyond 6 cycles at the investigator's discretion based on the patient's individual tolerability. If paclitaxel needs to be stopped due to toxicity prior to the completion of 6 cycles, patients will be allowed to transition to efti / placebo alone (IO phase) if 4 cycles with paclitaxel have been completed.

[0481] During each chemo-IO cycle, patients received 80 mg / m on days 1, 8, and 15 in both arms of a 28-day (4-week) cycle. 2 Paclitaxel is administered intravenously, followed by 30 mg or 90 mg of efti in the dose-optimized induction, and placebo or efti OBD in the phase 3 component, subcutaneously on days 1 and 15 of a 28-day (4-week) cycle. The IO phase is planned to start in cycle 7 with efti or placebo (the latter only applies in arm B of phase 3) administered subcutaneously on days 1 and 15 of each cycle. The IO phase can start earlier or later, depending on each individual patient's paclitaxel resistance. Up to 13 cycles (approximately 12 months) of treatment are planned. A schematic diagram of the screening, treatment and follow-up periods is shown in Figure 10.

[0482] Patients will continue treatment until disease progression, unacceptable toxicity, completion of study treatment, or discontinuation for any other reason. The treatment period will end with an end-of-treatment (EOT) visit 1-3 weeks after the last study drug administration. Follow-up for progression (until disease progression in case patients no longer shows disease progression at EOT) and survival (until death in all patients) will occur until study termination, withdrawal of consent, or loss to follow-up, whichever occurs first.

[0483] Radiological assessments will be performed at 8-week intervals until week 32 and then every 12 weeks. They will be assessed according to Response Evaluation Criteria in Solid Tumors (RECIST) version 1.1 at the clinical site (investigator read) and treatment decisions will be based on the investigator read.

[0484] Appropriateness of Eftilagimod Alpha (Efti) at a dose of 30 mg The desired PD effects in vitro and ex vivo, i.e., activation of primary and secondary immune cells as exemplified by induction of cytokines, are observed starting at efti concentrations of approximately 1-10 ng / mL. In preclinical tumor efficacy models, the combination of efti with chemotherapy or PD-1 antagonists showed synergistic effects at dose levels corresponding to estimated plasma Cavg24h in the range of 2-5 ng / mL. Thus, the in vitro effective concentration range also appears to be valid in proof-of-concept animal studies. Available clinical data indicate that at a 30 mg subcutaneous dose in cancer patients, a comparable Cavg24h of 3.3 ng / mL is achieved. At a 6 mg dose, a lower Cavg24h of 0.8 ng / mL was observed.

[0485] Clinically administered Efti doses of 6 and 30 mg resulted in a plasma Cmax of ≥ 1 ng / mL in most patients, reflecting the minimum concentration for the desired efti PD effect. However, systemic efti levels > 1 ng / mL and sustained for ≥ 24 hours were observed in approximately 90% of patients receiving efti 30 mg, compared with only 20% receiving 6 mg.

[0486] PD markers related to T cell immune responses, such as CD4+ and CD8+ T cell counts and activation status, as well as related T cell cytokines such as IFN-γ and CXCL10, showed significant changes in patients after administration of efti every 2 weeks, while the percentage of patients with at least one of the aforementioned markers upregulated (defined as 1.4-fold compared to baseline) increased from approximately 50% of patients at 1 mg to approximately 90% at 30 mg. 30 mg is considered to be a safe and tolerable dose. The 30 mg dose has been widely tested in combination with pembrolizumab or weekly paclitaxel in NSCLC, HNSCC, melanoma and HR+MBC, showing promising efficacy compared to historical controls.

