Methods of treating cardiovascular diseases

Abstract

The present disclosure provides a method of treating cardiovascular disease comprising subcutaneously administering to a patient in need thereof a therapeutically effective dose of an anti-interleukin-6 (anti-IL-6) antibody or antibody fragment. Also provided herein are pharmacologically active agents, compositions, methods, and / or dosing schedules for treating cardiovascular diseases.

Description

[0001] Cross - Reference to Related Applications

[0002] This application claims the benefit of priority of U.S. Provisional Patent Application No. 63 / 383,429, filed on November 11, 2022, and U.S. Provisional Patent Application No. 63 / 586,027, filed on September 28, 2023, which are hereby incorporated by reference in their entireties for all purposes.

[0003] Reference to an Electronic Sequence Listing

[0004] The content of the electronic sequence listing (TOUR_004_02WO_SeqList_ST26.xml; size: 14,173 bytes; created on November 3, 2023) is hereby incorporated by reference in its entirety. Technical Field

[0005] This disclosure relates to therapeutic antibody molecules and the treatment of cardiovascular diseases. Background Art

[0006] Cardiovascular diseases are one of the leading causes of death in the United States and most European countries. It is estimated that more than 70 million people in the United States alone suffer from cardiovascular diseases or disorders, including but not limited to hypertension, coronary heart disease, dyslipidemia, congestive heart failure, and stroke.

[0007] Interleukin - 6 (IL - 6) is a widely expressed cytokine that can cause inflammation and oxidative stress, which can further lead to heart damage. Increased interleukin - 6 expression is closely associated with atherosclerosis, myocardial infarction, heart failure, and ischemic stroke. Lipid - lowering is the main treatment for atherosclerosis. However, even in patients with optimal lipid management, the cardiovascular risk remains high.

[0008] IL - 6 inhibition has been demonstrated as a therapeutic strategy to reduce cardiovascular (CV) morbidity and mortality. A phase II clinical trial of the human anti - IL - 6 monoclonal antibody ziltivekimab showed that ziltivekimab significantly reduced various inflammation biomarkers related to atherosclerosis in patients with advanced chronic kidney disease (CKD) (NCT03926117). However, anti - drug antibodies were detected in 12.5% of patients taking 2 mg of ziltivekimab, in 6.3% of patients taking 6 mg of ziltivekimab, and in 12.5% of patients taking 20 mg of ziltivekimab.

[0009] Individuals with moderate chronic kidney disease and elevated IL - 6 or C - reactive protein have a significantly increased risk of major adverse cardiovascular events and represent a large group with unmet treatment needs. SUMMARY OF THE INVENTION

[0010] Provided herein is a method for treating cardiovascular diseases, which comprises administering to a patient in need thereof a therapeutically effective dose of an anti-interleukin-6 (anti-IL-6) antibody or antibody fragment, wherein the antibody or antibody fragment has variable heavy chain (VH) CDRs defined in SEQ ID NOs: 2, 3, and 4, and variable light chain (VL) CDRs defined in SEQ ID NOs: 8, 9, and 10.

[0011] In some embodiments, the anti-IL-6 antibody or antibody fragment comprises a heavy chain polypeptide and a light chain polypeptide, wherein the heavy chain polypeptide comprises a polypeptide having at least about 98% identity to SEQ ID NO: 1, and the light chain polypeptide comprises a polypeptide having at least about 98% identity to SEQ ID NO: 7. In one aspect, the anti-IL-6 antibody or antibody fragment comprises a heavy chain polypeptide having the sequence of SEQ ID NO: 1 and a light chain polypeptide having the sequence of SEQ ID NO: 7.

[0012] In some embodiments, the patient being treated according to the methods of the present disclosure is at least 18 years old.

[0013] In some embodiments, the patient being treated according to the methods of the present disclosure has chronic kidney disease (CKD). In one embodiment, the patient has CKD anemia. In another embodiment, the patient has Kidney Disease Outcomes Quality Initiative (KDOQI) stages 1-5 CKD. In another embodiment, the patient has KDOQI stages 3-5 CKD. In another embodiment, the patient has an estimated glomerular filtration rate (eGFR) according to the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) of less than 60 mL / min / 1.73 m^2.

[0014] In some embodiments, the patient being treated according to the methods of the present disclosure has atherosclerotic cardiovascular disease (ASCVD), coronary artery disease (CAD), or peripheral artery disease (PAD).

[0015] In some embodiments, the patient has clonal hematopoiesis of indeterminate potential (CHIP). In some embodiments, the patient has one or more mutations in DNMT3A, TET2, ASXL1, PPM1D, TP53, JAK2, SF3B1, or SRSF2.

[0016] In some embodiments, the patient has a risk factor for developing cardiovascular disease. For example but not limited to, risk factors for developing cardiovascular disease are selected from the group consisting of: (a) diabetes (type I or type II); (b) metabolic syndrome; (c) hypertension; (d) male about 55 years of age or older or female about 65 years of age or older; (e) dyslipidemia; (f) family history of cardiovascular disease; (g) history of stroke or transient ischemic attack; (h) smoking (current smoking or history of past smoking); (i) hyperhomocysteinemia; (j) hyperuricemia; (k) male HDL-C level ≤ about 40 mg / dL or female HDL-C level ≤ about 50 mg / dL; (l) renal dysfunction (e.g., creatinine clearance rate (“CrCL”) greater than about 30 mL / min and less than about 60 mL / min); (m) retinopathy (e.g., non-proliferative retinopathy, pre-proliferative retinopathy, proliferative retinopathy, macularopathy, advanced diabetic eye disease, or history of photocoagulation); (n) microalbuminuria (e.g., microalbuminuria or other positive dipstick test, albumin / creatinine ratio ≥ about 2.5 mg / mmol, or albumin excretion rate ≥ about 20 mg / min at least two consecutive timed collections); (o) macroalbuminuria (e.g., Albustix or other dipstick shows proteinuria, albumin / creatinine ratio ≥ about 25 mg / mmol, or albumin excretion rate ≥ about 200 mg / min at least two consecutive timed collections); and / or (p) ankle-brachial index < about 0.9 without symptoms of intermittent claudication.

[0017] In some embodiments, the patient being treated according to the methods of the present disclosure has anemia of chronic inflammation, anemia of chronic disease, or iron-restricted anemia. In some embodiments, the patient has functional iron deficiency or iron restriction. In some embodiments, the patient has elevated hepcidin in serum or urine. In some embodiments, the patient has a red blood cell distribution width (RDW) > about 13% or the RDW is in the highest quartile of the general population. In some embodiments, the patient has a white blood cell count (WBC) > 9000 cells / μL of blood or the WBC is in the highest quartile of the general population.

[0018] In some embodiments, the patient had an infection in the past month, past three months, past six months, or past year. In some embodiments, the patient had a COVID-19 (SARS-CoV-2) infection in the past month, past three months, past six months, or past year. In some embodiments, the patient had surgery in the past month, past three months, past six months, or past year. In some embodiments, the patient has periodontal disease.

[0019] In some embodiments, the patient's absolute neutrophil count is not less than 2.0×10 per liter 9In some embodiments, the patient's platelet count is not less than 120×10 per liter. 9 In some embodiments, the patient's single urine to creatinine ratio is not less than 4.

[0020] In some embodiments, the patient is negative for active tuberculosis, HIV, or hepatitis B or C. In some embodiments, the patient has not received long-term immunosuppressive therapy.

[0021] In some embodiments, the cardiovascular disease is selected from the group consisting of non-fatal myocardial infarction, non-fatal stroke, and cardiovascular death. In one embodiment, the cardiovascular disease is heart failure.

[0022] In some embodiments, the therapeutically effective dose of the present disclosure is between about 5 mg and about 200 mg. In some embodiments, the therapeutically effective dose is about 5, about 7.5, about 10, about 15, about 20, about 25, about 30, about 50, about 60, about 70, about 80, about 90, or about 100 mg of an anti-IL-6 antibody or antibody fragment.

[0023] In some embodiments, the therapeutically effective dose is about 50 mg every 90 days. In one embodiment, the total therapeutically effective dose is about 100 mg. In some embodiments, the therapeutically effective dose is about 25 mg every 90 days. In one embodiment, the total therapeutically effective dose is about 50 mg. In some embodiments, the therapeutically effective dose is about 15 mg every 30 days. In one embodiment, the total therapeutically effective dose is about 90 mg.

[0024] In one embodiment, the therapeutically effective dose is administered subcutaneously.

[0025] In some embodiments, the dosing schedule of the anti-IL-6 antibody or antibody fragment is once every 1 week to once every 24 weeks. In one embodiment, the therapeutically effective dose is administered once every 4, 8, 12, or 24 weeks. In some embodiments, the therapeutically effective dose is administered once every 30 days to once every 90 days. In one embodiment, the therapeutically effective dose is administered once every 30 days. In one embodiment, the therapeutically effective dose is administered once every 90 days.

