Compositions and methods for treating macrophage activation syndrome

Abstract

The present disclosure provides compositions and methods for the treatment and prevention of Macrophage Activation Syndrome (MAS) or Hemophagocytic Lymphohistiocytosis (HLH). The methods involve administering to a patient in need thereof an antibody or fragment thereof specific for human GM-CSF protein.

Description

Background Technology

[0001] Hemophagocytic lymphohistiocytosis (HLH) is a fatal autosomal recessive disorder that manifests in early childhood. This disease is characterized by fever, hepatosplenomegaly, cytopenia, and widespread infiltration of vital organs by activated lymphocytes and macrophages. Macrophage activation syndrome (MAS) (also known as secondary HLH) is a rare but serious and life-threatening condition. MAS is considered to be an excessively inflammatory 'cytokine storm' state associated with rheumatic diseases or other triggers. The exact mechanism of MAS is unclear. However, regardless of the initiating events, the final condition is defined by an overactive immune system leading to chronic inflammation and hemophagocytic activity. MAS is characterized by fever, hyperferritinemia, hemophagocytosis, cytopenia (including pancytopenia), and hepatosplenomegaly.

[0002] Current treatments for HLH / MAS rely on various immunosuppressive regimens, including dexamethasone or other steroids, cyclosporine, and etoposide. These therapies are inherently toxic and increase mortality in pre-existing patients. Therefore, there is a strong need for newer, safer treatments for HLH / MAS patients. Summary of the Invention

[0003] This disclosure provides compositions and methods for the treatment and prevention of hemophagocytic lymphohistiocytosis (HLH) or macrophage activation syndrome (MAS). The method involves administering to a patient in need an antibody or fragment thereof specific to the human GM-CSF protein, such as IMH001. Treatment with the anti-GM-CSF antibody provided herein has shown effective efficacy in delaying disease progression.

[0004] One embodiment of this disclosure provides a method for treating or preventing macrophage activation syndrome (MAS) or hemophagocytic lymphohistiocytosis (HLH) in patients of need, the method comprising administering to the patient an antibody or a fragment thereof that is specific to human GM-CSF protein.

[0005] One embodiment of this disclosure provides an antibody or fragment thereof that is specific to human GM-CSF protein for use in the treatment or prevention of macrophage activation syndrome (MAS) or hemophagocytic lymphohistiocytosis (HLH) in patients of need.

[0006] One embodiment of this disclosure provides the use of an antibody or fragment thereof specific to human GM-CSF protein in the preparation of a medicament for the treatment or prevention of macrophage activation syndrome (MAS) or hemophagocytic lymphohistiocytosis (HLH) in patients of need.

[0007] In some embodiments, the antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), the heavy chain variable region comprising the amino acid sequence of SEQ ID NO:1 and the light chain variable region comprising the amino acid sequence of SEQ ID NO:2. In some embodiments, the antibody comprises human IgG1 Fc.

[0008] In some embodiments, the patient has a disease or condition selected from the group consisting of: systemic juvenile idiopathic arthritis (SoJIA), systemic lupus erythematosus (SLE), Kawasaki disease, and adult-onset Still's disease.

[0009] In some embodiments, the patient has a blood ferritin level of at least 300 ng / ml, or at least 350 ng / ml, 400 ng / ml, 450 ng / ml, 500 ng / ml, 550 ng / ml, 600 ng / ml, 650 ng / ml, 684 ng / ml, 700 ng / ml, 720 ng / ml, 740 ng / ml, 750 ng / ml, 760 ng / ml, 784 ng / ml, 800 ng / ml, 820 ng / ml, 840 ng / ml, 850 ng / ml, 860 ng / ml, 884 ng / ml, 900 ng / ml, 950 ng / ml, or 1000 ng / ml.

[0010] In some embodiments, the patient has a hemoglobin level of less than 11 g / dL, or less than 10.5 g / dL, 10 g / dL, 9.5 g / dL, 9 g / dL, 8.5 g / dL, 8 g / dL, 7.5 g / dL, or 7 g / dL.

[0011] In some embodiments, the patient has a platelet count of less than 140 × 10⁻⁶. 9 / L, or less than 130×10 9 / L, 120×10 9 / L、110×10 9 / L, 100×10 9 / L, 90×10 9 / L, 80×10 9 / L, 70×10 9 / L or 60×10 9 / L.

[0012] In some embodiments, the patient has a neutrophil count of less than 1.4 × 10⁻⁶. 9 / L, or less than 1.3×10 9 / L, 1.2×10 9 / L, 1.1×10 9 / L, 1.0×10 9 / L, 0.9×10 9 / L, 0.8×10 9 / L, 0.7×10 9 / L or 0.6×10 9 / L.

[0013] In some embodiments, the patient has a fasting triglyceride level greater than 2.49 mmol / L, or greater than 2.55 mmol / L, 2.6 mmol / L, 2.66 mmol / L, 2.72 mmol / L, 2.77 mmol / L, 2.83 mmol / L, 2.89 mmol / L, 2.94 mmol / L, 3 mmol / L, 3.06 mmol / L, 3.11 mmol / L, 3.17 mmol / L, 3.23 mmol / L, 3.28 mmol / L, 3.34 mmol / L, or 3.4 mmol / L.

[0014] In some embodiments, the patient has a blood fibrinogen level of less than 1.5 g / L, or less than 1.4 g / L, 1.3 g / L, 1.2 g / L, 1.1 g / L, 1.0 g / L, 0.9 g / L, 0.8 g / L, 0.7 g / L, 0.6 g / L, or 0.5 g / L.

[0015] In some embodiments, patients have elevated levels of soluble CD163 compared to healthy individuals. In some embodiments, patients have elevated levels of soluble IL-2 receptors compared to healthy individuals.

[0016] In some embodiments, the antibody or its antigen-binding fragment is administered at a dose of 0.3 mg / kg to 10 mg / kg. In some embodiments, the antibody or its antigen-binding fragment is administered once weekly, once every two weeks, or once monthly. In some embodiments, the antibody or its antigen-binding fragment is administered intravenously or subcutaneously.

[0017] In some embodiments, the antibody is provided in the form of a formulation comprising: 20 mg / ml to 200 mg / ml of antibody; 10 mM to 30 mM of buffer; 130 mM to 250 mM of isotonic regulator; and 0.01% (w / v) to 0.03% (w / v) of surfactant, wherein the formulation has a pH of 4.5 to 7.5.

[0018] In some embodiments, the preparation comprises 50-150 mg / ml of antibody, 10-20 mM histidine, 200-220 mM sucrose and 0.01-0.03% (w / v) polysorbate 80, with a pH of about 5.5-6.1.

[0019] In some embodiments, the preparation comprises about 100 mg / ml of antibody, about 20 mM histidine, about 220 mM sucrose and about 0.02% (w / v) polysorbate 80, with a pH of about 5.8.

[0020] In some embodiments, the method further includes administering a glucocorticoid, cyclosporine, or anakinra to the subject. In some embodiments, the glucocorticoid is selected from the group consisting of: cortisol, cortisone, prednisone, prednisolone, methylprednisolone, dexamethasone, betamethasone, triamcinolone, deflazacort, fludrocortisone acetate, deoxycorticosterone acetate, aldosterone, and beclometasone. Attached Figure Description

[0021] Figure 1 The changes in weight in the treatment group are shown.

[0022] Figure 2 The changes in RBC (inset A), HGB (inset B), NEUT% (inset C), and MONO% (inset D) in the treatment group are shown.

[0023] Figure 3 The changes in total bone marrow viable cells in the treatment group are shown.

[0024] Figure 4 The levels of fibrinogen (inset A), triglycerides (inset B), and ferritin (inset C) in the treatment group are shown.

[0025] Figure 5 The histopathological evaluation of hemophagocytic activity (inset A), inflammatory infiltration (inset B), and lymphocyte depletion (inset C) in the treatment group is shown. Detailed Implementation

[0026] Before describing this disclosure in detail, it should be understood that this disclosure is not limited to the specific compositions or biological systems described, which are of course subject to change. It should also be understood that the terminology used herein is for illustrative purposes only and is not intended to be limiting. As used in this specification and the appended claims, the singular forms “a / an” and “the” include plural references unless the context clearly indicates otherwise. Thus, for example, a reference to “molecule” optionally includes combinations of two or more such molecules, etc.

[0027] As used herein, the term "about" refers to a typical range of error for a corresponding value that is readily known to those skilled in the art. References to "about" values ​​or parameters herein include (and describe) embodiments relating to that value or parameter itself. In cases of doubt or where there is no generally accepted understanding in the art regarding the range of error for a particular value or parameter, "about" means ±1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% of that value or parameter.

[0028] It should be understood that the aspects and embodiments described herein include “comprising aspects and embodiments,” “consisting of aspects and embodiments,” and “substantially consisting of aspects and embodiments.”

[0029] As used herein, the term "antibody" is generally intended to refer to an immunoglobulin molecule comprising four polypeptide chains (two heavy (H) chains and two light (L) chains linked together by disulfide bonds) and its polymers (e.g., IgM); however, immunoglobulin molecules consisting only of heavy chains (i.e., lacking light chains) are also included within the definition of the term "antibody." Each heavy chain contains a heavy chain variable region (abbreviated herein as HCVR or VH) and a heavy chain constant region. The heavy chain constant region contains three domains: CH1, CH2, and CH3. Each light chain contains a light chain variable region (abbreviated herein as LCVR or VL) and a light chain constant region. The light chain constant region contains one domain (CL1). The VH and VL regions can be further subdivided into hypervariable regions called complementarity-determining regions (CDRs), within which are scattered more conserved regions called framework regions (FRs). Each VH and VL consists of three CDRs and four FRs, arranged in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

[0030] Unless otherwise specified, as used herein, the term “antibody” should be understood to encompass the complete antibody molecule as well as its antigen-binding fragment. As used herein, the term “antigen-binding portion” or “antigen-binding fragment” (or simply “antibody portion” or “antibody fragment”) of an antibody refers to one or more fragments of an antibody, such as F(ab’)2, F(ab)2, Fab’, Fab, Fv, scFv, etc., that retain the ability to specifically bind to human GM-CSF or its epitopes.