[0487] Appropriateness of Eftilagimod Alpha (Efti) at a dose of 90 mg It is recognized that 30 mg efti is the active dose, but based on the excellent safety profile and limited effect on PFS, investigation of higher doses in combination with weekly paclitaxel is considered. The 90 mg efti dose was selected based on PK simulations performed for three different dose levels (30 mg, 60 mg and 90 mg) and various parameters (e.g., Cmax, AUC0-48h and Cavg24h) using available popPK models. The AUC0-48h and Cavg24h 75% percentiles for 30 mg do not overlap with the 25% percentiles for 90 mg, ensuring that at least 50% of patients are expected to have higher AUC0-48h and Cavg24h in the 90 mg group compared to the 30 mg group. The distribution of simulated concentration vs. time profiles at 60 mg and 90 mg compared to 30 mg led to the same conclusion. The median Cmax is expected to be in the range of approximately 15 ng / mL for the 90 mg dose. In the available PK database, there were five patients with a Cmax >10 ng / mL at dose 1, and there were no safety concerns identified in these patients, i.e., no grade ≥ 3 adverse events related to efti, and three of the five patients reported a total of eight treatment-emergent adverse events with onset dates after the first dose and before the second dose, all of which were assessed by the investigator as related to study treatment. These adverse reactions (ARs) were mild (62.5%) or moderate (37.5%) in severity, and all but one recovered completely and had no effect on study treatment due to these events. In nonclinical toxicity studies, the NOAEL dose was defined as 15 mg / kg, and based on a patient weighing 60 kg, the 90 mg dose corresponds to 15 mg / kg.

[0488] The fact that the 90 mg dose is administered in two separate injections of 45 mg (1.8 mL) each further reduces any potential tolerability risk. This rationale is based on volume limitations, i.e., subcutaneous injection volumes exceeding 2 mL are associated with various problems including injection pain, adverse events at the injection site, and injection site leakage (i.e., backflow of the injection solution). Considering that the two injections are administered at a maximum interval of 15 minutes, no impact on the PK profile is expected compared to a single injection of 30 mg (1.2 mL).

[0489] Overall, the uniform administration of efti for dose optimization appears to be appropriate for immunostimulants with a wide therapeutic index.

[0490] <Validity of the Route and Frequency of Eftilagimod Alpha (Efti)> The route of administration of efti is subcutaneous. Primary target cells, i.e., dendritic cells, are abundant in the subcutaneous tissue. The choice of the route of administration is based on the principle that efti as an agonist does not reach a rapid peak of exposure that occurs after intravenous (i.v.) administration with the risk of pro-inflammatory systemic events, but is delivered to its cell target at low concentrations over a long period (at least 24 hours).

[0491] The rationale for the dosing frequency is that efti as an agonist continuously stimulates immunomodulatory factors over several months. This stable activation of effector T cells is desirable without affecting tolerability. It is known that the secondary T cell response to antigenic peptides shows a rapid kinetics with peak CD8+ T cell proliferation on day 7 in vitro, and that activated CD8+ T cells need to rest for an additional week so that they can respond to new antigenic stimuli again. Overall, a dosing schedule every two weeks is considered the best and most potent dosing schedule for patients with progressive or metastatic cancer who require effective immunostimulation without inducing concerns about safety or local tolerability.

[0492] In combination with anti-PD-1 and anti-PD-L1 therapy, efti is given on the same day without additional safety observations (IB ed 9.0). Based on this experience, we adapted the schedule applied in AIPAC to limit the number of days at the center for each patient (3 vs. 5 per 28-day schedule), administering efti on the same day as paclitaxel, ≥ 30 min after the end of the paclitaxel infusion.

[0493] <Appropriateness of Paclitaxel Dose> Weekly administration of paclitaxel has been evaluated to increase dose density and improve tolerability. In large randomized trials, weekly paclitaxel has been shown to be superior in terms of safety and efficacy compared to application every 3 weeks (D'Amico,et al.,2021.Standard of Care in Hormone Receptor-Positive Metastatic Breast Cancer:Can We Improve the Current Regimens or Develop Better Selection Tools? American Society of Clinical Oncology 18:331-334;Huard,et al.,1997.Characterization of the major histocompatibility complex class II binding site on LAG-3 protein.Proc Natl Acad Sci USA 94:5744-5749). Paclitaxel 80 or 90 mg / m 2 data on combination with efti is already available from AIPAC and shows that this dose is 90 mg / m 2 80 mg / m 2 The doses used in AIPAC-003 are: Paclitaxel 80 mg / m given on days 1, 8, and 15 of a 28-day cycle. 2 Monotherapy is a widely used regimen for the treatment of metastatic breast cancer (Decker,et al.,2017.A randomized phase II study of paclitaxel alone versus paclitaxel plus sorafenib in second-and third-line treatment of patients with HER2-negative metastatic breast cancer(PASO).BMC Cancer 17:499;Hernandez-Aya and Ma.2016.Chemotherapy principles of managing stage IV breast cancer in the United States.Chinese Clinical Oncology 5:42;Keane,M.2021.PACLitaxel 80mg / m2 Day 1,8 and 15 Monotherapy-28 Day.1 ed.NCCP,HSE homepage,National Cancer Control Programme;2022.Regimen Reference Order-BRST-PACLitaxel Metastatic every 7 days.CancerCare Manitoba,CancerCare Manitoba webpage.2).