[0026] In some embodiments, the methods of the present disclosure include: (a) subcutaneously administering a loading dose of an anti-IL-6 antibody or antibody fragment to a patient at least for the first two doses during a loading regimen; and (b) thereafter subcutaneously administering a maintenance dose of the anti-IL-6 antibody or antibody fragment to the patient during a maintenance regimen. In some embodiments, the loading regimen includes administering the loading dose every 1 week, every 2 weeks, or every 4 weeks. In some embodiments, the maintenance regimen includes administering the maintenance dose every 4 weeks, every 8 weeks, every 12 weeks, or every 24 weeks. In one embodiment, the methods of the present disclosure include: (a) subcutaneously administering a loading dose of an anti-IL-6 antibody or antibody fragment to a patient every 4 weeks for the first two doses during a loading regimen; and (b) thereafter subcutaneously administering a maintenance dose of the anti-IL-6 antibody or antibody fragment to the patient every 8 or 12 weeks during a maintenance regimen. In some embodiments, the loading dose is greater than or equal to the maintenance dose. In some embodiments, the loading dose is between 5 mg and 200 mg. In some embodiments, the maintenance dose is between 5 mg and 200 mg.

[0027] In some embodiments, the patient has an inflammation. In one embodiment, the patient has an IL-6 mediated inflammation. In some embodiments, the treatment of the present disclosure is sufficient to reduce the inflammation without causing immunosuppression.

[0028] In some embodiments, immunosuppression is measured by an absolute neutrophil count (ANC). In some embodiments, the ANC after treatment is at least 500 cells / pL. In some embodiments, the ANC after treatment is at least 1000 cells / pL. In some embodiments, the ANC after treatment is at least 1500 cells / pL. In some embodiments, the ANC after treatment is at least 2000 cells / pL. In some embodiments, compared to the pre-treatment level, the decrease in ANC is no more than 2000 cells / pL. In some embodiments, compared to the pre-treatment level, the decrease in ANC is no more than 1500 cells / pL. In some embodiments, compared to the pre-treatment level, the decrease in ANC is no more than 1000 cells / pL. In some embodiments, compared to the pre-treatment level, the decrease in ANC is no more than 500 cells / pL. In some embodiments, compared to the pre-treatment level, the decrease in ANC is no more than about 50%. In some embodiments, compared to the pre-treatment level, the decrease in ANC is no more than about 40%. In some embodiments, compared to the pre-treatment level, the decrease in ANC is no more than about 30%. In some embodiments, compared to the pre-treatment level, the decrease in ANC is no more than about 20%. In some embodiments, compared to the pre-treatment level, the decrease in ANC is no more than about 10%. In some embodiments, compared to the pre-treatment level, the ANC has not decreased.

[0029] In some embodiments, inflammation is measured by high-sensitivity C-reactive protein (hsCRP) levels. In some embodiments, the pre-treatment hsCRP level of the patient is elevated. In some embodiments, the pre-treatment hsCRP level of the patient is at least 2 mg / L. In some embodiments, the pre-treatment hsCRP level of the patient is at least 4 mg / L. In some embodiments, the pre-treatment hsCRP level of the patient is at least 6 mg / L. In some embodiments, the pre-treatment hsCRP level of the patient is at least 10 mg / L. In some embodiments, the pre-treatment hsCRP level of the patient is 2 mg / L or lower. In some embodiments, the pre-treatment hsCRP level of the patient is 1 mg / L or lower. In some embodiments, the post-treatment hsCRP level does not exceed 2 mg / L. In some embodiments, the post-treatment hsCRP level does not exceed 1 mg / L. In some embodiments, the hsCRP level is reduced by at least about 50% compared to pre-treatment. In some embodiments, the hsCRP level is reduced by at least about 60% compared to the pre-treatment level. In some embodiments, the hsCRP level is reduced by at least about 70% compared to the pre-treatment level. In some embodiments, the hsCRP level is reduced by at least about 80% compared to the pre-treatment level. In some embodiments, the hsCRP level is reduced by at least about 90% compared to the pre-treatment level. In some embodiments, treatment results in a reduction of hsCRP within about 4, about 8, about 12, or about 24 weeks of treatment. In some embodiments, treatment results in a reduction of hsCRP after about 180 days of treatment. In some embodiments, treatment is sufficient to maintain the reduction of hsCRP for at least 24 or 48 weeks.

[0030] In some embodiments, treatment is sufficient to reduce fibrinogen, haptoglobin, serum amyloid A (SAA), secretory phospholipase A2 (sPLA2), lipoprotein(a), or neutrophil-to-lymphocyte ratio (NLR) by at least about 15%, about 30%, or about 50%.

[0031] In some embodiments, treatment is sufficient to reduce the incidence of one or more cardiovascular events by at least about 10%, about 15%, about 20%, about 25%, about 30%, about 40%, or about 50%. In some embodiments, adverse cardiovascular events are selected from the group consisting of death, cardiovascular death, non-fatal myocardial infarction, non-fatal stroke, unstable angina requiring urgent coronary revascularization, heart failure (new onset, worsening, or acute), ventricular arrhythmias due to ischemia, ischemic injury to cardiac valves, an increase in N-terminal pro-brain natriuretic peptide (NT-pro-BNP), an increase in cardiac injury markers (e.g., troponin or creatine kinase-MB), a decrease in left ventricular ejection fraction of at least about 5%, atrial fibrillation or other supraventricular arrhythmias, intestinal ischemia (small intestine or colon), new onset or worsening peripheral arterial disease, severe limb ischemia, atherosclerotic embolization (cholesterol embolization), new onset or worsening thromboangiitis obliterans, a worsening of New York Heart Association (NYHA) heart failure classification by one class or more, a worsening of the Kansas City Cardiomyopathy Questionnaire score, and a change in the Short Form-36 (SF-36) Physical Component Score (PCS).

[0032] In some embodiments, the methods described herein further comprise the step of treating the subject with an additional form of therapy. In some embodiments, the additional form of therapy comprises administering one or more therapeutic agents other than the anti-IL-6 antibody or antibody fragment described herein. Therapeutic agents include, but are not limited to, a second antibody (e.g., an anti-IL-1 antibody, an anti-IGF-1 receptor antibody, an anti-VEGF antibody, and / or an anti-IL17a antibody), a soluble receptor (e.g., a soluble IL-1 receptor, a soluble TNF-α receptor), an anti-inflammatory agent (e.g., paclitaxel, docetaxel, cisplatin, doxorubicin, prednisone, mitomycin, progesterone, tamoxifen, or fluorouracil), or a cardiovascular risk modulator (e.g., an antihypertensive drug, such as an adrenergic blocker, an angiotensin-converting enzyme inhibitor, an angiotensin II receptor blocker, and a calcium channel blocker; a lipid-lowering agent, such as a statin, a fibrate, a PCSK9 inhibitor, a bile acid resin, niacin, a selective cholesterol absorption inhibitor, an ω-3 fatty acid and fatty acid ester, and an adenosine triphosphate-citrate lyase (ACL) inhibitor; an anti-diabetic drug, such as metformin; or an antiplatelet drug, such as aspirin, clopidogrel, ticlopidine, ticagrelor, prasugrel, and cangrelor).

[0033] In some embodiments, an anti-IL-6 antibody or antibody fragment comprising the CDRs described herein is included in a pharmaceutical composition comprising the anti-IL-6 antibody or antibody fragment and a pharmaceutically acceptable carrier.

[0034] In some embodiments, there are provided pharmacological agents, compositions, methods, and / or dosing schedules that have certain advantages compared to currently used and / or agents, compositions, methods, and / or dosing schedules known in the art, including the ability to administer at a lower frequency or at a lower dose to achieve an equivalent effect of inhibiting IL-6-mediated signaling. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 Shows a schematic diagram of a Phase II randomized, double-blind, placebo-controlled trial of TOUR006. DETAILED DESCRIPTION

[0036] Provided herein are methods of treating cardiovascular disease, which comprise subcutaneously administering to a patient in need a therapeutically effective dose of an anti-interleukin-6 (anti-IL-6) antibody or antibody fragment.

[0037] Also provided herein are pharmacological agents, compositions, methods, and / or dosing schedules for treating cardiovascular disease.

[0038] Antibodies

[0039] Provided herein are antibodies that specifically bind IL-6 and antigen-binding fragments thereof. The antibodies and antigen-binding fragments disclosed herein specifically bind human IL-6. In some embodiments, the antibody may be specific only for human IL-6 and may not exhibit non-human cross-reactivity.

[0040] Throughout this disclosure, the term "about" may be used in connection with numerical values and / or ranges. The term "about" should be understood to cover values that are close to the stated value and within an acceptable error range in the art. For example, "about 40 [units]" may mean within a range of ±10%, ±9%, ±8%, ±7%, ±6%, ±5%, ±4%, ±3%, ±2%, ±1%, less than ±1%, or any other value or range of values therein or thereunder.