[0031] The term "specific binding" refers to the formation of a relatively stable complex between an antibody or its antigen-binding fragment and the antigen under physiological conditions. Specific binding can occur through at least approximately 1 × 10⁻⁶ cells / units. -8 The characterization is based on a dissociation constant of M or greater. Methods for determining whether two molecules specifically bind are well-known in the art, including, for example, equilibrium dialysis, surface plasmon resonance, etc. However, isolated antibodies that specifically bind human GM-CSF may be cross-reactive with other antigens, such as GM-CSF from other species (orthogonal homologs). In the context of this disclosure, multispecific (e.g., bispecific) antibodies that bind human GM-CSF and one or more other antigens are considered to "specifically bind" human GM-CSF. Furthermore, the isolated antibodies may be substantially free of other cellular material or chemicals.

[0032] The term "pharmaceutical formulation" refers to a preparation that is in a form in which the biological activity of the active ingredient is permitted and that does not contain any other components that would have unacceptable toxicity to a subject administering the formulation. Such formulations are sterile.

[0033] As used herein, the term "treatment" refers to both therapeutic treatment and preventative or preventative measures aimed at preventing or slowing (alleviating) physiological changes or conditions, such as the progression of cancer. Beneficial or desired clinical outcomes include, but are not limited to, relief of symptoms, reduction of disease severity, stabilization (i.e., non-deterioration) of the disease state, delay or slowing of disease progression, improvement or alleviation of the disease state, and mitigation (whether partial or complete), whether detectable or undetectable. "Treatment" can also refer to extended survival compared to expected survival without treatment. Those in need of treatment include those who already have a condition or symptom, those who are susceptible to a condition or symptom, or those whose condition or symptom requires prevention.

[0034] As used herein, the terms “prevention” or “prophylaxis” refer to preventive or protective treatment of a disease or disease state. Prevention of a disease or disease state may include reducing (e.g., alleviating) one or more symptoms of the disease or disease state, for example, relative to a reference level (e.g., symptoms in a similar subject who has not received treatment). Prevention may also include delaying the onset of one or more symptoms of the disease or disease state, for example, relative to a reference level (e.g., the onset of one or more symptoms in a similar subject who has not received treatment). In embodiments, the disease is the one described herein.

[0035] The terms "subject," "individual," "animal," "patient," or "mammal" refer to any subject who requires diagnosis, prognosis, or treatment, particularly mammalian subjects. Mammal subjects include humans, domestic animals, farm animals and zoo animals, racing animals, or pet animals such as dogs, cats, guinea pigs, rabbits, rats, mice, horses, cattle, cows, etc.

[0036] Treatment of macrophage activation syndrome (MAS) or hemophagocytic lymphohistiocytosis (HLH)

[0037] The inventors of this invention tested the anti-GM-CSF antibody IMH001 against macrophage activation syndrome (MAS) or hemophagocytic lymphohistiocytosis (HLH) in animal models. The animals with MAS were immunodeficient and expressed human GM-CSF and IL-3 cytokines. The animals exhibited progressive disease characteristics of many MAS cases, rapidly developing progressive anemia and weight loss.

[0038] Compared to the control group, animals treated with IMH001 showed reduced weight loss (or even weight gain), improved anemia (improved red blood cell count, hemoglobin, and hematocrit), and inhibited ferritin increase. Therefore, these in vivo data demonstrate the efficacy of IMH001 in treating MAS or HLH.

[0039] Therefore, according to one embodiment of the present technology, a method for treating or preventing macrophage activation syndrome (MAS) or hemophagocytic lymphohistiocytosis (HLH) is provided. In some embodiments, the method involves administering an anti-GM-CSF antibody or antigen-binding fragment to a patient in need. As used herein, the term "GM-CSF" refers to human granulocyte-macrophage colony-stimulating factor. GM-CSF, also known as colony-stimulating factor 2 (CSF2), is a monomeric glycoprotein secreted by macrophages, T cells, mast cells, NK cells, endothelial cells, and fibroblasts, which functions as a cytokine. Pharmaceutical analogues of naturally occurring GM-CSF are also known as sargramostim and molgramostim. Antibodies against human GM-CSF are described, for example, in WO 2006122797, WO 2015028657, and WO2018050111.

[0040] In some embodiments, the anti-GM-CSF antibody is IMH001, which is a humanized IgG1 antibody comprising the heavy chain variable region (VH) of SEQ ID NO:1 and the light chain variable region (VL) of SEQ ID NO:2.

[0041] In some embodiments, the patient has MAS or HLH. In some embodiments, the patient is at risk of developing MAS or HLH. In some embodiments, the HLH / MAS patient or the patient at risk of developing HLH / MAS has a disease or condition such as systemic juvenile idiopathic arthritis (SoJIA), systemic lupus erythematosus (SLE), Kawasaki disease, or adult-onset Still's disease.

[0042] "SoJIA (or the juvenile form of Still's disease)" is a type of juvenile idiopathic arthritis (JIA) that, in addition to arthritis, also presents with extra-articular manifestations such as fever and rash. It was originally called soJIA or Still's disease. The main extra-articular manifestations include high fever, rheumatic rash, hepatosplenomegaly, lymphadenopathy, and anemia. Other manifestations include inflammation of the pleura, pericardium, and cardiac muscle tissue, and inflammation of the peritoneum is also visible.

[0043] Systemic lupus erythematosus (SLE) is an autoimmune disease in which the body's immune system mistakenly attacks healthy tissues in many parts of the body. Symptoms can range from mild to severe and vary from person to person. Common symptoms include joint pain and swelling, fever, chest pain, hair loss, mouth ulcers, swollen lymph nodes, fatigue, and a red rash most commonly seen on the face. There are typically disease phases known as the flare phase and periods of remission with few symptoms.

[0044] Kawasaki disease is a syndrome of unknown cause that causes fever and primarily affects children under 5 years of age. It is a form of vasculitis in which blood vessels throughout the body become inflamed. Fever usually lasts longer than five days and is unresponsive to common medications. Other common symptoms include enlarged lymph nodes in the neck, rashes on the genitals, lips, palms, or soles of the feet, and red eyes. Skin on the hands and feet may peel off within three weeks of onset, after which it usually heals. In some children, coronary artery aneurysms may form in the heart.

[0045] Adult-onset Still's disease (AOSD) is a rare, systemic autoinflammatory disorder characterized by the classic triad of symptoms: fever, joint pain, and a prominent salmon-colored, raised rash. This disease is considered a diagnosis of exclusion. The level of the iron-binding protein ferritin may be dramatically elevated in this condition.

[0046] In some embodiments, the patient has abnormally high blood ferritin levels. In some embodiments, the blood ferritin level is at least 300 ng / ml, or at least 350 ng / ml, 400 ng / ml, 450 ng / ml, 500 ng / ml, 550 ng / ml, 600 ng / ml, 650 ng / ml, 684 ng / ml, 700 ng / ml, 720 ng / ml, 740 ng / ml, 750 ng / ml, 760 ng / ml, 784 ng / ml, 800 ng / ml, 820 ng / ml, 840 ng / ml, 850 ng / ml, 860 ng / ml, 884 ng / ml, 900 ng / ml, 950 ng / ml, or 1000 ng / ml.

[0047] In some embodiments, administration of the anti-GM-CSF antibody provided herein reduced blood ferritin levels by at least 10%, at least 20%, at least 30%, at least 50%, or at least 70% compared to untreated controls. In some embodiments, administration of the anti-GM-CSF antibody provided herein reversed abnormally high blood ferritin levels in patients.

[0048] In some embodiments, the patient has an abnormally low hemoglobin level. In some embodiments, the hemoglobin level is below 11 g / dL, or below 10.5 g / dL, 10 g / dL, 9.5 g / dL, 9 g / dL, 8.5 g / dL, 8 g / dL, 7.5 g / dL, or 7 g / dL.

[0049] In some embodiments, administration of the anti-GM-CSF antibody provided herein increased blood hemoglobin levels by at least 0.2, 0.3, 0.4, 0.5, 1, 1.5, 2, 3, 4, 5, 6, 7, or 8 times compared to untreated controls. In some embodiments, administration of the anti-GM-CSF antibody provided herein reversed abnormally low hemoglobin levels in patients.

[0050] In some embodiments, the patient has an abnormally low platelet count. In some embodiments, the platelet count is below 140 × 10⁻⁶. 9 / L, or less than 130×10 9 / L, 120×10 9 / L、110×10 9 / L, 100×10 9 / L, 90×10 9 / L, 80×10 9 / L, 70×10 9 / L or 60×10 9 / L.

[0051] In some embodiments, administration of the anti-GM-CSF antibody provided herein increased platelet count by at least 0.2, 0.3, 0.4, 0.5, 1, 1.5, 2, 3, 4, 5, 6, 7, or 8-fold compared to untreated controls. In some embodiments, administration of the anti-GM-CSF antibody provided herein reversed abnormally low platelet counts in patients.

[0052] In some embodiments, the patient has an abnormally low neutrophil count. In some embodiments, the neutrophil count is less than 1.4 × 10⁻⁶. 9 / L, or less than 1.3×10 9 / L, 1.2×10 9 / L, 1.1×10 9 / L, 1.0×10 9 / L, 0.9×10 9 / L, 0.8×10 9 / L, 0.7×10 9 / L or 0.6×10 9 / L.

[0053] In some embodiments, administration of the anti-GM-CSF antibody provided herein increased the neutrophil count by at least 0.1, 0.2, 0.3, 0.4, 0.5, 1, 1.5, 2, 3, 4, 5, 6, 7, or 8-fold compared to an untreated control. In some embodiments, administration of the anti-GM-CSF antibody provided herein reversed abnormally low neutrophil counts in patients.