[0494] If a patient wishes to participate in a study, they must meet the following criteria (among others): 1.Metastatic HR + (estrogen receptor positive and / or progesterone receptor positive) or hormone receptor negative (HR-), and HER2-negative / low-grade breast cancer was histologically proven by biopsy on the last available tumor tissue (primary tumor and / or metastasis; metastatic was preferred). Note: Estrogen and / or progesterone receptor positivity is defined as ≥1%, and HER2 receptor negativity is defined in line with ASCO / CAP guidelines (Burstein, et al. 2021. Endocrine Treatment and Targeted Therapy for Hormone Receptor-Positive, Human Epidermal Growth Factor Receptor 2-Negative Metastatic Breast Cancer: ASCO Guideline Update. J Clin Oncol: Jco2101392; Wolff, et al. 2013. Recommendations for human epidermal growth factor receptor 2 testing in breast cancer: American Society of Clinical Oncology / College of American Pathologists clinical practice guideline update. J Clin Oncol 31:3997-4013.). This HER2-low category includes those with IHC scores of 1+ and 2+ (measured by their ISH test) without amplification. Estrogen-, progesterone-, and HER2 receptor testing will be performed according to local guidelines (Wolff, AC 2018. HER2 Testing in Breast Cancer: American Society of Clinical Oncology / College of American Pathologists Clinical Practice Guideline Focused Update Summary. Journal of Oncology Practice: 14(17):437-441) (e.g., ASCO / CAP guidelines). 2.HR + Patients with MBC who have progressed after ≥ 1 line of endocrine-based therapy and are indicated to receive chemotherapy for metastatic disease. a. Primary endocrine resistance: recurrence / relapse ≤2 years after initiation of adjuvant endocrine therapy for early stage breast cancer or progression of first-line endocrine therapy within 6 months for metastatic breast cancer. b. Secondary endocrine resistance: recurrence / relapse >2 years after initiation of adjuvant endocrine therapy, recurrence / relapse <12 months after completion of adjuvant endocrine therapy, or progression after >6 months of endocrine therapy for metastatic breast cancer. Note: Previous targeted therapy (e.g., CDK4 / 6 or mTOR inhibitors) is acceptable. Patients with known PI3K mutations may have received a PI3K inhibitor prior to inclusion. Patients with BRCA mutations may have received a PARP inhibitor prior to inclusion. Apart from conventional cytotoxic chemotherapy, any treatment given in combination with endocrine therapy as first-line treatment for MBC is acceptable. 3. Patients with TNBC who have been shown to receive paclitaxel chemotherapy without anti-PD-1 / PD-L1 therapy in the first-line setting for metastatic disease. Note: Prior anti-PD-1 / PD-L1 therapy in the adjuvant setting is permitted. Patients with BRCA mutations may have received a PARP inhibitor prior to enrollment. 4. Dose optimization induction: Women aged 18 years or older. Phase 3: Women or men aged 18 years or older. 5. All patients of childbearing potential must have a negative high-sensitivity pregnancy test at screening and agree to use highly effective methods of contraception in accordance with EU Clinical Trial Facilitation Group guidance from the start of the study until at least 6 months after the last dose of study drug. Partners of patients of childbearing potential must also apply contraception. Patients who are any of the following: a. Postmenopausal (≥60 years, or <60 years and amenorrhea for 12 months in the absence of chemotherapy, tamoxifen, toremifene, or ovarian suppression with follicle-stimulating hormone (FSH) >40 U / L and estradiol <30 ng / L; or FSH and estradiol in the postmenopausal range if taking tamoxifen or toremifene and age <60 years), permanently sterilized (e.g., bilateral tubal occlusion, hysterectomy). b. Infertility is not considered to be a possibility for conception. 6. Dose-optimized induction only: evidence of measurable disease as defined by RECIST 1.1. Note: Patients with non-measurable disease as defined by RECIST 1.1 may be enrolled in the Phase 3 component. 7.Inspection Standards: a. Total white blood cell count ≧3×10 9 / L b. Platelet count ≧100×10 9 / L C. Hemoglobin ≥ 9 g / dL or 5.58 mmol / L Absolute neutrophil count (ANC) ≥ 1.5 x 10 9 / L e. CKD-EPI serum creatinine clearance >30mL / min f. Total bilirubin ≦20 μmol / L, except for familial cholestasis (Gilbert's disease). g. In the presence of liver metastases, serum ASAT and ALAT are ≤3x ULN or ≤5x ULN