[0041] As used herein, the term "antibody" refers to an immunoglobulin (Ig) molecule and the immunologically active portion or fragment of an immunoglobulin molecule, i.e., a molecule containing an antigen-binding site that specifically binds to an antigen (e.g., IL-6) (reacting immunologically therewith). "Specifically binds" or "immunologically reacts" means that an antibody reacts with one or more antigenic determinants of the desired antigen and does not react with other polypeptides. In some embodiments, an antibody is said to specifically bind an antigen when it preferentially recognizes its target antigen in a complex mixture of proteins and / or macromolecules. In some embodiments, an antibody "specifically binds" IL-6 if it binds IL-6 with a higher affinity, higher avidity, more readily, and / or for a longer duration than it binds other polypeptides.

[0042] The term "antibody" broadly refers to an immunoglobulin (Ig) molecule, typically comprising four polypeptide chains: two heavy chains (H) and two light chains (L), or any functional fragment, mutant, variant, or derivative thereof that retains the basic target-binding characteristics of the Ig molecule. Such mutant, variant, or derivative antibody forms are known in the art.

[0043] In a full-length antibody, each heavy chain comprises a heavy chain variable region (abbreviated herein as the VH region) and a heavy chain constant region. The heavy chain constant region comprises three domains, CH1, CH2, and CH3. Each light chain comprises a light chain variable region (abbreviated herein as the VL region) and a light chain constant region. The light chain constant region comprises one domain, CL. The VH and VL domains can be further subdivided into hypervariable regions, called complementarity-determining regions (CDRs), interspersed with more conserved regions, called framework regions (FRs). Each VH domain and VL domain is composed 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.

[0044] The term "Fc region" is used to define the C-terminal region of an immunoglobulin heavy chain. The "Fc region" can be a native sequence Fc region or a variant Fc region. Although the boundaries of the Fc region of an immunoglobulin heavy chain may vary, the human IgG heavy chain Fc region is generally defined as the amino acid residues starting from Cys226 or extending from Pro230 to its carboxyl terminus. The numbering of residues in the Fc region is according to the EU numbering system. The Fc region of an immunoglobulin generally comprises two constant regions, CH2 and CH3. The Fc region can exist in a dimeric or monomeric form. The Fc region binds to various cell receptors (e.g., Fc receptors) and other immune molecules (e.g., complement proteins).

[0045] The immunoglobulin molecule can be of any type (e.g., IgG, IgE, IgM, IgD, IgA or IgY) and class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1 or IgA2) or subclass. IgG, IgD and IgE antibodies typically contain two identical heavy chains and two identical light chains as well as two antigen-binding domains, each composed of VH and VL. Typically, IgA antibodies are composed of two monomers, each monomer being composed of two heavy chains and two light chains (like IgG, IgD and IgE antibodies); thus, the IgA molecule has four antigen-binding domains, each domain being composed of one VH and one VL. Some IgA antibodies are monomeric as they are composed of two heavy chains and two light chains. Secreted IgM antibodies are generally composed of five monomers, each monomer being composed of two heavy chains and two light chains (as are IgG and IgE antibodies). Thus, the IgM molecule has ten antigen-binding domains, each domain being composed of one VH and one VL. The cell-surface form of IgM has a two heavy chain / two light chain structure similar to IgG, IgD and IgE antibodies.

[0046] The term "antigen-binding portion" or "antigen-binding fragment" (or "antibody portion" or "antibody fragment") of an antibody as used herein refers to one or more fragments of an antibody that retain the ability to specifically bind an antigen (e.g., IL-6). It has been demonstrated that the antigen-binding function of an antibody can be achieved by portions or fragments of a full-length antibody. Examples of binding fragments encompassed by the term "antigen-binding portion" of an antibody include (i) Fab fragments, monovalent fragments consisting of the VL, VH, CL, and CH1 domains; (ii) F(ab')2 fragments, divalent fragments containing two Fab fragments linked by a hinge region disulfide bond; (iii) Fd fragments consisting of the VH and CH1 domains; (iv) Fv fragments consisting of the VL and VH domains of a single arm of an antibody; (v) dAb (domain antibody) fragments (Ward et al., (1989) Nature 341:544-546; WO 90 / 05144A1, each of which is incorporated herein by reference in its entirety), which contain a single variable domain; and (vi) isolated complementarity-determining regions (CDRs). The present disclosure also encompasses Fab' fragments. Fab' fragments can be formed by reduction of F(ab')2 fragments. Fab' is derived from F(ab')2; thus, it may contain a small portion of the Fc. In addition, although the two domains VL and VH of an Fv fragment are encoded by different genes, they can be joined together using recombinant methods via a synthetic linker such that they can be made into a single protein chain in which the VL and VH domains pair to form a monovalent molecule (referred to as a single-chain Fv (scFv). See, e.g., Bird et al. (1988) Science 242:423-426; Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883. Such single-chain antibodies are also intended to be encompassed within the term "antigen-binding portion" of an antibody. In some embodiments, scFv molecules can be incorporated into fusion proteins. In some embodiments, single-chain camelid antibodies are provided herein. In some embodiments, shark heavy-chain antibodies (V-NAR) are provided herein. See English et al. (2020) Antibody Therapeutics, 3(1):1-9. Examples of antigen-binding portions are known in the art (Kontermann and Dubel, eds., Antibody Engineering (2001) Springer-Verlag. New York. p. 790). In some embodiments, single-domain antibodies are provided herein. Generally, the term "antibody" as used herein encompasses "antibody fragments". Antibody fragments generally retain the antigen-binding properties of the full-length antibody.

[0047] The antibodies and antibody portions provided herein can be in multispecific (e.g., bispecific or trispecific) forms. Such multispecific molecules specifically bind two or more different molecular targets or epitopes. In some embodiments, the antibody or antigen-binding portion is a bispecific molecule that specifically binds a first antigen and a second antigen, wherein the first antigen is IL-6 and the second antigen is not IL-6. In some embodiments, the antibody or antigen-binding portion is a diabody. A diabody is a bivalent bispecific antibody in which the VH and VL domains are expressed on a single polypeptide chain but using a linker that is too short to allow pairing between the two domains on the same chain, thus forcing the domains to pair with the complementary domains on the other chain and generating two antigen-binding sites (see, e.g., Holliger et al. (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448; Poljak et al. (1994) Structure 2:1121-1123). In some embodiments, the antibody or antigen-binding portion is a triabody, a tetrabody, a bis-scFv, or a tandem scFv. In some embodiments, the antibody or antigen-binding portion is a dual-affinity retargeting protein.

[0048] In some embodiments, the anti-IL-6 antigen-binding portion disclosed herein is a Fab, F(ab')2, Fab', Fv, scFv, Fd, single-domain antibody, single-chain camelid antibody, diabody, triabody, tetrabody, or bis-scFv.

[0049] As used herein, the terms “immunobinding” and “immunobinding properties” refer to non-covalent interactions that occur between an immunoglobulin molecule (e.g., an antibody or an antigen-binding portion thereof) and an antigen specific for the immunoglobulin. The strength or affinity of an immunobinding interaction can be expressed by the dissociation constant (K d ) of the interaction, where a smaller K d represents a greater affinity. The immunobinding properties of a selected polypeptide can be quantified using methods well known in the art. One such method involves measuring the rates of antigen-binding site / antigen complex formation and dissociation, where these rates depend on the concentration of the complex partners, the affinity of the interaction, and geometric parameters that have an equal effect on the rates in both directions. Thus, the “association rate constant” (K on ) and the “dissociation rate constant” (K off ) can both be determined by calculating the concentrations and the actual association and dissociation rates. (See Malmqvist, Nature 361:186-187 (1993)). The ratio of K off / K on cancels out all parameters unrelated to affinity and is equal to the dissociation constant K d. (See Davies et al. (1990) Annual Rev Biochem 59:439-473). When the equilibrium binding constant (K d ) is ≤ 10 μM, preferably ≤ 10 nM, more preferably ≤ 10 nM, and most preferably ≤ 100 pM to about 1 pM, the antibodies or antigen-binding portions provided herein are said to specifically bind IL-6, as measured by assays such as radioligand binding assays or similar assays known to those of skill in the art.

[0050] In some embodiments, the anti-IL-6 antibodies or antigen-binding portions provided herein are monovalent or divalent and comprise single-stranded or double-stranded. Functionally, the binding affinity of the antibody or antigen-binding portion can be in the range of about 10 -5 M to 10 -12 M. For example, the binding affinity of the antibody or antigen-binding portion is about 10 -6 M to 10 -12 M, about 10 -7 M to 10 -12 M, about 10 -8 M to 10 -12 M, about 10 -9 M to 10 -12 M, about 10 -5 M to 10 -11 M, about 10 -6 M to 10 -11 M, about 10 -7 M to 10 -11 M, about 10 -8 M to 10 - 11 M, about 10 -9 M to 10 -11 M, about 10 -10 M to 10 -11 M, about 10 -5 M to 10 -10 M, about 10 -6 M to 10 -10 M, about 10 -7 M to 10 -10 M, about 10 -8 M to 10 -10 M, about 10 -9 M to 10 -10 M, about 10 -5 M to 10 -9 M, about 10 -6 M to 10 -9 M, about 10 -7 M to 10 -9 M, about 10 -8 M to 10 -9M, about 10 -5 M to 10 -8 M, about 10 -6 M to 10 -8 M, about 10 -7 M to 10 -8 M, about 10 -5 M to 10 -7 M, about 10 -6 M to 10 -7 M or about 10 -5 M to 10 -6 M.