[0054] In some embodiments, the patient has abnormally high fasting triglyceride levels. In some embodiments, fasting triglyceride levels are higher than 2.49 mmol / L, or higher than 2.55 mmol / L, 2.6 mmol / L, 2.66 mmol / L, 2.72 mmol / L, 2.77 mmol / L, 2.83 mmol / L, 2.89 mmol / L, 2.94 mmol / L, 3 mmol / L, 3.06 mmol / L, 3.11 mmol / L, 3.17 mmol / L, 3.23 mmol / L, 3.28 mmol / L, 3.34 mmol / L, or 3.4 mmol / L. In some embodiments, the fasting triglyceride level is higher than 220 mg / dl, or higher than 225 mg / dl, 230 mg / dl, 235 mg / dl, 240 mg / dl, 245 mg / dl, 250 mg / dl, 255 mg / dl, 260 mg / dl, 265 mg / dl, 270 mg / dl, 275 mg / dl, 280 mg / dl, 285 mg / dl, 290 mg / dl, 295 mg / dl, or 300 mg / dl.

[0055] In some embodiments, administration of the anti-GM-CSF antibody provided herein reduced fasting triglyceride levels by at least 10%, at least 20%, at least 30%, at least 50%, or at least 70% compared to untreated controls. In some embodiments, administration of the anti-GM-CSF antibody provided herein reversed abnormally high fasting triglyceride levels in patients.

[0056] In some embodiments, the patient has abnormally low blood fibrinogen levels. In some embodiments, the blood fibrinogen level is below 1.5 g / L, or below 1.4 g / L, 1.3 g / L, 1.2 g / L, 1.1 g / L, 1.0 g / L, 0.9 g / L, 0.8 g / L, 0.7 g / L, 0.6 g / L, or 0.5 g / L.

[0057] In some embodiments, administration of the anti-GM-CSF antibody provided herein increased blood fibrinogen levels by at least 0.1, 0.2, 0.3, 0.4, 0.5, 1, 1.2, 1.5, 2, 3, 4, 5, 6, 7, or 8 times. In some embodiments, administration of the anti-GM-CSF antibody provided herein reversed abnormally low blood fibrinogen levels in patients.

[0058] In some embodiments, administration of the anti-GM-CSF antibody provided herein reduces or reverses the patient’s high level of hemophagocytic activity.

[0059] In some embodiments, administration of the anti-GM-CSF antibody provided herein reduced or reversed the patient’s high level of inflammatory infiltration.

[0060] In some embodiments, administration of the anti-GM-CSF antibody provided herein reduced or reversed patients’ high lymphocyte depletion.

[0061] In some embodiments, administration of the anti-GM-CSF antibody provided herein increases bone marrow viable cells.

[0062] In some embodiments, the patient has macrophage activation. In some embodiments, the patient has elevated levels of soluble CD163 compared to a healthy individual. In some embodiments, the patient has lymphocyte activation. In some embodiments, the patient has elevated levels of soluble IL-2 receptors compared to a healthy individual.

[0063] In some embodiments, the patient meets two or more of the above criteria. In some embodiments, the patient meets three, four, five or more of the above criteria.

[0064] The amount of antibody disclosed herein that is effective in treatment. The precise dose to be used in the formulation will also depend on the route of administration and the severity of the disease, symptoms, or condition, and should be determined based on the physician's judgment and each patient's situation. The effective dose can be deduced from dose-response curves derived from in vitro or animal model testing systems.

[0065] As a general recommendation, the dose of the antigen-binding peptide disclosed herein administered to patients is typically between 0.1 mg / kg and 100 mg / kg of patient body weight, between 0.1 mg / kg and 20 mg / kg of patient body weight, or between 1 mg / kg and 10 mg / kg of patient body weight. Generally, due to the immune response to exogenous peptides, human antibodies have a longer half-life in the human body than antibodies from other species. Therefore, the dose of human antibodies can usually be reduced and the frequency of administration decreased. Furthermore, the dosage and frequency of administration of the antibodies disclosed herein can be reduced by modifications (e.g., lipidation) to enhance antibody uptake and tissue penetration (e.g., into the brain).

[0066] In some embodiments, the dosage of IMH001 is 0.1 to 25 mg / kg, 0.3 to 20 mg / kg, 0.3 to 15 mg / kg, 0.3 to 10 mg / kg, 0.5 to 20 mg / kg, 1 to 20 mg / kg, 5 to 20 mg / kg, 6 to 20 mg / kg, 6 to 10 mg / kg, or 5 to 10 mg / kg per administration. In some embodiments, the dosage is at least 0.3 mg / kg, or at least 0.6 mg / kg, 1 mg / kg, 1.3 mg / kg, 1.6 mg / kg, 2 mg / kg, 2.3 mg / kg, 2.6 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, or 10 mg / kg per administration. In some embodiments, the dose is not higher than 15 mg / kg, 14 mg / kg, 13 mg / kg, 12 mg / kg, 11 mg / kg, 10 mg / kg, 9 mg / kg, 8 mg / kg, 7 mg / kg, 6 mg / kg, 5 mg / kg, 4 mg / kg, 3 mg / kg, or 2 mg / kg. In some embodiments, the administered dose is 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, 10 mg / kg, 12 mg / kg, 15 mg / kg, 20 mg / kg, or 25 mg / kg.

[0067] In some embodiments, the application is performed once a day, once every two days, once every three days, once every four days, once a week, twice a week, once every two weeks, once every three weeks, once a month, or once every two months.

[0068] Methods of administering antibodies include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. Antibodies or compositions can be administered via any convenient route, such as by infusion or bolus injection, absorption through epithelial or mucosal linings (e.g., oral mucosa, rectal and intestinal mucosa), and can be administered together with other bioactive agents. Therefore, pharmaceutical compositions containing the antibodies disclosed herein can be administered orally, rectally, parenterally, intracranially, vaginally, intraperitoneally, topically (e.g., by powder, ointment, drops, or transdermal patch), buccally, or as an oral or nasal spray.

[0069] As used in this article, the term “parenteral” refers to administration methods including intravenous, intramuscular, intraperitoneal, intrasternal, subcutaneous, and intra-articular injections and infusions.

[0070] Administration can be systemic or local. Alternatively, it is preferable to introduce the disclosed antibodies into the central nervous system via any suitable route, including intraventricular and intrathecal injection; intraventricular injection can be facilitated, for example, by an intraventricular catheter attached to a reservoir (such as an Ommaya reservoir). Pulmonary administration can also be used, for example, by using an inhaler or nebulizer, and formulations containing nebulizers.

[0071] Preferably, the antibodies or compositions disclosed herein are applied topically to the area requiring treatment; this can be achieved, for example, but not limited to, local infusion during surgery, topical application (e.g., in conjunction with postoperative wound dressings), by injection, via catheter, via suppository, or via implantation, said implant being a porous, non-porous, or gel-like material, including membranes such as sialastic membranes or fibers. Preferably, when administering the proteins (including antibodies) disclosed herein, care must be taken to use materials that are not absorbed by the protein.

[0072] In another embodiment, antibodies or compositions can be delivered in vesicles, particularly liposomes (see Langer, 1990, Science 249:1527-1533; Treat et al., Liposomes in the Therapy of Infectious Disease and Cancer, Lopez-Berestein and Fidler (eds.), Liss, New York, pp. 353-365 (1989); Lopez-Berestein, ibid., pp. 317-327; see also ibid.).

[0073] In yet another embodiment, the antibody or composition can be delivered in a controlled release system. In one embodiment, a pump can be used (see Sefton, 1987, CRC Crit. Ref. Biomed. Eng. [CRC Biomedical Engineering Qualification Review] 14:201; Buchwald et al., 1980, Surgery [Surgery] 88:507; Saudek et al., 1989, N. Engl. J. Med. [New England Journal of Medicine] 321:574). In another embodiment, polymeric materials may be used (see Medical Applications of Controlled Release, Langer and Wise (eds.), CRC Press, Boca Raton, Florida (1974); Controlled Drug Bioavailability, Drug Product Design and Performance, Smolen and Ball (eds.), Wiley, New York (1984); Ranger and Peppas, J., 1983, Macromol. Sci. Rev. Macromol. Chem. 23:61; see also Levy et al., 1985, Science 228:190; During et al., 1989, Ann. Neurol. 25:351; Howard et al., 1989, J. Neurosurg. 71:105). In yet another embodiment, the controlled-release system can be placed near the therapeutic target (i.e., the brain), thus requiring only a portion of the systemic dose (see, for example, Good Son, in Medical Applications of Controlled Release, ibid., Vol. 2, pp. 115-138 (1984)). Other controlled-release systems are discussed in Langer's review (1990, Science 249: 1527-1533).

[0074] Combinations and formulations

[0075] In some embodiments, a second dose may be administered to the patient. Non-limiting examples of a second dose include glucocorticoids, cyclosporine, and anaspirin.

[0076] Examples of glucocorticoids include cortisol (hydrocortisone), cortisone, prednisone, prednisolone, methylprednisolone, dexamethasone, betamethasone, triamcinolone, dexamethasone acetate, fludrocortisone acetate, deoxycorticosterone acetate, aldosterone, and beclomethasone.

[0077] The formulated antibodies are preferably substantially pure and, ideally, substantially homogeneous (e.g., free of contaminating proteins). "Substantially pure" antibody means a composition containing at least about 90% antibody by weight based on the total weight of the proteins in the composition, preferably at least about 95% antibody by weight. "Substantially homogeneous" antibody means a composition containing at least about 99% antibody by weight based on the total weight of the proteins in the composition.

[0078] The term "medicinal formulation" refers to a preparation containing an anti-GM-CSF antibody in a form that allows the biological activity of the active ingredient to be effective and does not contain any additional components that would have unacceptable toxicity to the subject to whom the formulation will be administered.