[0495] Patients should be excluded from the study for any of the following reasons (among others): 1. Prior chemotherapy for metastatic breast adenocarcinoma. Note: Only regimens given in the metastatic setting and containing traditional cytotoxic chemotherapy agents are considered grounds for exclusion. Prior treatment with targeted agents of any type may be acceptable if all other applicable criteria are met. 2.HR + Patients with MBC who have received <1st line of ET-based therapy in the metastatic setting. 3.HR + Patients with MBC who are not primary or secondary resistant to ET-based therapy and who would be candidates for ET-based therapy according to applicable treatment guidelines. 4. TNBC patients who are candidates for PD-1 / PD-L1 therapy in combination with chemotherapy.

[0496] <Test Treatment> (Identity of investigational medicinal products (test drugs)) Eftilagimod alpha The efti drug product is a single-use, preservative-free, sterile efti solution for subcutaneous injection at a concentration of 25±1.2 mg / mL. The formulation is filled into 2 mL glass vials with an extractable fill volume of ≥1.2 mL and stored at 2°C-8°C, protected from light.

[0497] (Placebo vs. Eftilagimod alpha) The placebo was matched to the EFTI drug product for appearance and injectable properties. Placebo was filled into 2 mL glass vials with an extractable fill volume of ≥ 1.2 mL and stored at 2-8 °C, protected from light.

[0498] <Dosage regimen> Eftilagimod alpha Efti will be administered at doses of 30mg and 90mg during the dose-optimized induction component. Repeated subcutaneous doses of efti will be administered on days 1 and 15 of a 4-week cycle during the chemo-IO and IO components. Efti will be administered at least 30 minutes after the paclitaxel infusion is completed. The maximum number of efti doses is 26.

[0499] Efti (at OBD) or placebo will be administered during the Phase 3 component following the same regimen as the dose-optimized lead-in component.

[0500] The route of administration for efti is subcutaneous injection. For the 30 mg dose (1.2 mL), a single anatomical site is used on the anterior thigh. The location of the injection site should alternate between thighs with each injection (e.g., if the first injection on C1D1 is given in the left thigh, the next injection on C1D15 is given in the right thigh, etc.).

[0501] The 90 mg dose will be administered as two separate injections of 45 mg (1.8 mL) (timed a maximum of 15 minutes apart) as follows: a single anatomical site will be used on the anterior of one thigh and a single anatomical site will be used on the anterior of the other thigh.

[0502] The injection should be given slowly to avoid discomfort at the injection site.

[0503] (Paclitaxel) Paclitaxel was administered at 80 mg / m on days 1, 8, and 15 of a 4-week cycle. 2 (maximum 160 mg for patients with a body surface area ≥ 2 m2) is administered as a 1-hour intravenous (iv) infusion.

[0504] Methods of administration and supportive care follow local clinical practice, with the exception of corticosteroid premedication, for which the following restrictions apply: On days 1 and 8 of the first treatment cycle (i.e., before the first two paclitaxel infusions), 10 mg of dexamethasone is administered intravenously only once, 30 minutes before the paclitaxel infusion. Oral corticosteroid premedication or corticosteroid premedication started earlier than specified is not permitted. If the first two intravenous infusions of paclitaxel (one cycle) are well tolerated, no further premedication with corticosteroids should be administered. In patients who develop a reaction to paclitaxel, 10 mg of dexamethasone (or an equivalent dose of other systemic corticosteroids) is the maximum dose for premedication for all following paclitaxel administration.

[0505] There are no limitations to other means of premedicating paclitaxel for any of the treatment cycles, including but not limited to antihistamines.

[0506] Six cycles of paclitaxel are planned. If paclitaxel is well tolerated, patients may be continued beyond six cycles at the investigator's discretion based on the patient's individual tolerability.

[0507] If paclitaxel needs to be stopped due to toxicity prior to completion of 6 cycles, patients will be allowed to transition to efti / placebo alone (IO phase) upon completion of 4 cycles with paclitaxel.