[0051] A human anti-IL-6 monoclonal antibody (PF-04236921) is described in US8,188,235 which is incorporated herein by reference in its entirety. The human anti-IL-6 monoclonal antibody is a fully human immunoglobulin G2 monoclonal antibody that binds to human IL-6 and has a half-life of 36 - 51 days. In phase I trials in healthy volunteers and patients with rheumatoid arthritis (protocol B0151001, NCT 00838565 and NCT01166555), intravenous and subcutaneous (SC) injection of the human anti-IL-6 monoclonal antibody (PF-04236921) was well tolerated and resulted in sustained inhibition of C-reactive protein (CRP), an inflammatory marker under the transcriptional control of IL-6. PF-04236921 was also tested in a phase II trial in patients with systemic lupus erythematosus (SLE; NCT01405196). Although the study did not reach the primary endpoint, the primary endpoint and key secondary endpoints were improved at 10 mg. Overall, the human anti-IL-6 monoclonal antibody demonstrated desirable pharmacokinetic (PK) and pharmacodynamic (PD) properties, supported sustained target inhibition, and had a low incidence of immunogenicity upon single and multiple dose administration. See Danese et al., Randomised trial and open-label extension study of an anti-interleukin-6 antibody in Crohn's disease (ANDANTE I and II), Gut 2019; 68:40–48; Li et al., Pharmacokinetics and C-reactive protein modelling of anti-interleukin-6 antibody (PF-04236921) in healthy volunteers and patients with autoimmune disease, Br J Clin Pharmacol. September 2018; 84(9):2059–2074.

[0052] Table 1 provides the amino acid and nucleic acid sequences of the human anti-IL-6 antibody (TOUR006).

[0053] Table 1. Amino acid and nucleic acid sequences of the human anti-IL-6 antibody

[0054]

[0055]

[0056] The sequences of CDR1, CDR2, and CDR3 (from left to right) are underlined in the heavy and light chains, respectively.

[0057] Provided herein is a method of treating cardiovascular disease, which comprises subcutaneously administering to a patient in need a therapeutically effective dose of an anti-interleukin-6 (anti-IL-6) antibody or antibody fragment having variable heavy chain (VH) CDRs defined in SEQ ID NOs: 2, 3, and 4, and variable light chain (VL) CDRs defined in SEQ ID NOs: 8, 9, and 10. In some embodiments, the antibody or antibody fragment comprises a heavy chain polypeptide and a light chain polypeptide, the heavy chain polypeptide comprising a polypeptide having at least about 95%, about 96%, about 97%, about 98%, or about 99% identity to SEQ ID NO: 1, and the light chain polypeptide comprising a polypeptide having at least about 95%, about 96%, about 97%, about 98%, or about 99% identity to SEQ ID NO: 7. In some embodiments, the antibody or antibody fragment comprises a heavy chain polypeptide and a light chain polypeptide, the heavy chain polypeptide comprising a polypeptide having the sequence of SEQ ID NO: 1, and the light chain polypeptide comprising a polypeptide having the sequence of SEQ ID NO: 7. In some embodiments, the anti-IL-6 antibody or antigen-binding portion comprises a human IgG2 constant region.

[0058] As used herein, the term "conservative substitution" refers to the replacement of one amino acid with another amino acid without significantly and detrimentally altering functional activity. A preferred example of a "conservative substitution" is the replacement of one amino acid with another amino acid having a value ≥0 in the following BLOSUM 62 substitution matrix (see Henikoff and Henikoff, 1992, PNAS 89:10915-10919):

[0059]

[0060] The calculation of sequence homology or identity between sequences (the terms are used interchangeably herein) can be performed as follows.

[0061] To determine the percent identity of two amino acid sequences or two nucleic acid sequences, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in one or both of the first and second amino acid or nucleic acid sequences for optimal alignment, and non-homologous sequences can be disregarded for comparison purposes). In a preferred embodiment, the length of the reference sequence aligned for comparison purposes is at least about 30%, preferably at least about 40%, more preferably at least about 50%, even more preferably at least about 60%, even more preferably at least about 70%, about 75%, about 80%, about 82%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% of the length of the reference sequence. The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, the molecules are identical at that position (as used herein, amino acid or nucleic acid “identity” is equivalent to amino acid or nucleic acid “homology”). The percent identity between two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps that need to be introduced to achieve optimal alignment of the two sequences and the length of each gap.

[0062] Sequence comparison and determination of the percent identity between two sequences can be accomplished using mathematical algorithms. In some embodiments, the percent identity between two amino acid sequences is determined as follows: using the Needleman et al. ((1970) J. Mol. Biol. 48:444 - 453) algorithm in the GAP program incorporated in the GCG software package, using a BLOSUM 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6 or 4 and a length weight of 1, 2, 3, 4, 5 or 6. In some embodiments, the percent identity between two nucleotide sequences is determined as follows: using the GAP program in the GCG software package, using the NWSgapdna.CMP matrix, and a gap weight of 40, 50, 60, 70 or 80 and a length weight of 1, 2, 3, 4, 5 or 6. A set of parameters (which can be used if the practitioner is unsure which parameters should be applied to determine whether a molecule is within the sequence identity or homology limits of the present invention) is the BLOSUM 62 scoring matrix, where the gap penalty is 12, the gap extension penalty is 4, and the frameshift gap penalty is 5.

[0063] In some embodiments, the percent identity between two amino acid or nucleotide sequences can be determined as follows: using the algorithm of Meyers et al. ((1989) CABIOS 4:11-17) incorporated into the ALIGN program (version 2.0), using a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4.

[0064] In some embodiments, the anti-IL-6 antibodies or antigen-binding portions provided herein are monoclonal.

[0065] In some embodiments, the anti-IL-6 antibodies or antigen-binding portions provided herein are chimeric. The term "chimeric" is intended to refer to an antibody molecule or an antigen-binding portion thereof, wherein the variable region sequence is derived from one species and at least one constant region sequence is derived from another species. For example, one or all of the variable domains of the light chain and / or one or all of the variable domains of the heavy chain of a murine antibody (e.g., a murine monoclonal antibody) can each be linked to a human constant region, such as but not limited to an IgG1, IgG2, or IgG4 human constant region. Examples of chimeric antibodies and suitable techniques for their production are provided in U.S. 4,816,567, U.S. 4,975,369, and U.S. 4,816,397, each of which is incorporated herein by reference in its entirety.

[0066] In some embodiments, the anti-IL-6 antibodies or antigen-binding portions provided herein are humanized. The term "humanized" is intended to refer to an antibody or an antigen-binding portion thereof that has been engineered to contain one or more human framework regions in the variable domain, as well as non-human (e.g., murine, rat, or hamster) CDRs of the heavy and / or light chain. In some embodiments, the humanized antibody comprises sequences that are entirely human except for the CDRs. In some embodiments, the VH domain, VL domain, or both the VH domain and VL domain of the anti-IL-6 antibodies or antigen-binding portions provided herein comprise one or more human framework region amino acid sequences. In some embodiments, the humanized antibody comprises sequences that are entirely human except for the CDRs, which are the CDRs of antibody 32G8H6. Examples of humanized antibodies and suitable techniques for their production are provided in: Hwang et al., Methods 36:35, 2005; Queen et al., Proc. Natl. Acad. Sci. USA, 86:10029-10033, 1989; Jones et al., Nature, 321:522-25, 1986; Riechmann et al., Nature, 332:323-27, 1988; Verhoeyen et al., Science, 239:1534-36, 1988; Orlandi et al., Proc. Natl. Acad. Sci. USA, 86:3833-37, 1989; U.S. 5,225,539; U.S. 5,530,101; U.S. 5,585,089; U.S. 5,693,761; U.S. 5,693,762; U.S. 6,180,370; and WO 90 / 07861, each of which is incorporated herein by reference in its entirety.

[0067] In some embodiments, humanization includes removing post-translational modification (PTM) sites in the variable domain sequences of the non-human antibody (e.g., in the CDR or framework sequences). For example, one or more PTM sites in the CDR sequence can be removed by replacing certain amino acid residues. In some embodiments, humanization includes CDR grafting and back mutation.