[0079] Formulations can be liquid or contain water. Liquid formulations are aqueous solutions or suspensions prepared in a suitable aqueous solvent, such as water or an aqueous / organic mixture (e.g., a water-alcohol mixture).

[0080] In some embodiments, the liquid pharmaceutical formulation comprises antibodies at concentrations of 20 mg / ml to 200 mg / ml, for example, 30 mg / ml to 200 mg / ml, 40 mg / ml to 200 mg / ml, 50 mg / ml to 200 mg / ml, 60 mg / ml to 200 mg / ml, 70 mg / ml to 200 mg / ml, 80 mg / ml to 200 mg / ml, 90 mg / ml to 200 mg / ml, 100 mg / ml to 200 mg / ml, 110 mg / ml to 200 mg / ml. 200mg / ml, 120mg / ml-200mg / ml, 130mg / ml-200mg / ml, 140mg / ml-200mg / ml, 150mg / ml-200mg / ml, 160mg / ml-20 0mg / ml, 170mg / ml-200mg / ml, 180mg / ml-200mg / ml, 190mg / ml-200mg / ml, 20mg / ml-190mg / ml, 20mg / ml-180mg / ml l, 20mg / ml-170mg / ml, 20mg / ml-160mg / ml, 20mg / ml-150mg / ml, 20mg / ml-140mg / ml, 20mg / ml-130mg / ml, 20mg / ml-120mg / ml, 20mg / ml-110mg / ml, 20mg / ml-100mg / ml, 20mg / ml-90mg / ml, 20mg / ml-80mg / ml, 20mg / ml-70mg / ml , 20mg / ml-60mg / ml, 20mg / ml-50mg / ml, 20mg / ml-40mg / ml, 30mg / ml-190mg / ml, 40mg / ml-180mg / ml, 50mg / ml-17 0mg / ml, 60mg / ml-160mg / ml, 70mg / ml-150mg / ml, 80mg / ml-140mg / ml, 90mg / ml-130mg / ml, 100mg / ml-120mg / ml. In some embodiments, the liquid pharmaceutical formulation comprises antibodies at concentrations of 20 mg / ml, 30 mg / ml, 40 mg / ml, 50 mg / ml, 60 mg / ml, 70 mg / ml, 80 mg / ml, 90 mg / ml, 100 mg / ml, 110 mg / ml, 120 mg / ml, 130 mg / ml, 140 mg / ml, 150 mg / ml, 160 mg / ml, 170 mg / ml, 180 mg / ml, 190 mg / ml, or 200 mg / ml.

[0081] In another embodiment, the pharmaceutical formulation further comprises additional excipients. As used herein, the term "excipient" refers to an inert substance commonly used as a diluent, medium, preservative, binder, or stabilizer in pharmaceuticals, which imparts beneficial physical properties to the formulation, such as increased protein stability, increased protein solubility, and reduced viscosity.

[0082] "Excipients" include, but are not limited to: stabilizers, such as human serum albumin (HSA), bovine serum albumin (BSA), α-casein, globulin, α-lactalbumin, LDH, lysozyme, myoglobin, ovalbumin, and RNase A; buffers, such as citric acid, HEPES, PBS, histidine, potassium acetate, potassium citrate, potassium phosphate (KH2PO4), sodium acetate, sodium bicarbonate, sodium citrate, sodium phosphate (NaH2PO4), Tris base, and Tris-HCl; amino acids / metabolites, such as glycine, alanine (α-alanine, β-alanine), arginine, betaine, leucine, lysine, glutamic acid, aspartic acid, histidine, proline, 4-hydroxyproline, sarcosine, γ-aminobutyric acid (GABA), strombine (alanopine, octopine, strombine), and trimethylamine N-oxide (TMAO); and surfactants, such as polysorbate. Ester 20 and polysorbate 80 and poloxamer 407; lipid molecules, such as phosphatidylcholine, ethanolamine and acetyltryptophan ester; polymers, such as polyethylene glycol (PEG) and polyvinylpyrrolidone (PVP) 10, 24, 40; low molecular weight excipients, such as arabinose, cellobiose, ethylene glycol, fructose, fucose, galactose, glycerol / glycerol, glucose, inositol, lactose, maltose, maltotriose, mannose, melibiose, 2-methyl-2,4-pentanediol Octyl ketose, propylene glycol, raffinose, ribose, sucrose, trehalose, xylitol, and xylose; and high molecular weight excipients such as cellulose, β-cyclodextrin, dextran (10 kDa), dextran (40 kDa), dextran (70 kDa), polysucrose, gelatin, hydroxypropyl methylcellulose, hydroxyethyl starch, maltodextrin, methylcellulose, PEG (6 kDa), polydextrose, polyvinylpyrrolidone (PVP) kL5 (10 kDa), PVP (40 kDa), PVP K30 (40kD), PVP K90 (1000kD), dextran gel G-200, and starch; antioxidants such as ascorbic acid, cysteine ​​hydrochloride, thioglycerol, mercaptoacetic acid, thiosorbitol, and glutathione; reducing agents such as cysteine ​​hydrochloride, dithiothreitol, and other thiols or thiophenes; chelating agents such as EDTA, EGTA, glutamic acid, and aspartic acid; inorganic salts / metals such as Ca. 2 +、Ni 2 +, Mg 2 +、Mn 2+, Na2SO4, (NH4)2SO4, Na2HPO4 / NaH2PO4, K2HPO4 / KH2PO4, MgSO4 and NaF; organic salts, such as sodium acetate, sodium polyvinyl chloride, sodium caprylate / Na octanoate, proprionate, lactate, succinate and citrate; organic solvents, such as acetonitrile, dimethyl sulfoxide (DMSO) and ethanol. For further information on excipients, see Remington's Pharmaceutical Sciences (Joseph P. Remington, 18th edition, Mack Publishing Co., Easton, PA), which is incorporated herein by reference in its entirety.

[0083] "Isotonic" means that the target formulation has an osmotic pressure substantially the same as that of human blood. Isotonic formulations typically have an osmotic pressure of about 250 to about 350 mOsm (e.g., about 250 to about 340, about 250 to about 330, about 250 to about 320, about 250 to about 310, about 250 to about 300, about 260 to about 350, about 270 to about 350, about 280 to about 350, about 290 to about 350, about 300 to about 350, about 260 to about 340, about 270 to about 330, about 280 to about 320, about 290 to about 310 mOsm). Isotonic pressure can be measured using, for example, a vapor pressure or a cryogenic osmoremeter. In some embodiments, the isotonic pressure is about 300 mOsm. Isotonic regulators can be one or more selected from the group consisting of sucrose, trehalose, mannitol, arginine, and sodium chloride. In some embodiments, the isotonic modifier ranges from about 130 to 250 mM (e.g., about 130 mM to about 240 mM, about 150 mM to about 240 mM, about 180 mM to about 240 mM, about 200 mM to about 240 mM, about 130 mM to about 220 mM, about 150 mM to about 220 mM, about 180 mM to about 220 mM, about 130 mM to about 210 mM, about 150 mM to about 210 mM, about 180 mM to about 210 mM, about 200 mM to about 220 mM, or about 200 to about 210 mM). In some embodiments, the isotonic regulator is about 130 mM, about 140 mM, about 150 mM, about 160 mM, about 170 mM, about 180 mM, about 190 mM, about 200 mM, about 210 mM, about 220 mM, about 230 mM, about 240 mM, or about 250 mM. In some embodiments, the isotonic regulator comprises sucrose or trehalose.

[0084] As used herein, “pH buffer” refers to a buffered solution that resists pH changes through the action of its acid-base conjugate components. Preferably, the buffer solutions disclosed herein have a pH in the range of about 4.5 to about 7.5 (preferably about 5.0 to about 7.0, for example about 5.0 to about 6.9, about 5.2 to about 6.8, about 5.3 to about 6.7, about 5.4 to about 6.6, about 5.5 to about 6.5, about 5.6 to about 6.4, about 5.7 to about 6.3, about 5.8 to about 6.2, about 5.9 to about 6.1, about 5.5 to about 6.4, about 5.5 to about 6.3, about 5.5 to about 6.2, about 5.5 to about 6.1, or about 5.5 to about 6.0). In some embodiments, the buffer solution has a pH of about 5.0, about 5.1, about 5.2, about 5.3, about 5.4, about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, about 6.0, about 6.1, about 6.2, about 6.3, about 6.4, about 6.5, about 6.6, about 6.7, about 6.8, about 6.9, or 7.0. In one embodiment, the buffer solution has a pH of 5.8. A variety of methods can be used to achieve the desired pH level, including but not limited to adding a suitable buffer solution.

[0085] In some embodiments, the pH buffer contains histidine, acetate, citrate, and succinate. In some embodiments, the pH buffer contains about 10 to about 30 mM histidine and / or about 10 to about 30 mM acetate, such as about 10 to about 25 mM, about 10 to about 20 mM, or about 15 mM to about 20 mM histidine and / or about 10 to about 25 mM, about 10 mM to about 20 mM, or about 15 mM to about 20 mM acetate. In some embodiments, the pH buffer contains about 10 mM, about 11 mM, about 12 mM, about 13 mM, about 14 mM, about 15 mM, about 16 mM, about 17 mM, about 18 mM, about 19 mM, about 20 mM, about 21 mM, about 22 mM, about 23 mM, about 24 mM, about 25 mM, about 26 mM, about 27 mM, about 28 mM, about 29 mM, or about 30 mM of histidine or acetate.

[0086] In some embodiments, the pH buffer comprises L-histidine and histidine hydrochloride monohydrate. In some embodiments, the pH buffer comprises 20 mM histidine, which comprises about 4.4 mM L-histidine and about 15.6 mM histidine hydrochloride monohydrate, about 5.2 mM L-histidine and about 14.8 mM histidine hydrochloride monohydrate, about 6.2 mM L-histidine and about 13.8 mM histidine hydrochloride monohydrate, about 6.8 mM L-histidine and about 13.2 mM histidine hydrochloride monohydrate, about 7.8 mM L-histidine and about 12.2 mM histidine hydrochloride monohydrate, about 8.4 mM L-histidine hydrochloride monohydrate and about 11.6 mM histidine hydrochloride monohydrate, or about 9 mM L-histidine and about 11 mM histidine hydrochloride monohydrate.