[0508] Patients will continue treatment until disease progression, unacceptable toxicity, completion of study treatment, death, or discontinuation for any other reason.

[0509] Commercially available paclitaxel products will be used. Formulations other than the conventional formulations, such as paclitaxel drugs formulated as albumin-bound nanoparticles (nab-paclitaxel), are strictly not permitted in the study.

[0510] <Test evaluation> Efficacy evaluation (Radiological scans and color digital photographs) Computed tomography (CT) scans with contrast of the chest, abdomen, and pelvis are required for each patient at each time point. Tumor imaging is highly preferably obtained by CT with contrast enhancement. For the abdomen and pelvis, contrast-enhanced magnetic resonance imaging (MRI) can be used if CT with iodinated contrast is contraindicated or mandated by local practice, and for the chest, non-contrast CT of the chest is recommended to evaluate the lung parenchyma.

[0511] If brain imaging is performed to document stability of existing metastases or to rule out / confirm suspicious new brain metastases during the course of the study, MRI is the highly preferred modality (not mandatory for patients without evidence of brain disease). If brain MRI is contraindicated for any reason, a head CT including the brain should be performed instead. To optimize reproducibility of assessment of existing and new tumor burden and improve accuracy of imaging-based assessment of response or progression, the same imaging technique in terms of modality, ideally the same scanner, and use of contrast agent should be used for patients throughout the study (see Imaging Acquisition Guidelines).

[0512] MRI scans of the head and color digital photography of skin lesions will be performed as clinically indicated.

[0513] MRI examinations should be performed using a 1.5T or 3T MRI system.

[0514] The machine used (CT or MRI) is described in the imaging manual. The process for image acquisition and transmission to a central imaging vendor can be found in the Imaging Acquisition Guidelines.

[0515] Lesions identified at follow-up in anatomical locations not scanned at screening are considered new lesions and treated accordingly. An example of this would be if a patient had visceral disease at screening and required a CT or MRI scan of the brain that would reveal metastases during the study. A patient's brain metastases would be considered new lesions, even if brain imaging was not performed at screening.

[0516] Patients with symptomatic brain and / or leptomeningeal metastases will not be permitted to participate in the study. For all other patients, study-specific radiological scans of the brain are not foreseen, except for patients with abnormalities during the neurological screening evaluation, at the discretion of the investigator.

[0517] If skin lesions are present at screening, they will be evaluated at the same intervals as above. Color digital photography will be performed if any skin lesions are present between all time points scheduled for radiological evaluation.

[0518] All treatment decisions are based on investigator review. Images will be collected for retrospective blinded independent central review.

[0519] <Imaging> Imaging is performed to determine tumor burden according to the following schedule. Screening (Induction and Phase 3): ≤21 days prior to treatment initiation, historical images (obtained within a 6-week window prior to initiation of study treatment) may be used to assess patient eligibility if they follow the imaging protocol described in the Imaging Acquisition Guidelines. For patients in the induction and Phase 3 components, radiological assessments performed according to RECIST 1.1 should be performed before receiving any treatment during screening, and every 8 weeks until Week 32, and every 12 weeks thereafter. It may be performed ±3 days prior to a specific radiological assessment until Week 32. The imaging schedule is not affected by treatment and begins from the date of randomization.

[0520] Actions with the study treatment / study medication (e.g., treatment delays) will not affect the imaging schedule. The imaging schedule will remain as determined from randomization.

[0521] Clinical Lesions: Clinical lesions detected by physical examination are considered measurable only if they are superficial and ≥10 mm in diameter as measured with a caliper (e.g., skin nodules). If skin lesions are present at screening, documentation of these visible lesions (e.g., indexing tumor areas and / or new skin lesions) by color photography including a centimeter ruler to estimate the size of the lesion is recommended. All photographs will maintain patient anonymity and will be labeled with the patient's study identifier and the date of the photograph.

[0522] If skin lesions can be evaluated by both clinical examination and imaging, imaging evaluation should be performed as it provides a more objective assessment.

[0523] Superficial clinical lesions (e.g., skin nodules) must be measured in at least one dimension (the longest diameter in the plane of measurement is recorded) with a minimum size of the longest diameter ≥ 10 mm as imaged on a color photographic scale according to RECIST 1.1.