[0068] In some embodiments, the anti-IL-6 antibody or antigen-binding portion thereof comprises an immunoglobulin constant region. In some embodiments, the immunoglobulin constant region is IgG, IgE, IgM, IgD, IgA, or IgY. In some embodiments, the immunoglobulin constant region is IgG1, IgG2, IgG3, IgG4, IgA1, or IgA2. In some embodiments, the immunoglobulin constant region is immunologically inert. In some embodiments, the immunoglobulin constant region comprises one or more mutations to reduce or prevent FcγR binding, antibody-dependent cell-mediated cytotoxicity activity, and / or complement-dependent cytotoxicity activity. In some embodiments, the immunoglobulin constant region is a wild-type human IgG1 constant region; a wild-type human IgG2 constant region; a wild-type human IgG4 constant region; a human IgG1 constant region comprising the amino acid substitutions L234A, L235A, and G237A; a human IgG1 constant region comprising the amino acid substitutions L234A, L235A, G237A, and P331S; or a human IgG4 constant region comprising the amino acid substitution S228P; wherein the numbering is according to the EU numbering system. In some embodiments, the positions of amino acid residues in the constant region of the immunoglobulin molecule are numbered according to the EU nomenclature (Ward et al., 1995 Therap. Immunol. 2:77-94).

[0069] In some embodiments, the anti-IL-6 antibody or antigen-binding portion thereof may comprise an immunoglobulin light chain constant region, which is a κ light chain constant region or a λ light chain constant region.

[0070] In some embodiments, the anti-IL-6 antibody or antigen-binding portion thereof may comprise a human IgG4 constant region comprising the amino acid substitution S228P and a κ light chain constant region.

[0071] Also provided herein are immunoconjugates comprising an anti-IL-6 antibody or antigen-binding portion thereof linked to a therapeutic agent. In some embodiments, the therapeutic agent is a small molecule drug.

[0072] Drug composition

[0073] The anti-IL-6 antibodies and antigen-binding portions described herein (also referred to herein as "active compounds") can be incorporated into a pharmaceutical composition suitable for administration. Such compositions generally comprise an anti-IL-6 antibody or antigen-binding portion (or an immunoconjugate comprising the antibody or portion) and a pharmaceutically acceptable carrier, diluent, or excipient. As used herein, the term "pharmaceutically acceptable" refers to molecular entities and compositions that do not generally produce an allergic or other untoward reaction when administered by routes well known in the art. Molecular entities and compositions that are approved by a regulatory agency of the Federal or State government of the United States or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and particularly in humans, are considered "pharmaceutically acceptable." As used herein, the term "pharmaceutically acceptable carrier" is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are compatible with pharmaceutical administration. Suitable carriers are described in the most recent edition of the standard reference text Remington's Pharmaceutical Sciences, which is incorporated herein by reference. Some examples of such carriers or diluents include, but are not limited to, water, saline, Ringer's solution, dextrose solution, and 5% human serum albumin. Liposomes and non-aqueous vehicles, such as fixed oils, can also be used. The use of such media and agents for pharmaceutically active substances is well known in the art. Unless any conventional media or agent is incompatible with the active compound, it is contemplated that they will be used in the compositions. Supplementary active compounds can also be incorporated into the compositions.

[0074] Provided herein is a pharmaceutical composition comprising (i) an anti-IL-6 antibody or an antigen-binding portion thereof, wherein the antibody or antigen-binding portion comprises a VH domain and a VL domain, wherein: (a) the VH domain amino acid sequence comprises HCDR1 of SEQ ID NO:2, HCDR2 of SEQ ID NO:3, and HCDR3 of SEQ ID NO:4; and the VL domain amino acid sequence comprises LCDR1 of SEQ IDNO:8, LCDR2 of SEQ ID NO:9, and LCDR3 of SEQ ID NO:10; and (ii) a pharmaceutically acceptable carrier, diluent, or excipient.

[0075] The pharmaceutical compositions disclosed herein can be formulated to be compatible with their intended route of administration. Examples of routes of administration include parenteral administration, such as intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal (i.e., topical), transmucosal, and rectal administration. Solutions or suspensions for parenteral, intradermal, or subcutaneous application may include the following components: a sterile diluent, such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerin, propylene glycol, or other synthetic solvents; an antibacterial agent, such as benzyl alcohol or methylparaben; an antioxidant, such as ascorbic acid or sodium bisulfate; a chelating agent, such as ethylenediaminetetraacetic acid (EDTA); a buffer, such as acetate, citrate, or phosphate; and a reagent for adjusting tonicity, such as sodium chloride or glucose. Acids or bases (e.g., hydrochloric acid or sodium hydroxide) can be used to adjust the pH. Parenteral preparations may be enclosed in ampoules, disposable syringes, or multi-dose vials made of glass or plastic.

[0076] Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions (when soluble in water) or dispersions. For intravenous administration, suitable carriers include saline, bacteriostatic water, (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). In all cases, the composition must be sterile and should be a fluid that is easy to inject. It must be stable under the conditions of preparation and storage and must be preserved against the action of microorganisms such as bacteria and fungi. The carrier can be a solvent or a dispersion medium containing, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, etc.) and suitable mixtures thereof. For example, by using a coating such as lecithin, by maintaining the required particle size in the case of a dispersion, and by using surfactants, the appropriate fluidity can be maintained. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents (e.g., parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, etc.). In many cases, it will be preferable to include in the composition an isotonic agent, such as sugars, polyols (such as mannitol, sorbitol), sodium chloride. Prolonged absorption of injectable compositions can be achieved by including a reagent for prolonging absorption, such as aluminum monostearate and gelatin, in the composition.

[0077] Sterile injectable solutions can be prepared by incorporating the required amount of the compound, with one or a combination of the above-listed active ingredients, into a suitable solvent as required, followed by filtration sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle which contains a basic dispersion medium and the required other ingredients listed above. In the case of sterile powders for the preparation of sterile injectable solutions, the methods of preparation are vacuum drying and freeze drying, which yield a powder of the active ingredient and any additional desired ingredients from their previously sterile filtered solutions.

[0078] Oral compositions generally include an inert diluent or an edible carrier. They can be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the active compounds can be mixed with excipients and used in the form of tablets, lozenges, or capsules. A fluid carrier can also be used to prepare oral compositions for use as mouthwashes, where the compounds in the fluid carrier are taken orally, swished around, and either spat out or swallowed. Binders and / or adjuvant materials of pharmaceutical compatibility can be included as part of the composition. Tablets, pills, capsules, lozenges, etc. can contain any of the following ingredients or compounds of similar nature: binders such as microcrystalline cellulose, tragacanth, or gelatin; excipients such as starch or lactose; disintegrants such as alginic acid, or corn starch; lubricants such as magnesium stearate; glidants such as colloidal silicon dioxide; sweetening agents such as sucrose or saccharin; or flavoring agents such as peppermint, methyl salicylate, or orange flavoring.

[0079] For inhaled administration, the compounds can be delivered in the form of an aerosol spray from a pressure vessel or dispenser containing a suitable propellant (such as a gas like carbon dioxide, etc.) or a nebulizer.

[0080] Systemic administration can also be effected by transmucosal or transdermal means. For transmucosal or transdermal administration, a penetrant suitable for the barrier to be permeated is used in the formulation. Such penetrants are well known in the art and include, for example, detergents, bile salts, and fusidic acid derivatives for transmucosal administration. Transmucosal administration can be achieved by using nasal sprays or suppositories. For transdermal administration, the active compounds are formulated into ointments, pastes, gels, or creams well known in the art.

[0081] The pharmaceutical agent can also be made in the form of suppositories (e.g., using conventional suppository bases such as cocoa butter and other glycerides) or retention enemas for rectal delivery.

[0082] In some embodiments, the active compounds are prepared with a carrier that will protect the compounds from rapid elimination by the body, such as a controlled-release formulation, including implants and microcapsule delivery systems. Biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid can be used. Those skilled in the art will be clear about the preparation methods of such formulations. Such materials are also commercially available. Liposome suspensions can also be used as pharmaceutically acceptable carriers.

[0083] It is particularly advantageous to formulate oral or parenteral compositions in dosage unit form for ease of administration and uniform dosage. As used herein, dosage unit form refers to physically discrete units suitable as a single dose for a subject to be treated; each unit contains a predetermined quantity of the active compound calculated to produce the desired therapeutic effect in association with the desired pharmaceutical carrier. The specifications of the dosage unit forms of the invention are dictated by and directly dependent on the unique characteristics of the active compound and the particular therapeutic effect to be achieved, and the inherent limitations of the art of synthesizing such active compounds for the treatment of individuals.

[0084] The pharmaceutical compositions provided herein may be contained in a container, package, or dispenser together with instructions for administration.

[0085] Use of the antibody

[0086] Provided herein are methods and uses of the anti-IL-6 antibodies, anti-IL-6 antigen-binding portions, immunoconjugates, and pharmaceutical compositions described herein for providing a therapeutic benefit to a subject having a disorder associated with IL-6 expression. In some embodiments, the disorder is an elevated cardiovascular risk. In some embodiments, the disorder is chronic kidney disease (CKD) with an elevated cardiovascular risk. In some embodiments, the disorder is a cardiovascular disease, including but not limited to non-fatal myocardial infarction, non-fatal stroke, cardiovascular death, and heart failure.