[0087] As used herein, “surfactant” refers to a surface-active reagent, preferably a nonionic surfactant. Examples of surfactants described herein include polysorbates (e.g., polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 65, polysorbate 80 and polysorbate 85); poloxamer (e.g., poloxamer 188 and poloxamer 407); Triton; sodium dodecyl sulfate (SDS); sodium lauryl sulfate; sodium octyl glycoside. glycoside); lauryl-sulfobetaine, myristyl-sulfobetaine, linoleyl-sulfobetaine, or stearyl-sulfobetaine; lauryl-sarcosine, myristyl-sarcosine, linoleyl-sarcosine, or stearyl-sarcosine; linoleyl-betaine, myristyl-betaine, or cetyl-betaine; lauramidopropyl-betaine, cocamidopropyl-betaine, linolemidopropyl-betaine, myristamidopropyl-betaine, palmitamidopropyl-betaine, or isostearamidopropyl-betaine (e.g., lauramidopropyl); myristamidopropyl-dimethylamine, palmitamidopropyl-dimethylamine, or isostearamidopropyl-dimethylamine; sodium methylcocoyl taurate or disodium methyloleyl taurate; and MONAQUAT TM Series (Mona Industries, Inc., Paterson, NJ); polyethylene glycol, polypropylene glycol, and copolymers of ethylene glycol and propylene glycol (e.g., Piuronics, PF68, etc.); etc.

[0088] The surfactant concentration is typically from about 0.0001% (w / v) to about 1.0% (w / v), from about 0.01% (w / v) to about 0.5% (w / v), for example, from about 0.015% (w / v) to about 0.03% (w / v), from about 0.02% (w / v) to about 0.03% (w / v), from about 0.025% (w / v) to about 0.03% (w / v), from about 0.01% (w / v) to about 0.025% (w / v), from about 0.01% (w / v) to about 0.02% (w / v), or from about 0.01% (w / v) to about 0.015% (w / v). In one embodiment, the surfactant provided herein comprises polysorbate 80 or polysorbate 20. In some embodiments, the surfactant comprises about 0.01% (w / v), 0.015% (w / v), 0.02% (w / v), 0.025% (w / v), or 0.03% (w / v) of polysorbate 80 or polysorbate 20. In some embodiments, the surfactant comprises about 0.02% (w / v) of polysorbate 80.

[0089] In some embodiments, the liquid pharmaceutical formulation further comprises an antioxidant, a preservative, or a mixture thereof.

[0090] The term "antioxidant" refers to an agent that inhibits the oxidation of other molecules. Examples of antioxidants include ascorbic acid, citrate, lipoic acid, uric acid, cysteine ​​hydrochloride, monothioglycerol, thioglycerol, thioglycolic acid, thiosorbitol, tocopherol, carotene, lycopene, and glutathione; reducing agents, such as cysteine ​​hydrochloride, dithiothreitol; phosphonate compounds, such as etidronic acid, deferoxamine, and malate; and other thiols or thiophenes and methionines. In other embodiments, the antioxidant is a metal chelating agent. Metal chelating agents include, but are not limited to, ethylenediaminetetraacetic acid ("EDTA"), ethylene glycol tetraacetic acid ("EGTA"), thiamine tetrahydrofurfuryldisulfide ("TTFD"), and 2,3-dimercaptosuccinic acid ("DMSA"). In some embodiments, the formulation contains about 1 mM to about 50 mM of antioxidant. In one embodiment, the formulation comprises about 5 mM, about 10 mM, about 15 mM, about 20 mM, about 25 mM, about 30 mM, about 35 mM, about 40 mM, or about 45 mM of antioxidant.

[0091] The term "preservative" refers to a pharmaceutically acceptable excipient that prevents the growth of microorganisms within a composition. More specifically, this disclosure provides a multi-dose liquid composition containing a preservative that protects the composition from microbial contamination.

[0092] In one embodiment, the preservative is present in the composition at an amount of 0.001% (w / v) to 2% (w / v). In one embodiment, the preservative is present in the composition at an amount of 0.002% (w / v) to 1% (w / v). In one embodiment, one or more preservatives are selected from phenol, m-cresol, benzyl alcohol, chlorobutanol, ethanol, phenoxyethanol, p-chloro-m-cresol, methylparaben, propylparaben, benzalkonium chloride, thimerosal, or any combination thereof. In one embodiment, one or more preservatives are selected from phenol, m-cresol, benzyl alcohol, and chlorobutanol.

[0093] The viscosity of anti-GM-CSF antibody formulations can be controlled for subcutaneous, intravenous, or intramuscular administration. Viscosity can be affected by protein concentration and pH. For example, viscosity can increase with increasing protein concentration. Increasing pH can decrease the viscosity of anti-GM-CSF antibody formulations. In some protein formulations, sodium chloride is added to reduce viscosity. Other components that can affect the viscosity of anti-GM-CSF antibody formulations are amino acids, such as histidine and arginine.

[0094] The liquid pharmaceutical formulations described herein can have a variety of viscosities. Methods for measuring the viscosity of liquid pharmaceutical formulations are known to those skilled in the art and may include, for example, a rheometer (e.g., an Anton Paar MCR301 Rheometer with 50 mm, 40 mm, or 20 mm cone fittings). In some embodiments disclosed herein, the viscosity is reported at the high shear limit at a shear rate of 1000 / s. In some embodiments, the liquid pharmaceutical formulation has a viscosity between 1.0 cP ± 10% and 20 cP ± 10%. In some embodiments, the liquid pharmaceutical formulation has a viscosity less than 20 cP, less than 18 cP, less than 15 cP, less than 13 cP, or less than 11 cP. Those skilled in the art will understand that viscosity is temperature-dependent; therefore, unless otherwise stated, the viscosities provided herein are measured at 25°C. In some embodiments, the viscosity of the liquid pharmaceutical preparation is 1.0 cP ± 10%, 2.0 cP ± 10%, 3.0 cP ± 10%, 3.1 cP ± 10%, 3.2 cP ± 10%, 3.5 cP ± 10%, 3.6 cP ± 10%, 3.8 cP ± 10%, 4.0 cP ± 10%, 5.0 cP ± 10%, 5.3 cP ± 10%, 6.0 cP ± 10%, and 6.3 cP at 25°C. ±10%, 6.4cP±10%, 6.8cP±10%, 7.0cP±10%, 7.1cP±10%, 7.4cP±10%, 8.0cP±10%, 9.0cP±10%, 10.0cP±10%, 11.0cP±10%, 12.0cP±10%, 13.0cP±10%, 14.0cP±10%, 15.0cP±10%, or 16cP±10%.

[0095] In some embodiments, the liquid pharmaceutical formulation further comprises a viscosity modifier. In one embodiment, the viscosity modifier is an amino acid. In one embodiment, the viscosity modifier is L-proline. In some embodiments, the concentration of the viscosity modifier is from 1% ± 0.2% to 5% ± 1% w / v. In one embodiment, the viscosity modifier is proline at a concentration of 1.5% ± 0.3% or about 1.5%. In one embodiment, the viscosity modifier is proline at a concentration of 3% ± 0.6% or about 3%.

[0096] In a first aspect, this disclosure provides a novel liquid pharmaceutical formulation comprising:

[0097] a) Anti-GM-CSF antibodies with concentrations of 20 mg / ml to 200 mg / ml were used as antibodies;

[0098] b) Acetate or histidine at a concentration of 10mM-30mM as a buffer;

[0099] c) Sucrose or trehalose at a concentration of 130mM-250mM as an isotonic regulator;

[0100] d) Polysorbate 80 or polysorbate 20 at a concentration of 0.01% (w / v) to 0.03% (w / v) was used as a surfactant;

[0101] The pH of the preparation is from about 4.5 to about 7.5, preferably from about 5.5 to about 6.1.

[0102] In a further embodiment, the formulation does not contain other excipients.

[0103] In a preferred embodiment, this disclosure provides a novel liquid pharmaceutical preparation comprising:

[0104] a) Anti-GM-CSF antibodies at concentrations of 50 mg / ml to 150 mg / ml;

[0105] b) Histidine at a concentration of 10mM-20mM;

[0106] c) Sucrose with a concentration of 200mM-220mM;

[0107] d) Polysorbate 80 at a concentration of 0.01% (w / v) to 0.03% (w / v);

[0108] The formulation has a pH of 5.5 to 6.1.

[0109] In a further embodiment, the formulation does not contain other excipients.

[0110] In a more preferred embodiment, this disclosure provides a novel liquid pharmaceutical formulation comprising:

[0111] a) Anti-GM-CSF antibody at a concentration of 100 mg / ml, wherein the anti-GM-CSF antibody comprises the heavy chain variable region CDR1 (HCDR1) of SEQ ID NO:1, HCDR2 of SEQ ID NO:2, HCDR3 of SEQ ID NO:3, the light chain variable region CDR1 (LCDR1) of SEQ ID NO:4, LCDR2 of SEQ ID NO:5 and LCDR3 of SEQ ID NO:6;

[0112] b) Histidine at a concentration of 20 mM;

[0113] c) Sucrose at a concentration of 220 mM;

[0114] d) Polysorbate 80 at a concentration of 0.02% (w / v);

[0115] The preparation has a pH of 5.8.

[0116] In a further embodiment, the formulation does not contain other excipients.