[0524] If a patient clinically presents with skin lesions, the investigator's facility will perform color digital photography of all skin lesions, using a ruler held flush against the skin next to the longest diameter of the lesion, to indicate the size of the lesion at all time points scheduled for radiology scans where the lesion is present. Once a lesion is documented, the area of ​​interest should be documented at every subsequent time point for the duration of the study.

[0525] Lesions identified at follow-up in an anatomical location not scanned at baseline are considered new lesions and indicate disease progression. An example of this is when a patient has visceral disease at baseline and requires a CT or MRI scan of the brain that reveals metastases during the study. The patient's brain metastases are considered evidence of PD even if brain imaging was not performed at baseline. In addition, patients who require a bone scan undergo a nuclear medicine-based bone scan (e.g., 99mTc-MDP bone scan). Every single new foci of radiotracer uptake is confirmed by either CT, MRI, or biopsy to demonstrate progression. The presence of two or more new foci in the absence of a benign cause (e.g., fracture or trauma) is considered evidence of PD even if there was no bone imaging at baseline.

[0526] (Progression-free survival) PFS is defined as the number of days between the date of treatment assignment (run-in component) or randomization (phase 3 component) and the time to documented disease progression or death from any cause, as assessed by investigator assessment per RECIST 1.1. The date of disease progression or censoring for PFS is determined according to the conventions listed below. These conventions are based on the December 2018 FDA Guidance for Industry, "Clinical Study Endpoints for the Approval of Cancer Drugs and Biologics" (https: / / www.fda.gov / media / 71195 / download) and the April 2015 FDA Guidance for the Industry, "Clinical Trial Endpoints for the Approval of Non-Small Cell Lung Cancer Drugs and Biologics" (https: / / www.fda.gov / media / 116860 / download).

[0527] [Table 5]

[0528] (overall survival) Overall survival (OS) is defined as the time between the date of treatment assignment (run-in component) or from randomization to death from any cause.

[0529] Patients lost to follow-up and patients alive at the date of data cut-off will be censored at the date the patient was last known to be alive or the date of data cut-off, whichever occurred first.

[0530] Overall survival follow-up visits should occur every 12 weeks after enrollment in the study for the first 2 years, and every 24 (+ / - 4) weeks thereafter. Visits can be conducted by telephone. In addition, any next lines of anticancer therapy will be recorded. If necessary, patients may be contacted occasionally outside of this FU window. OS-FU will be performed until study completion, death, withdrawal of consent, or loss to follow-up, whichever occurs first.

[0531] (Tumor response) Only patients with measurable disease at screening will be included in the run-in component of the study. Patients with non-measurable disease may be enrolled in the Phase 3 component of the study.

[0532] If two or more measurable lesions are present at baseline, all lesions up to a total of five lesions (and a maximum of two lesions per organ), representing all involved organs, should be identified as target lesions and evaluated at baseline and at specific time points throughout the study. Tumor response to target lesions will be assessed according to RECIST 1.1: Complete response (CR): Disappearance of all target lesions Partial Response (PR): At least a 30% reduction from baseline in the sum of the longest diameters of target lesions (longest axis of non-nodular lesions and short axis of nodular areas) Progressive Disease (PD): At least a 20% increase in the sum of the longest diameters of the target lesions (longest diameter of non-nodular lesions, short axis of nodular areas) based on the smallest sum during the study Stable Disease (SD): Minor changes not meeting the above criteria

[0533] All other lesions (or disease sites) should be identified as non-target lesions and will also be evaluated at baseline and at specific time points throughout the study. Tumor response for the non-target lesion group will be evaluated according to RECIST 1.1: CR: Disappearance of all non-target lesions and normalization of CA15-3 tumor marker levels. All lymph nodes must be non-pathological in size (<10mm short axis). · Non-CR / Non-PD: persistence of one or more non-target lesions and / or maintenance of CA15-3 tumor marker levels above normal limits. ·PD: Heterogeneous progression of pre-existing non-target lesions.

[0534] At each protocol point, the overall response is determined as follows:

[0535] [Table 6] < / tnbc>

Claims

1. 1. A LAG-3 protein or a derivative thereof capable of binding to an MHC class II molecule for use in preventing, treating, or ameliorating metastatic breast cancer in a subject with one or more of: low monocyte count; luminal B breast cancer; age less than about 85 years; having previously been treated with a CDK4 / 6 inhibitor; not having previously been treated with taxane chemotherapy; having an elevated neutrophil-to-lymphocyte ratio; and being diagnosed less than about 5 years ago, wherein the derivative of the LAG-3 protein is the LAG-3Ig fusion protein IMP321.