[0087] In some embodiments, the methods described herein further comprise the step of treating the subject with an additional form of therapy. In some embodiments, the additional form of therapy includes administering one or more therapeutic agents other than the anti-IL-6 antibody or antibody fragment described herein. Therapeutic agents include but are not limited to a second antibody (e.g., an anti-IL-1 antibody, an anti-IGF-1 receptor antibody, an anti-VEGF antibody, and / or an anti-IL17a antibody), a soluble receptor (e.g., a soluble IL-1 receptor, a soluble TNF-α receptor), an anti-inflammatory agent (e.g., paclitaxel, docetaxel, cisplatin, doxorubicin, prednisone, mitomycin, progesterone, tamoxifen, or fluorouracil), or a cardiovascular risk modulator (e.g., an antihypertensive drug, such as an adrenergic blocker, an angiotensin-converting enzyme inhibitor, an angiotensin II receptor blocker, and a calcium channel blocker; a lipid-lowering agent, such as a statin, a fibrate, a PCSK9 inhibitor, a bile acid resin, niacin, a selective cholesterol absorption inhibitor, an ω-3 fatty acid and fatty acid ester, and an adenosine triphosphate-citrate lyase (ACL) inhibitor; an anti-diabetic drug, such as metformin; or an antiplatelet drug, such as aspirin, clopidogrel, ticlopidine, ticagrelor, prasugrel, and cangrelor).

[0088] Provided herein is the anti-IL-6 antibody or anti-IL-6 antigen-binding portion, immunoconjugate, or pharmaceutical composition described herein for use as a medicament.

[0089] As used herein, the term "effective amount" or "therapeutically effective amount" refers to an amount of an agent (e.g., an anti-IL-6 antibody or an antigen-binding portion thereof) that is sufficient to reduce or ameliorate a disorder (e.g., a cardiovascular disease) or the severity and / or duration of one or more of its symptoms, prevent the progression of the disease, cause regression of the disease, prevent the recurrence, development, onset or progression of one or more symptoms associated with the disorder, detect the disorder, or enhance or improve the prophylactic or therapeutic effect of another therapy (e.g., a prophylactic or therapeutic agent). In some embodiments, the therapeutically effective dose of the anti-IL-6 antibody or antibody fragment effectively alters one or more biomarkers of IL-6-mediated signal transduction, including but not limited to total sIL-6R, total IL-6, C-reactive protein (CRP), and / or autoantibodies, for a surprisingly extended period of time.

[0090] As used herein, the terms "treat", "treating", "treatment", etc. refer to alleviating or ameliorating a dz and / or its associated signs or symptoms, or slowing or halting its progression. It should be understood that treating a disorder or condition does not necessarily require complete elimination of the disorder, condition or symptoms associated therewith, although this is not excluded.

[0091] As used herein, "pre-treatment" means prior to the first administration of an anti-IL-6 antibody according to the methods described herein. Pre-treatment does not exclude and typically includes prior administration of therapies other than an anti-IL-6 antibody.

[0092] As used herein, "post-treatment" means after the administration of an anti-IL-6 antibody according to the methods described herein. Post-treatment includes after administration of any anti-IL-6 antibody at any dose described herein. Post-treatment also includes after the anti-IL-6 antibody treatment phase.

[0093] It should be noted that there are some unclear abbreviations like "bz" and "dz" in the original text which might need further clarification in the source context for more accurate translation.The actual dosage, administration rate, and administration schedule will depend on the nature and severity of the disease being treated, the specific mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the delivery site of the composition, the method of administration, the dosing schedule, and other factors known to the medical practitioner. Treatment prescriptions (such as dose determination, etc.) are the responsibility of general practitioners and other physicians and may depend on the severity and / or progression of the symptoms of the disease being treated. Appropriate dosages of antibody molecules are well known in the art (Ledermann J.A. et al., 1991, Int. J. Cancer 47:659-664; Bagshawe K.D. et al., 1991, Antibody, Immuno conjugates and Radiopharmaceuticals 4:915-922). Specific dosages may be noted herein or in the Physician’s Desk Reference (2003) as being appropriate for the type of drug being administered. The therapeutically effective amount or appropriate dosage of an antibody molecule can be determined by comparing its in vitro activity and in vivo activity in animal models. Methods for extrapolating effective dosages in mice and other test animals to humans are known. The exact dosage will depend on a variety of factors, including whether the antibody is used for prophylaxis or treatment, the size and location of the area to be treated, the exact nature of the antibody (e.g., whole antibody, fragment), and the nature of any detectable label or other molecule attached to the antibody.

[0094] Typical antibody dosages range from 100 μg to 1 g for systemic application and from 1 μg to 1 mg for intradermal injection. An initial higher loading dose may be administered, followed by one or more lower doses. In some embodiments, the antibody is a whole antibody, such as an IgG1, IgG2, or IgG4 isotype. This is the dosage for a single treatment of an adult subject, which can be adjusted proportionally for children and infants, and the dosage of other antibody forms can also be adjusted according to the molecular weight ratio. Depending on the doctor's judgment, the treatment can be repeated at daily, twice-weekly, weekly, or monthly intervals. The treatment schedule for the subject may depend on the pharmacokinetic and pharmacodynamic properties of the antibody composition, the route of administration, and the nature of the disorder being treated. In some embodiments, the administration of the present disclosure comprises an amount of an anti-IL-6 antibody or antibody fragment of at least about 10 mg, or at least about 20 mg, or at least about 30 mg, or at least about 40 mg, or at least about 50 mg.

[0095] Treatment can be periodic, and the time interval between administrations can be about two weeks or longer, such as about three weeks or longer, about four weeks or longer, about once a month or longer, about five weeks or longer, or about six weeks or longer. For example, treatment can be administered every two to four weeks or every four to eight weeks. Treatment can be carried out before and / or after surgery, and / or can be directly administered or applied at the anatomical site of surgical treatment or invasive surgery. Suitable formulations and routes of administration are as described above. In some embodiments, the dosing schedule of the anti-IL-6 antibody or antibody fragment is once every 4 or 8 weeks, for a total of up to about 52 weeks.

[0096] In some embodiments, the subject is a human, non-human primate, pig, horse, cow, dog, cat, guinea pig, mouse, or rat. In some embodiments, the subject is an adult. In some embodiments, the subject is a child. In some embodiments, the patient has received or is receiving prednisone, etc. at the start of the treatment. In one aspect, the dose of prednisone, etc. is 10 mg to 60 mg per day.

[0097] The section headings used herein are for organizational purposes only and should not be construed as limiting the subject matter. All documents or portions of documents cited herein, including but not limited to patents, patent applications, articles, books, and papers, are hereby expressly incorporated by reference in their entirety for any purpose. If the definitions of terms defined in one or more of the incorporated documents or portions of documents conflict with the definitions of such terms in the present application, the definitions in the present application shall control. However, any references, articles, publications, patents, patent publications, and patent applications cited herein do not constitute and should not be regarded as admitting or implying in any way that they constitute valid prior art or a part of the common general knowledge in any country of the world.

[0098] In this specification, unless otherwise specified, any concentration range, percentage range, ratio range, or integer range should be understood to include any integer value within the range, and, where appropriate, its fractions (such as one-tenth and one-hundredth of an integer). It should be understood that, unless otherwise specified, the terms "a" and "an" as used herein refer to "one or more" of the recited components. The use of alternatives (e.g., "or") should be understood to mean one, both, or any combination of the alternatives. As used herein, the terms "comprising" and "including" are synonyms.

[0099] The present disclosure will be further illustrated by the following examples, which are intended to be merely illustrative of the present disclosure and in no way limiting.

[0100] Examples

[0101] Example 1: Clinical Evaluation of Human Anti-IL6 Antibody in Patients with Cardiovascular Disease

[0102] The Phase II clinical study of the human anti-IL6 antibody TOUR006 disclosed in this article is to be conducted in patients at high risk of cardiovascular events, including chronic kidney disease (CKD). The study is a randomized, double-blind, placebo-controlled trial designed to evaluate the anti-inflammatory effects of three dosing regimens of TOUR006 on participants with elevated cardiovascular risk (hsCRP ≥ 2.0 mg / L at baseline and diagnosed with CKD).

[0103] Inclusion criteria

[0104] Participants must meet all of the following criteria to participate in the study:

[0105] 1) ≥ 18 years of age at the time of signing the informed consent form (ICF).

[0106] 2) Serum hsCRP level ≥ 2.0 mg / L at the screening visit.

[0107] 3) Diagnosed with CKD (estimated glomerular filtration rate [eGFR] ≥ 15 and < 60 mL / min / 1.73m 2 ) and not expected to require dialysis during the study.

[0108] 4) Vaccinated against COVID-19 (completed the primary series) at least 30 days before the screening visit and / or previously infected with COVID-19 (except for COVID-19 infection within 30 days before the screening visit [see exclusion criterion #19]).