[0117] The pharmaceutical formulations can be administered to patients via parenteral routes such as injection (e.g., subcutaneous, intravenous, intramuscular, intraperitoneal, etc.) or percutaneously, via mucous membranes, nose, lungs, or orally. Many reusable pen or auto-injector delivery devices can be used for subcutaneous delivery of the pharmaceutical formulations disclosed herein. Examples include, but are not limited to: AUTOPEN TM (Owen Mumford, Inc., Woodstock, UK) DISETRONIC TM Pen (Disetronic Medical Systems, Bergdorf, Switzerland), HUMALOG MIX 75 / 25 TM Pen, HUMALOG TM Pen, HUMALIN 70 / 30 TM Pen (Eli Lilly and Co., Indianapolis, Inc.), NOVOPEN TM I, II and III (Novo Nordisk, Copenhagen, Denmark), NOVOPEN JUNIOR TM (Novo Nordisk, Copenhagen, Denmark), BD TM Pen (BD Company (Becton Dickinson, Franklin Lake, New Jersey), OPTIPEN) TM OPTIPEN PRO TM OPTIPEN STARLET TM and OPTICLIK TM (Sanofi-Aventis, Frankfurt, Germany). Examples of disposable pen or auto-injector delivery devices for subcutaneous delivery of the disclosed pharmaceutical compositions include, but are not limited to: SOLOSTAR TM Pen (Sanofi-Aventis), FLEXPEN TM (Novo Nordisk) and KWIKPEN TM Eli Lilly and Company, SURECLICK TM Automatic injectors (Amgen, Thousand Oaks, California), PENLET TM(Haselmeier, Stuttgart, Germany), EPIPEN (Dey, LP) and HUMIRA TM Pen (Abbott Labs, Abbott Park, Illinois).

[0118] The use of microinfusion devices to deliver the pharmaceutical formulations disclosed herein is also contemplated herein. As used herein, the term “microinfusion device” means a subcutaneous delivery device designed to slowly administer large volumes (e.g., up to about 2.5 mL or more) of a therapeutic formulation over an extended period of time (e.g., about 10, 15, 20, 25, 30 or more minutes). See, for example, U.S. Patent Nos. 6,629,949; 6,659,982; and Meehan et al., J. Controlled Release 46:107-116 (1996). Microinfusion devices are particularly suitable for delivering large doses of therapeutic proteins contained in high concentrations (e.g., about 100, 125, 150, 175, 200 mg / mL or higher) or viscous solutions.

[0119] In some embodiments, this disclosure provides a pre-filled syringe comprising any of the liquid formulations described herein. In some embodiments, the syringe is a 1 mL or 2.25 mL long glass syringe filled with a 27-gauge thin-walled needle, a fluorocarbon-coated rubber plunger, and a rubber needle guard.

[0120] In one respect, the liquid pharmaceutical preparations disclosed herein can be stored at room temperature, refrigerated (e.g., 2-8°C) or frozen (e.g., -20°C or -70°C).

[0121] In some embodiments, the formulation of any of the foregoing aspects has properties selected from the group consisting of: (i) the formulation is stable under long-term storage at 50°C, 40°C, 25°C, 5°C, -20°C, -30°C, and -80°C; (ii) the formulation has low viscosity (viscosity less than 10 cP); (iv) the formulation is isotonic to physiological conditions; (v) the formulation is stable and compatible with intravenous or subcutaneous delivery devices and procedures; and (vi) the formulation is stable under long-term storage in glass vials or pre-filled syringes.

[0122] On the one hand, formulations largely retain their physical and chemical stability and biological activity during storage. Shelf life is typically selected based on the expected shelf life of the formulation. Various analytical techniques for measuring protein stability are available in the field and have been reviewed, for example, in: Peptide and Protein Drug Delivery, 247-301, edited by Vincent Lee, Marcel Dekker, Inc., New York, NY, published (1991) and Jones, A. Adv. Drug Delivery Rev. 10:29-90 (1993). Stability can be measured at a selected temperature over a selected time period. For example, liquid formulations are stable at approximately 40°C for at least approximately 3 days, 5 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, or 6 weeks. On the other hand, the liquid preparation is stable at about 5°C and / or 25°C for at least about 1 month, at least about 3 months, at least about 6 months, at least about 9 months, at least about 12 months, at least about 18 months, at least about 24 months, at least about 30 months, or at least about 36 months; and / or at about -20°C and / or -70°C for at least about 1 month, at least about 3 months, at least about 6 months, at least about 9 months, at least about 12 months, at least about 18 months, at least about 24 months, at least about 30 months, at least about 36 months, at least about 42 months, or at least about 48 months. Furthermore, in some embodiments, the liquid preparation may be stable after freezing (to, for example, -80°C) and thawing (e.g., after 1, 2, or 3 freeze-thaw cycles).

[0123] The stability of liquid formulations can be evaluated qualitatively and / or quantitatively in a variety of ways, including evaluating dimer, multimer, and / or aggregate formation (e.g., using size exclusion chromatography (SEC), matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF MS), analytical ultracentrifugation, light scattering (photon correlation spectroscopy, dynamic light scattering (DLS), static light scattering, multi-angle laser light scattering (MALLS)), and flow-based microscopy). Imaging; electrical impedance (coulter) counting, light obscuration or other liquid particle counting systems, by measuring turbidity and / or by visual inspection; assessing charge heterogeneity by using cation exchange chromatography (CEX), isoelectric focusing (IEF) (e.g., capillary technique (cIEF)) or capillary zone electrophoresis; N-terminal or C-terminal sequence analysis; mass spectrometry; SDS-PAGE or SEC analysis to compare fragmented, intact and multimeric (i.e., dimer, trimer, etc.) antibodies; peptide mapping (e.g., trypsin or LYS-C) analysis; evaluating the biological activity or antigen-binding function of antibodies; and similar methods. The stability of formulations in solid form can also be evaluated qualitatively and / or quantitatively in a variety of ways, including direct testing, such as identifying crystal structures by X-ray powder diffraction (XRPD); evaluating antibody structures in solid form using Fourier transform infrared spectroscopy (FTIR); and measuring thermal transitions (melting, glass transition, etc.) in lyophilized solids using differential scanning calorimetry (DSC) and indirect testing (such as measuring moisture content via the Karl Fischer test), for example, to infer the likelihood of chemical instability caused by hydrolysis. Instability can involve any or more of the following: aggregation (e.g., non-covalent soluble aggregation, covalent soluble aggregation (e.g., disulfide rearrangement / disordering), insoluble aggregation), deamidation (e.g., Asn deamidation), oxidation (e.g., Met oxidation), isomerization (e.g., Asp isomerization), shearing / hydrolysis / fragmentation (e.g., hinge region fragmentation), succinimide formation, N-terminal extension, C-terminal processing, differential glycosylation, etc.

[0124] Example

[0125] Example 1. IMH001 inhibited the development of MAS and improved symptoms.

[0126] This study was conducted to evaluate the in vivo efficacy of the GM-CSF neutralizing antibody IMH001 (see sequence in Table 1) in delaying disease progression in a NOG-EXL mouse model of macrophage activation syndrome (MAS) induced by CBMC transplantation. This lymphocyte-independent effector-driven MAS mouse model can be established by transplanting human umbilical cord blood (UCB) into NOG-EXL mice. NOG-EXL (NOD.Cg-Prkdc) scid IL2rg tm1Sug Tg(SV40 / HTLV-IL3,CSF2) is based on NOG(NOD.Cg-Prkdc) scid IL2rg tm1Sug A genetically modified strain of NOG-EXL mice was used. This strain exhibited immunodeficiency and persistent overexpression of human IL-3 and GM-CSF, and it could enrich myeloid cell populations, including Dc, monocytes, and macrophages, after human stem cell transplantation. A total of 46 NOG-EXL mice and 8 NOG mice were used in this study.

[0127] Table 1. VH / VL sequence of IMH001

[0128]

[0129] This model demonstrates a progressive disease with many characteristics of MAS, rapidly developing into progressive anemia without the need for exogenous immune stimulation. Therefore, this model can be used to examine the contribution of the monocyte / macrophage arm and identify novel therapeutic strategies against MAS / HLH.

[0130] 1.1 Method

[0131] Prior to remodeling of umbilical cord blood mononuclear cells (CBMCs) pretreated with OKT3 via tail vein injection, five- to six-week-old NOG-EXL mice were irradiated with 1 Gy. Remodeling day was defined as day 0. Starting from week 1 after implantation, NOG-EXL mice were treated with OKT3-pretreated CBMCs via intraperitoneal (ip) of PBS or 10 mg / kg IMH001 every two weeks. Animal body weight, CBC count, and serum ferritin were measured at designated time points.

[0132] One week after reconstruction (day 7), four NOG mice (G1) and 14 NOG-EXL mice (6 G2 mice and 8 G3 mice, respectively) were administered the drug twice a week for 15 weeks. The grouping and administration regimens are shown in Table 2 below:

[0133] Table 2. Grouping and Dosing Regimens

[0134]

[0135] Body weight: The body weight of the mice was measured and recorded weekly after CBMC transplantation.

[0136] CBC Count: Whole blood was collected from mice at weeks 4, 8, and 16 and counted using a CBC machine. A total bone marrow viable cell count was also performed at the end of the study, week 16. Mice were anesthetized with 3%–4% isoflurane prior to blood collection.

[0137] Cytokines measured by MSD method: Serum was collected at weeks 8 and 10 for the determination of IL-1β, IL-6, TNF-α, GM-CSF, IFN-γ, and IL-12. Mice were anesthetized with 3%–4% isoflurane prior to blood collection.

[0138] Ferritin, fibrinogen, and triglycerides: Serum was collected from mice at weeks 8 and 16. Measurements were then performed individually using an ELISA kit. Mice were anesthetized with 3%–4% isoflurane prior to blood collection.

[0139] Experimental endpoint collection: The endpoint was day 112 / week 16. Spleens were collected from all mice in each group and weighed; cell smears from bone marrow, liver, and spleen sections were collected for Wright Giemsastaining to detect the presence of hemophagocytic activity. Activated lymphoid tissue, spleen / lymph nodes, liver, and meninges were collected for H&E sections to detect inflammatory cell infiltration and reduced lymphoid follicles in the spleen / lymph nodes.