2. The LAG-3 protein or a derivative thereof for use according to claim 1, wherein the cancer is hormone receptor-positive breast cancer or hormone receptor-negative breast cancer.

3. The LAG-3 protein or derivative thereof for use according to claim 1, wherein the LAG-3 protein or derivative thereof is administered before, during, or after administration of a chemotherapeutic agent, or is administered after administration of a chemotherapeutic agent.

4. 4. The LAG-3 protein or derivative thereof for use according to claim 3, wherein said chemotherapeutic agent is a taxane, optionally paclitaxel.

5. 2. The LAG-3 protein or derivative thereof for use according to claim 1, wherein the subject has one or more of a low monocyte count, has not previously been treated with taxane chemotherapy, has an elevated neutrophil-to-lymphocyte ratio, and was diagnosed less than about 5 years ago, and / or the subject has not previously been treated with taxane chemotherapy and has an elevated neutrophil-to-lymphocyte ratio.

6. 1. A LAG-3 protein or derivative thereof capable of binding to an MHC class II molecule for use in preventing, treating, or ameliorating metastatic breast cancer in a subject, wherein said LAG-3 protein derivative is the LAG-3Ig fusion protein IMP321, and is administered to said subject at a dosage of >30 mg to 200 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

7. 7. The LAG-3 protein or derivative thereof for use according to claim 6, wherein the LAG-3 protein or derivative is administered to the subject at a dose of >30 mg to <120 mg of the molar equivalent of IMP321, at a dose of 50 to 100 mg of the molar equivalent of IMP321, or at a dose of 90 mg of the molar equivalent of IMP321.

8. The LAG-3 protein or derivative thereof for use according to claim 6, wherein multiple doses of the LAG-3 protein or derivative are administered to the subject.

9. 7. The LAG-3 protein or derivative thereof for use according to claim 6, wherein the LAG-3 protein or derivative is administered to the subject before, during, or after administration of a chemotherapeutic agent, and optionally, multiple doses of the chemotherapeutic agent are administered to the subject.

10. The LAG-3 protein or derivative thereof for use according to claim 9, wherein the LAG-3 protein or derivative is administered to the subject on the same day as the chemotherapeutic agent.

11. 10. The LAG-3 protein or derivative thereof for use according to claim 9, wherein the LAG-3 protein or derivative is administered to the subject one or more times in the absence of a chemotherapeutic agent, after which one or more doses of the LAG-3 protein or derivative are administered to the subject before, with, or after one or more doses of the chemotherapeutic agent.

12. 12. The LAG-3 protein or derivative thereof for use according to claim 11, wherein the LAG-3 protein or derivative is administered to the subject on days 1 and 15 of a four-week cycle for up to nine four-week cycles, in the absence of a chemotherapeutic agent. (i) the LAG-3 protein or derivative thereof is administered to the subject over four to eight, preferably six, four-week cycles, in the absence of a chemotherapy agent, following a dose of the LAG-3 protein or derivative on days 1 and 15 of each four-week cycle and a dose of the chemotherapy agent on days 1, 8, and 15 of each four-week cycle; (ii) the LAG-3 protein or derivative thereof is administered to the subject for more than six four-week cycles in the absence of a chemotherapeutic agent following a dose of the LAG-3 protein or derivative on days 1 and 15 of the four-week cycle and a dose of the chemotherapeutic agent on days 1, 8, and 15 of the four-week cycle; or (iii) the LAG-3 protein or derivative thereof is administered to the subject in the absence of a chemotherapeutic agent following a dose of the LAG-3 protein or derivative on days 1 and 15 of the 4-week cycle and a dose of the chemotherapeutic agent on days 1, 8, and 15 of the 4-week cycle for up to thirteen 4-week cycles; The LAG-3 protein or a derivative thereof for use according to claim 11 or 12.

14. 1. A LAG-3 protein or a derivative thereof capable of binding to an MHC class II molecule for use in preventing, treating, or ameliorating metastatic breast cancer in a subject, wherein the LAG-3 protein derivative is the LAG-3Ig fusion protein IMP321 and is administered to the subject on the same day as a chemotherapy agent.