[0109] 5) Agree to comply with contraceptive and fertility restrictions:

[0110] a. Male participants must have undergone surgical sterilization or, if having sexual intercourse with a female partner of childbearing age, must also agree to use condoms during the study and for 4 months after completion of the study drug administration.

[0111] b. Female participants of childbearing potential (including women who started menopause less than 2 years before the screening visit, had non-treatment-induced amenorrhea less than 12 months before the screening visit, or had not undergone surgical sterilization [no ovaries and / or uterus and / or two fallopian tubes]) must have a negative serum pregnancy test at the screening visit, negative urine pregnancy tests at all time points specified in the protocol, and agree to use at least 1 acceptable contraceptive method throughout the study and for 32 weeks after the last study drug administration.

[0112] Exclusion criteria

[0113] Participants will be excluded from the study if they meet any of the following criteria:

[0114] Laboratory values

[0115] 1) Hemoglobin value ≤ 8.0 g / dL at screening visit.

[0116] 2) Transferrin saturation < 15.0% at screening visit.

[0117] 3) Absolute neutrophil count < 2.0×10 9 / L at screening visit.

[0118] 4) Platelet count < 120×10 9 / L at screening visit.

[0119] 5) Single urine protein to creatinine ratio > 3000 mg / g (3.0 g / g) at screening visit.

[0120] 6) Serum albumin < 3.0 g / dL at screening visit.

[0121] 7) Serum total IgG < 700 mg / dL at screening visit.

[0122] 8) Uncontrolled diabetes with HbA1c > 9% at screening visit.

[0123] 9) Fasting low-density lipoprotein > 130 mg / dL at screening visit.

[0124] 10) Fasting triglycerides > 200 mg / dL at screening visit.

[0125] 11) Alanine aminotransferase (ALT) or aspartate aminotransferase (AST) > 2.5 times the upper limit of normal at screening visit.

[0126] 12) Positive tuberculosis test at screening visit. A blood test (e.g., QuantiFERON) is preferred, but a purified protein derivative (PPD) skin test read by a qualified healthcare professional within 48 - 72 hours may also be performed. If a participant has a positive PPD test but a negative QuantiFERON test, the participant is eligible to participate in the study.

[0127] 13) Evidence of human immunodeficiency virus (HIV)-1 or HIV-2 infection detected by serological testing at screening visit.

[0128] 14) Evidence of hepatitis B or hepatitis C detected by serological examination at screening visit (e.g., positive hepatitis B surface antigen or hepatitis C antibody).

[0129] 15) Serum vitamin B-12 and folate levels below the laboratory normal lower limit at screening visit (for anemia causes other than known or suspected CKD).

[0130] Medical condition or disease

[0131] 16) Had an acute coronary syndrome, ischemic stroke, transient ischemic attack, or other thromboembolic event within 6 months prior to randomization.

[0132] 17) Participants with an indwelling urinary catheter or intravenous catheter.

[0133] 18) Had clinical evidence of or suspected active infection.

[0134] 19) Had a history of severe gastrointestinal bleeding (excluding hemorrhoid bleeding) within 6 months prior to randomization.

[0135] 20) Had a history of active diverticulitis or gastrointestinal abscess within 12 months prior to randomization.

[0136] 21) Had a clinically active history of inflammatory bowel disease within 12 months prior to randomization.

[0137] 22) Uncontrolled hypertension (defined as mean systolic blood pressure > 160 mmHg or mean diastolic blood pressure > 100 mmHg in three measurements prior to randomization). If antihypertensive treatment was initiated or increased due to initial screening blood pressure being above these limits, the investigator may, at their discretion, re-evaluate the participant according to this criterion.

[0138] 23) Planned coronary revascularization (percutaneous coronary intervention or coronary artery bypass grafting) or any other major surgical procedure during the course of the study.

[0139] 24) Had a history of major cardiac surgery, non-cardiac surgery, or major endoscopic surgery within 6 months prior to randomization.

[0140] 25) New York Heart Association class IV congestive heart failure.

[0141] 26) Had a history of hospitalization for worsening heart failure within 3 months prior to randomization.

[0142] 27) Untreated arrhythmias, including:

[0143] a. Hemodynamically significant arrhythmias

[0144] b. Atrial fibrillation or atrial flutter with uncontrolled resting heart rate (> 100 bpm)

[0145] c. Any other arrhythmia determined by the investigator that may affect the safety of the participant or confound the interpretation of safety data in this study.

[0146] 28) QTc > 450 milliseconds in males or QTc > 470 milliseconds in females

[0147] 29) Diagnosed with cachexia and / or unexpected weight loss > 5% within 3 months before randomization.

[0148] 30) Diagnosed with protein-losing enteropathy.

[0149] 31) Received active treatment or had active malignancy within 12 months before randomization.

[0150] 32) History of bone marrow or solid organ transplantation, or expected to receive bone marrow or solid organ transplantation during the study participation.

[0151] 33) Immunodeficiency (hereditary or acquired, such as acquired immunodeficiency syndrome, common variable immunodeficiency, etc.).

[0152] 34) Had a severe infection within 12 months before screening (requiring hospitalization and / or intravenous [IV] antibiotics, IV antifungal or IV antiviral treatment and / or an infection with clinical manifestations considered by the investigator to meet the criteria for severe infection) or had more than 1 such infection within 36 months before randomization.

[0153] 35) COVID-19 infection (including mild or asymptomatic) that is ongoing or occurred within 30 days before randomization.

[0154] 36) Any history of severe opportunistic infection (previous or current) (e.g., non-localized thrush due to corticosteroid treatment).

[0155] 37) History of systemic lupus erythematosus or clinical suspicion of systemic lupus erythematosus.

[0156] 38) Any live attenuated vaccine administration planned during the course of this study.

[0157] 39) Known allergy to the study drug or any of its components.

[0158] 40) Any history of substance use disorder, including alcohol abuse.

[0159] 41) History of seizures within 10 years before randomization.

[0160] 42) Any history of mental illness.

[0161] 43) Severe cognitive impairment.

[0162] Previous or current medications

[0163] 44) Received the study drug within 30 days before randomization (or 5 half-lives of the administered study drug, whichever is longer).

[0164] 45) Received any immunomodulators or immunosuppressive agents, including systemic corticosteroids, within 5 drug half-lives (or 3 months, whichever is longer) prior to randomization, or expected to use such drugs at any time during the study. Note: The use of ear, ophthalmic, inhaled, and topical corticosteroids or local corticosteroid injections is not excluded.

[0165] 46) Received hypoxia-inducing factor stabilizers (e.g., molidustat, Roxadustat) or erythropoietin analogs (e.g., erythropoietin α or β, darbopoietin α, or continuous erythropoietin receptor activator) within 6 weeks prior to randomization.

[0166] 47) Expected to receive any study drug or any excluded drug specified in the protocol during the treatment period or safety follow-up period.

[0167] 48) Used systemic antibiotics, antiviral drugs, or antifungal drugs within 14 days prior to randomization ("systemic" defined as oral or IV drugs absorbed into the bloodstream).

[0168] General Exclusions

[0169] 49) Currently breastfeeding.

[0170] 50) Expected to require blood transfusions during the course of the study.

[0171] 51) Any condition that may interfere with the conduct of the study or the interpretation of the study results, or whose treatment may interfere with the conduct of the study or the interpretation of the study results, or that in the opinion of the investigator would increase the risk of participating in the study.

[0172] 52) Life expectancy < 2 years.

[0173] The main inclusion criteria include chronic kidney disease (CKD), and a pre-treatment hsCRP level ≥ 2 mg / L. For example but not limited to, eligible patients include those with a pre-treatment hsCRP level ≥ 2 mg / L, CKD patients, CKD anemia patients, patients with stages 1 - 5 CKD according to the Kidney Disease Outcomes Quality Initiative (KDOQI), patients with an estimated glomerular filtration rate (eGFR) below 60 mL / min / 1.73 m^2 according to the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI), and / or patients with clonal hematopoiesis of indeterminate potential (CHIP) features. CHIP features include, but are not limited to, one or more mutations in DNMT3A, TET2, ASXL1, PPM1D, TP53, JAK2, SF3B1, or SRSF2 found in blood or bone marrow cells.

[0174] The study also included patients with a confirmed cardiovascular disease (CVD). For example, but not limited to, eligible patients included those with atherosclerotic cardiovascular disease (ASCVD), coronary artery disease (CAD), or peripheral artery disease (PAD).