[0140] statistics

[0141] All statistical analysis methods used in this project must be consistent with the experimental plan, and results are expressed as mean ± SEM. First, the F-test was used (IDBS E-WorkBook 10.5.0 (64-bit)). IDBS (Analysis of Variance) is used to analyze homogeneity of variance. If the variances are homogeneous (p>0.05), a one-way ANOVA is performed. The t-test is used to compare two groups, and Dunnett's multiple comparisons method is used to compare multiple groups. If the variances are heteroscedastic (p≤0.05), the Kruskal-Wallis nonparametric test is used, the U-test is used to compare two groups (Kruskal-Wallis parametric test shows a significant difference, p≤0.05), and Dunn's multiple comparisons method is used for multiple group comparisons (Kruskal-Wallis parametric test shows no significant difference, p>0.05). p<0.05 is considered significant.

[0142] 1.2 Results

[0143] Shortly after OKT3-pretreated CBMCs were implanted into NOG-EXL mice, the mice developed MAS, characterized by weight loss, a gradual decrease in red blood cell (RBC) count, hemoglobin (HGB) and hematocrit (HCT), and high ferritin.

[0144] 1.2.1 Weight Changes

[0145] However, treatment with IMH001 completely reversed the progression of the disease. Animals were weighed weekly after CBMC implantation.

[0146] No obvious abnormalities were found in any group during the experiment. Figure 1 The figure shows the changes in body weight at each time point in each group. Sustained weight loss was observed in NOG or NOG-EXL mice transplanted with CBMCs as the disease progressed. Compared to the vector control group (G2), the body weight of the IMH001 treatment group (G3: 10 mg / kg) significantly increased from day 63 to day 112 (p<0.05). This result indicates that IMH001 treatment effectively improves weight loss induced by MAS in animals.

[0147] 1.2.2 Peripheral blood and bone marrow cell counts

[0148] Whole blood was collected from mice, and whole blood counts (CBCs) were analyzed at weeks 4, 8, 12, and 16. At the end of the study (week 16), total bone marrow viable cells were also counted. Figure 2 The figures show RBC, HGB, NEUT, and MONO at each time point in each group. As the disease progressed, a sustained decrease in peripheral RBC count, HGB level, and NEUT percentage was observed in NOG-EXL:CBMC transplanted mice, while PLT count and MONO percentage did not change significantly. Following IMH001 treatment, a recovery in the decrease in RBC and NEUT was observed, particularly on day 56, with significantly higher RBC count and HGB level compared to untreated mice (6.45 vs. 8.17, p < 0.001). A decrease in the percentage of monocytes was also observed in the IMH001 treatment group, which was attributed to the pharmacological effect of IMH001.

[0149] Figure 3 Total bone marrow viable cells are shown. Compared to NOG-EXL-untreated mice, there was a trend towards an increase in total bone marrow cells at day 112 (mean 9.55 vs 11.6, p = 0.182). The number of peripheral RBCs continued to decrease in the MAS control group. Treatment with IMH001 effectively improved the number of RBCs in peripheral blood, accompanied by an increase in HGB.

[0150] 1.2.3 Ferritin, fibrinogen, and triglycerides

[0151] Serum was collected from mice at weeks 8 and 16. Figure 4 The figures show ferritin, fibrinogen, and triglycerides at each time point in each group. In NOG-EXL CBMC transplanted mice, ferritin, an indicator of macrophage activation, significantly increased with disease progression, a phenomenon not observed in NOG CBMC transplanted mice. Moreover, IMH001 treatment effectively controlled ferritin levels compared to untreated mice, as observed at weeks 8 and 16. No significant changes in fibrinogen and triglycerides were observed during disease progression or after IMH001 treatment.

[0152] The results showed a significant increase in peripheral blood ferritin in the MAS control group, indicating macrophage activation. Treatment with IMH001 significantly reduced peripheral blood ferritin levels.

[0153] 1.2.4 Histopathological evaluation

[0154] At the end of the study (week 16), cell smears from bone marrow, liver, and spleen sections were collected for Wright-Giemsa staining to detect the presence of hemophagocytic activity. Activated lymphoid tissue / lymph nodes and spleen were collected for H&E staining to assess lymphocyte depletion; the liver and brain / meninges were H&E stained to detect inflammatory cell infiltration. Figure 5 The results are shown in the figure.

[0155] Hemophagocytic activity: Macrophages containing hemosiderin were observed in spleen red pulp sections and bone marrow smears from the MAS control group, indicating hemophagocytic activity. IMH001 treatment can effectively prevent MAS-related hemophagocytic activity.

[0156] Lymphocyte depletion: NOG-EXL mice showed a decrease in the number of lymphoid follicles and germinal centers in the spleen and the number of lymphocytes in the lymph nodes, indicating lymphocyte depletion. IMH001 treatment effectively reduced lymphocyte depletion. Furthermore, a reduction in inflammatory infiltration was observed in the liver and meninges after IMH001 treatment.

[0157] The results showed that 16 weeks after CBMC transplantation, NOG-EXL mice exhibited increased hemophagocytic activity, lymphocyte depletion, and elevated inflammation compared to NOG mice. These histopathological parameters were significantly improved in IMH001-treated mice.

[0158] Hemophagocytic function: Macrophages containing hemosiderin were observed in spleen red pulp sections and bone marrow smears of the MAS control group, indicating hemophagocytic function.

[0159] Lymphocyte depletion: NOG-EXL mice showed a decrease in the number of lymphoid follicles and germinal centers in the spleen and the number of lymphocytes in the lymph nodes, indicating lymphocyte depletion.

[0160] This study showed that, compared with tocilizumab, IMH001 treatment demonstrated efficacy in delaying disease progression across all the aforementioned parameters.

[0161] Example 2. Development of IMH001 formulation

[0162] This example demonstrates the development of a suitable formulation for the clinical use of IMH001 in the treatment of MAS.

[0163] First, three different buffer systems were evaluated for IMH001 at various pH levels: sodium acetate, histidine hydrochloride, and sodium dihydrogen phosphate. All three buffer systems were able to stabilize the antibody, and pH 5.5 to 6.0 appeared to be more beneficial than 6.5 or higher. Histidine hydrochloride and pH 5.8 were selected for further testing.

[0164] The next step involved evaluating various excipients, including sucrose, trehalose, mannitol, proline, arginine hydrochloride, glycine, sodium chloride, PS20, and PS80. Formulations with different excipients were compared based on changes in stability (CEX-HPLC) over one week at 50°C. The results showed that sucrose and proline were similar in performance and slightly better than the other formulations. Sucrose was selected for further development.

[0165] However, it is unclear whether the chosen buffer and excipients would make the protein solubility of IMH001 suitable for subcutaneous injection at high concentrations. Therefore, the solubility and viscosity of the protein at concentrations of 100 mg / ml and 150 mg / ml in a histidine hydrochloride buffer system (pH 5.8) were evaluated.

[0166] In a 20 mmol / L histidine hydrochloride buffer system (pH 5.8), when the protein concentration was 100 mg / ml and 150 mg / ml and the samples were placed at 4 °C and 25 °C for 48 hours, no significant changes were observed in appearance, clarity, OD350, protein concentration and viscosity. The viscosity was less than 10 cP and the clarity was less than 6 NTU.

[0167] The ability of PS80 (polysorbate 80) to improve the durability and stability of candidate formulations at a protein concentration of 100 mg / ml was then tested at different sucrose concentrations. The viscosity of all samples was acceptable, and it decreased with decreasing sucrose concentration.

[0168] After further adjustments to the concentration of each component, the following formulation was determined to be superior to other candidate formulations: 100 mg / ml IMH001, 20 mM histidine, 220 mM sucrose, and 0.02% (w / v) polysorbate 80, pH 5.8. This formulation was used as the stock solution for the following in vivo studies.

[0169] ***

[0170] The scope of this disclosure is not limited to the particular embodiments described, which are intended as a single illustration of various aspects of this disclosure, and any functionally equivalent compositions or methods are within the scope of this disclosure. It will be apparent to those skilled in the art that various modifications and variations can be made to the methods and compositions of this disclosure without departing from the spirit or scope of this disclosure. Therefore, this disclosure is intended to cover such modifications and variations, provided they fall within the scope of the appended claims and their equivalents.

[0171] All publications and patent applications mentioned in this specification are incorporated herein by reference to the same extent that each individual publication or patent application is specifically and individually indicated to be incorporated by reference.

Claims

1. Use of an antibody or antigen-binding fragment thereof specific for human GM-CSF protein in the manufacture of a medicament for treating or preventing macrophage activation syndrome in a patient in need thereof, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable region that is the amino acid sequence of SEQ ID NO: 1 and a light chain variable region that is the amino acid sequence of SEQ ID NO:

2.

2. The use of claim 1, wherein the antibody comprises a human IgGl Fc.

3. The use of claim 1 or 2, wherein the patient has a disease or condition selected from the group consisting of systemic-onset juvenile idiopathic arthritis, systemic lupus erythematosus, Kawasaki disease, and adult-onset Still’s disease.