15. 15. The LAG-3 protein or derivative thereof for use according to claim 14, wherein the LAG-3 protein or derivative is administered to the subject on days 1 and 15 of a 4-week cycle, and the chemotherapeutic agent is administered to the subject on days 1, 8, and 15 of the 4-week cycle, and optionally, (i) the 4-week cycle is repeated for 4 to 8 cycles, preferably 6 cycles; (ii) the 4-week cycle is repeated for more than 6 cycles; or (iii) the 4-week cycle is repeated for up to 13 cycles.

16. 1. A LAG-3 protein or derivative thereof capable of binding to an MHC class II molecule for use in preventing, treating, or ameliorating metastatic breast cancer in a subject, wherein the LAG-3 protein derivative is the LAG-3Ig fusion protein IMP321, and the LAG-3 protein or derivative is administered to the subject one or more times in the absence of a chemotherapeutic agent after one or more doses of the LAG-3 protein or derivative have been administered to the subject before, with, or after one or more doses of the chemotherapeutic agent.

17. 17. The LAG-3 protein or derivative thereof for use according to claim 16, wherein the LAG-3 protein or derivative is administered to the subject on days 1 and 15 of a four-week cycle for up to nine four-week cycles, in the absence of a chemotherapeutic agent.

18. (i) the LAG-3 protein or derivative is administered to the subject over four to eight, preferably six, four-week cycles, in the absence of a chemotherapeutic agent, following a dose of LAG-3 protein or derivative on days 1 and 15 of each four-week cycle and a dose of the chemotherapeutic agent on days 1, 8, and 15 of each four-week cycle; (ii) the LAG-3 protein or derivative thereof is administered to the subject for more than six four-week cycles in the absence of a chemotherapeutic agent following a dose of the LAG-3 protein or derivative on days 1 and 15 of the four-week cycle and a dose of the chemotherapeutic agent on days 1, 8, and 15 of the four-week cycle; or (iii) the LAG-3 protein or derivative thereof is administered to the subject in the absence of a chemotherapeutic agent following a dose of the LAG-3 protein or derivative on days 1 and 15 of the 4-week cycle and a dose of the chemotherapeutic agent on days 1, 8, and 15 of the 4-week cycle for up to thirteen 4-week cycles; The LAG-3 protein or a derivative thereof for use according to claim 16 or 17.

19. 17. The LAG-3 protein or derivative thereof for use according to claim 14 or 16, wherein the LAG-3 protein or derivative is administered at a dosage of 6 mg to <120 mg, preferably 20 mg to 100 mg, more preferably 30 mg to 90 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

20. 17. The LAG-3 protein or derivative thereof for use according to claim 9, 14 or 16, wherein said chemotherapeutic agent is administered in a dosage according to approved prescribing information.

21. 17. The LAG-3 protein or derivative thereof for use according to claim 9, 14 or 16, wherein said chemotherapeutic agent is a taxane, optionally paclitaxel.

22. 80 mg / m ２ 17. The method of claim 9, 14, or 16, wherein 100 mg of paclitaxel is administered intravenously to the subject on days 1, 8, and 15 of a 4-week cycle, followed by 90 mg of IMP321 administered subcutaneously on days 1 and 15 of a 4-week cycle.

23. 17. The LAG-3 protein or derivative thereof for use according to claim 1, 6, 14 or 16, wherein the subject is a hormone receptor positive HER2-negative / low (HR+ / HER2-negative / low) metastatic breast cancer patient or a metastatic triple-negative breast cancer (TNBC) patient.

24. 1. A LAG-3 protein or a derivative thereof capable of binding to an MHC class II molecule for use in preventing, treating, or ameliorating metastatic breast cancer in a subject, wherein the LAG-3 protein derivative is the LAG-3Ig fusion protein IMP321, and the subject is a hormone receptor-positive, HER2-negative / low (HR+ / HER2-negative / low) metastatic breast cancer patient or a metastatic triple-negative breast cancer (TNBC) patient.

25. A LAG-3 protein or a derivative thereof capable of binding to an MHC class II molecule for use in preventing, treating, or ameliorating cancer in a subject, wherein the LAG-3 protein derivative is LAG-3Ig fusion protein IMP321 and is administered to the subject at a dose of >30 mg to 200 mg of the molar equivalent of the LAG-3 derivative LAG-3Ig fusion protein IMP321.

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