4. The use of claim 1 or 2, wherein the patient has a blood ferritin level of at least 300 ng / ml.

5. The use of claim 1 or 2, wherein the patient has a blood ferritin level of at least 350 ng / ml.

6. The use of claim 1 or 2, wherein the patient has a blood ferritin level of at least 400 ng / ml.

7. The use of claim 1 or 2, wherein the patient has a blood ferritin level of at least 450 ng / ml.

8. The use of claim 1 or 2, wherein the patient has a blood ferritin level of at least 500 ng / ml.

9. The use of claim 1 or 2, wherein the patient has a blood ferritin level of at least 550 ng / ml.

10. The use of claim 1 or 2, wherein the patient has a blood ferritin level of at least 600 ng / ml.

11. The use of claim 1 or 2, wherein the patient has a blood ferritin level of at least 650 ng / ml.

12. The use of claim 1 or 2, wherein the patient has a blood ferritin level of at least 684 ng / ml.

13. The use of claim 1 or 2, wherein the patient has a blood ferritin level of at least 700 ng / ml.

14. The use of claim 1 or 2, wherein the patient has a blood ferritin level of at least 720 ng / ml.

15. The use of claim 1 or 2, wherein the patient has a blood ferritin level of at least 740 ng / ml.

16. The use of claim 1 or 2, wherein the patient has a blood ferritin level of at least 750 ng / ml.

17. The use of claim 1 or 2, wherein the patient has a blood ferritin level of at least 760 ng / ml.

18. The use of claim 1 or 2, wherein the patient has a blood ferritin level of at least 784 ng / ml.

19. The use of claim 1 or 2, wherein the patient has a blood ferritin level of at least 800 ng / ml.

20. The use of claim 1 or 2, wherein the patient has a blood ferritin level of at least 820 ng / ml.

21. The use of claim 1 or 2, wherein the patient has a blood ferritin level of at least 840 ng / ml.

22. The use of claim 1 or 2, wherein the patient has a blood ferritin level of at least 850 ng / ml.

23. The use of claim 1 or 2, wherein the patient has a blood ferritin level of at least 860 ng / ml.

24. The use of claim 1 or 2, wherein the patient has a blood ferritin level of at least 884 ng / ml.

25. The use of claim 1 or 2, wherein the patient has a blood ferritin level of at least 900 ng / ml.

26. The use of claim 1 or 2, wherein the patient has a blood ferritin level of at least 950 ng / ml.

27. The use of claim 1 or 2, wherein the patient has a blood ferritin level of at least 1000 ng / ml.

28. The use of claim 1 or 2, wherein the patient has a hemoglobin level of less than 11 g / dL.

29. The use of claim 1 or 2, wherein the patient has a hemoglobin level of less than 10.5 g / dL.

30. The use of claim 1 or 2, wherein the patient has a hemoglobin level of less than 10 g / dL.

31. The use of claim 1 or 2, wherein the patient has a hemoglobin level of less than 9.5 g / dL.

32. The use of claim 1 or 2, wherein the patient has a hemoglobin level of less than 9 g / dL.

33. The use of claim 1 or 2, wherein the patient has a hemoglobin level of less than 8.5 g / dL.

34. The use of claim 1 or 2, wherein the patient has a hemoglobin level of less than 8 g / dL.

35. The use of claim 1 or 2, wherein the patient has a hemoglobin level of less than 7.5 g / dL.

36. The use of claim 1 or 2, wherein the patient has a hemoglobin level of less than 7 g / dL.

37. The use of claim 1 or 2, wherein the patient has a platelet count of less than 140 x 10 9 / L.

38. The use of claim 1 or 2, wherein the patient has a platelet count of less than 130 x 10 9 / L.

39. The use of claim 1 or 2, wherein the patient has a platelet count of less than 120 x 10 9 / L.

40. The use of claim 1 or 2, wherein the patient has a platelet count of less than 110 x 10 9 / L.

41. The use of claim 1 or 2, wherein the patient has a platelet count of less than 100 x 10 9 / L.

42. The use of claim 1 or 2, wherein the patient has a platelet count of less than 90 x 10 9 / L.

43. The use of claim 1 or 2, wherein the patient has a platelet count of less than 80 x 10 9 / L.

44. The use of claim 1 or 2, wherein the patient has a platelet count of less than 70 x 10 9 / L.

45. The use of claim 1 or 2, wherein the patient has a platelet count of less than 60 x 10 9 / L.

46. The use of claim 1 or 2, wherein the patient has a neutrophil count of less than 1.4 x 10 9 / L.

47. The use of claim 1 or 2, wherein the patient has a neutrophil count of less than 1.3 x 10 9 / L.

48. The use of claim 1 or 2, wherein the patient has a neutrophil count of less than 1.2 x 10 9 / L.

49. The use of claim 1 or 2, wherein the patient has a neutrophil count of less than 1.1 x 10 9 / L.

50. The use of claim 1 or 2, wherein the patient has a neutrophil count of less than 1.0 x 10 9 / L.

51. The use of claim 1 or 2, wherein the patient has a neutrophil count of less than 0.9 x 10 9 / L.

52. The use of claim 1 or 2, wherein the patient has a neutrophil count of less than 0.8 x 10 9 / L.

53. The use of claim 1 or 2, wherein the patient has a neutrophil count of less than 0.7 x 10 9 / L.

54. The use of claim 1 or 2, wherein the patient has a neutrophil count of less than 0.6 x 10 9 / L.

55. The use of claim 1 or 2, wherein the patient has a fasting triglyceride level of more than 2.49 mmol / L.

56. The use of claim 1 or 2, wherein the patient has a fasting triglyceride level of more than 2.55 mmol / L.

57. The use of claim 1 or 2, wherein the patient has a fasting triglyceride level of more than 2.6 mmol / L.

58. The use of claim 1 or 2, wherein the patient has a fasting triglyceride level of more than 2.66 mmol / L.

59. The use of claim 1 or 2, wherein the patient has a fasting triglyceride level of more than 2.72 mmol / L.

60. The use of claim 1 or 2, wherein the patient has a fasting triglyceride level of greater than 2.77 mmol / L.

61. The use of claim 1 or 2, wherein the patient has a fasting triglyceride level of greater than 2.83 mmol / L.

62. The use of claim 1 or 2, wherein the patient has a fasting triglyceride level of greater than 2.89 mmol / L.

63. The use of claim 1 or 2, wherein the patient has a fasting triglyceride level of greater than 2.94 mmol / L.

64. The use of claim 1 or 2, wherein the patient has a fasting triglyceride level of greater than 3 mmol / L.

65. The use of claim 1 or 2, wherein the patient has a fasting triglyceride level of greater than 3.06 mmol / L.

66. The use of claim 1 or 2, wherein the patient has a fasting triglyceride level of greater than 3.11 mmol / L.

67. The use of claim 1 or 2, wherein the patient has a fasting triglyceride level of greater than 3.17 mmol / L.

68. The use of claim 1 or 2, wherein the patient has a fasting triglyceride level of greater than 3.23 mmol / L.

69. The use of claim 1 or 2, wherein the patient has a fasting triglyceride level of greater than 3.28 mmol / L.

70. The use of claim 1 or 2, wherein the patient has a fasting triglyceride level of greater than 3.34 mmol / L.

71. The use of claim 1 or 2, wherein the patient has a fasting triglyceride level of greater than 3.4 mmol / L.

72. The use of claim 1 or 2, wherein the patient has a blood fibrinogen level of less than 1.5 g / L.

73. The use of claim 1 or 2, wherein the patient has a blood fibrinogen level of less than 1.4 g / L.

74. The use of claim 1 or 2, wherein the patient has a blood fibrinogen level of less than 1.3 g / L.

75. The use of claim 1 or 2, wherein the patient has a blood fibrinogen level of less than 1.2 g / L.

76. The use of claim 1 or 2, wherein the patient has a blood fibrinogen level of less than 1.1 g / L.

77. The use of claim 1 or 2, wherein the patient has a blood fibrinogen level of less than 1.0 g / L.

78. The use of claim 1 or 2, wherein the patient has a blood fibrinogen level of less than 0.9 g / L.

79. The use of claim 1 or 2, wherein the patient has a blood fibrinogen level of less than 0.8 g / L.

80. The use of claim 1 or 2, wherein the patient has a blood fibrinogen level of less than 0.7 g / L.

81. The use of claim 1 or 2, wherein the patient has a blood fibrinogen level of less than 0.6 g / L.

82. The use of claim 1 or 2, wherein the patient has a blood fibrinogen level of less than 0.5 g / L.

83. The use of claim 1 or 2, wherein the patient has elevated levels of soluble CD163 compared to a healthy individual.

84. The use of claim 1 or 2, wherein the patient has elevated levels of soluble IL-2 receptor compared to a healthy individual.

85. The use of claim 1 or 2, wherein the antibody or antigen-binding fragment thereof is administered at 0.3 mg / kg to 25 mg / kg.

86. The use of claim 1 or 2, wherein the antibody or antigen-binding fragment thereof is administered at 5 mg / kg to 20 mg / kg.

87. The use of claim 1 or 2, wherein the antibody or antigen-binding fragment thereof is administered at 5 mg / kg to 10 mg / kg.

88. The use of claim 1 or 2, wherein the antibody or antigen-binding fragment thereof is administered at 6 mg / kg to 10 mg / kg.

89. The use of claim 1 or 2, wherein the antibody or antigen-binding fragment thereof is administered once a week, twice a week, once every two weeks, or once a month.

90. The use of claim 1 or 2, wherein the antibody or antigen-binding fragment thereof is administered intravenously or subcutaneously.

91. The use of claim 3, wherein the antibody is provided in a formulation comprising: (a) 20 mg / ml - 200 mg / ml of the antibody; (b) 10 mM - 30 mM of a buffering agent; (c) 130 mM - 250 mM of an isotonicity adjusting agent; and (d) 0.01% w / v - 0.03% w / v of a surfactant, wherein the formulation has a pH of 4.5 - 7.

5.

92. The use of claim 91, wherein the formulation comprises 50 - 150 mg / ml of the antibody, 10 - 20 mM histidine, 200 - 220 mM sucrose, and 0.01 - 0.03% w / v polysorbate 80, at a pH of 5.5 - 6.

1.

93. The use of claim 92, wherein the formulation comprises about 100 mg / ml of the antibody, about 20 mM histidine, about 220 mM sucrose, and about 0.02% w / v polysorbate 80, at a pH of about 5.

8.

94. The use of claim 1 or 2, wherein the treatment or prevention further comprises administering to the patient a glucocorticoid, a cyclosporine, or an anakinra.

95. The use of claim 94, wherein the glucocorticoid is selected from the group consisting of cortisone, cortisone acetate, prednisone, prednisolone, methylprednisolone, dexamethasone, betamethasone, triamcinolone, deflazacort, and beclomethasone.

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