[0175] In addition, the study also encompassed patients with risk factors for developing cardiovascular disease, whether or not they had CKD. For example, but not limited to, risk factors for cardiovascular disease were selected from the group consisting of: (a) diabetes (type I or type II); (b) metabolic syndrome; (c) hypertension; (d) males approximately 55 years of age or older or females approximately 65 years of age or older; (e) dyslipidemia; (f) family history of cardiovascular disease; (g) history of stroke or transient ischemic attack; (h) smoking (current or past smoking history); (i) hyperhomocysteinemia; (j) hyperuricemia; (k) male HDL-C level ≤ approximately 40 mg / dL or female HDL-C level ≤ approximately 50 mg / dL; (l) renal dysfunction (e.g., creatinine clearance rate (“CrCL”) greater than approximately 30 mL / min and less than approximately 60 mL / min); (m) retinopathy (e.g., non-proliferative retinopathy, pre-proliferative retinopathy, proliferative retinopathy, macularopathy, advanced diabetic eye disease, or history of photocoagulation); (n) microalbuminuria (e.g., microalbuminuria or other positive dipstick test, albumin / creatinine ratio ≥ approximately 2.5 mg / mmol, or albumin excretion rate ≥ approximately 20 mg / min in at least two consecutive timed collections); (o) macroalbuminuria (e.g., Albustix or other dipstick shows macroalbuminuria, albumin / creatinine ratio ≥ approximately 25 mg / mmol, or albumin excretion rate ≥ approximately 200 mg / min in at least two consecutive timed collections); and (p) ankle-brachial index < approximately 0.9 without symptoms of intermittent claudication.

[0176] The study may also have included patients with: chronic inflammatory anemia, anemia of chronic disease, or iron-restricted anemia; functional iron deficiency or iron restriction; elevated hepcidin (serum or urine); red blood cell distribution width (RDW) of approximately > 13% or RDW in the highest quartile of the general population; and / or white blood cell count (WBC) of > 9000 cells per microliter of blood or WBC in the highest quartile of the general population.

[0177] The study may also have included patients who had been infected with a virus in the past month, past 3 months, past 6 months, or past year; had been infected with COVID-19 (SARS-CoV-2) in the past month, past 3 months, past 6 months, or past year; had undergone surgery in the past month, past 3 months, past 6 months, or past year; and / or had periodontal disease.

[0178] The study may also include patients with an absolute neutrophil count of at least 2.0×10 9 / L; a platelet count of at least 120×10 9 / L; and a single urine-to-creatinine ratio of at least 4.

[0179] The study excluded those who were detected with active tuberculosis, HIV, or hepatitis B or C. The study also excluded those who were on long-term immunosuppressive therapy.

[0180] Dose regimen

[0181] The study protocol is as Figure 1 shown. Eligible patients will be randomly assigned in a 1:1:1:1 ratio to receive one of three dosing regimens of TOUR006 or placebo (see treatment groups below). To mitigate imbalances in background safety risks of CKD, participants will be stratified by CKD stage to ensure balanced randomization. The treatment groups are as follows:

[0182] · Treatment Group A (administer 50 mg of TOUR006 subcutaneously every 90 days)

[0183] ○ Participants will receive 50 mg TOUR006 injections on Day 1 and Day 90 of the study; they will receive corresponding placebo injections on Days 30, 60, 120, and 150 of the study.

[0184] ○ Cumulative total dose of TOUR006: 100 mg

[0185] · Treatment Group B (administer 25 mg of TOUR006 subcutaneously every 90 days)

[0186] ○ Participants will receive 25 mg TOUR006 injections on Day 1 and Day 90 of the study; they will receive corresponding placebo injections on Days 30, 60, 120, and 150 of the study.

[0187] ○ Cumulative total dose of TOUR006: 50 mg

[0188] · Treatment Group C (administer 15 mg of TOUR006 subcutaneously every 30 days)

[0189] ○ Participants will receive 15 mg TOUR006 injections on Days 1, 30, 60, 90, 120, and 150 of the study.

[0190] ○ Cumulative total dose of TOUR006: 90 mg

[0191] · Placebo: administered subcutaneously once every 30 days

[0192] Approximately 120 participants (30 in each treatment group) will be randomly assigned. After the screening period ends, the randomized participants will receive TOUR006 or placebo every 30 days (participants in groups A and B will receive placebo on study days 30, 60, 120, and 150 to comply with the 30-day dosing schedule). We will follow the participants for pharmacokinetic and pharmacodynamic endpoints until study day 180 and for safety until study day 365. The primary pharmacodynamic endpoint assessment will be conducted on study day 180.

[0193] Other TOUR006 dosing regimens include three dose levels in the range of 5 mg to 200 mg. For example, the effective doses are 5, 7.5, 10, 15, 20, 25, 30, or 50 mg of the TOUR006 antibody. TOUR006 will be administered subcutaneously once every 4, 8, 12, or 24 weeks. The total study time for an individual patient is approximately 24 weeks.

[0194] Further dosing regimens include: (a) during the loading regimen, at least the first two doses are administered subcutaneously to the patient as a loading dose of TOUR006; and (b) thereafter, during the maintenance regimen, a maintenance dose of TOUR006 is administered subcutaneously to the patient. The loading regimen includes administering the loading dose once every 1 week, once every 2 weeks, or once every 4 weeks. The maintenance regimen includes administering the maintenance dose once every 4 weeks, once every 8 weeks, once every 12 weeks, or once every 24 weeks. The loading dose can be greater than or equal to the maintenance dose. The loading dose is 5 mg to 200 mg, and the maintenance dose is 5 mg to 200 mg.

[0195] Endpoint

[0196] Clinical relevant parameters will be recorded during the study, including but not limited to high-sensitivity C-reactive protein (hsCRP), absolute neutrophil count (ANC), fibrinogen, haptoglobin, serum amyloid A (SAA), secretory phospholipase A2 (sPLA2), lipoprotein(a) level, neutrophil-to-lymphocyte ratio (NLR), red blood cell distribution width (RDW), LDL level, APOB, APOA1, hemoglobin, transferrin saturation (TSAT), reticulocyte hemoglobin content (Chr), total iron-binding capacity (TIBC), serum ferritin, albumin, erythropoietic resistance index (ERI), grip strength, NT-proBNP, and cardiac MRI. Details of these clinical relevant parameters are described in US8,906,964, US11,369,582, and US11,384,143, the contents of each of which are hereby incorporated by reference in their entirety for any purpose.

[0197] The primary endpoint of this study was the change in hsCRP relative to baseline after 180 days of treatment. Additionally, the primary endpoint was the reduction in hsCRP at weeks 12 to 24 of treatment, such as at week 12, week 16, or week 24 of treatment. The enrolled patients would have inflammation or IL6-mediated inflammation. C-reactive protein (CRP) is a marker of inflammation. CRP levels increase with inflammation and can be measured by the hsCRP (high-sensitivity C-reactive protein) test. The hsCRP of patients before treatment was usually greater than 2 mg / L. In some cases, the hsCRP level of patients before treatment was 1 mg / L or lower. The treatment was sufficient to reduce the hsCRP level. For example, the hsCRP level after treatment did not exceed 2 mg / L or 1 mg / L. Compared with the pre-treatment level (baseline), the treatment was sufficient to reduce the hsCRP level by at least about 50%, about 60%, about 70%, about 80%, or about 90%. The treatment would result in a reduction in hsCRP within 4, 8, 12, 16, or 24 weeks of treatment. Additionally, the treatment was sufficient to maintain the reduction in hsCRP for at least 24 or 48 weeks.

[0198] One secondary endpoint was the proportion of participants who achieved an hsCRP response (a decrease >90% from baseline and / or hsCRP <2.0 mg / L) after 180 days of treatment. Another secondary endpoint was the serum drug concentration of TOUR006 at baseline and 30, 60, 90, 120, 150, and 180 days after treatment, as well as at 210, 270, 330, and 365 days after treatment.

[0199] To evaluate the safety and tolerability of TOUR006 in participants with elevated cardiovascular risk and CKD, the safety endpoints included the proportion of participants who experienced adverse events (AEs), serious AEs (SAEs), severe AEs, and AEs leading to discontinuation of treatment; the description and frequency of events of particular interest by treatment group; and the description of additional safety assessments by treatment group and dose, such as vital signs, electrocardiograms, and anti-drug antibodies.

[0200] To explore the effect of TOUR006 on inflammation markers, the exploratory endpoints included the change in serum amyloid A (SAA) relative to baseline; the change in lipoprotein(a) relative to baseline; the change in neutrophil-to-lymphocyte ratio (NLR) relative to baseline; and the change in secretory phospholipase (sPLA2) relative to baseline. To evaluate the effect of TOUR006 on functional iron deficiency and anemia, additional endpoints included the change in red blood cell distribution width (RDW) relative to baseline; the change in hemoglobin relative to baseline; and the change in transferrin saturation (TSAT), reticulocyte hemoglobin content (Chr), total iron-binding capacity (TIBC), and serum ferritin relative to baseline.

[0201] The treatment is sufficient to reduce inflammation without causing immunosuppression. Immunosuppression is measured by the absolute neutrophil count (ANC). The normal range of ANC is from about 500 to 7000 cells / pL; the post-treatment ANC is at least 500, 1000, 1500, or 2000 cells / pL compared to the pre-treatment level. In some cases, the decrease in ANC does not exceed about 50%, about 40%, about 30%, about 20%, or about 10% compared to the pre-treatment level. In some cases, the ANC does not decrease compared to the pre-treatment level.

[0202] Other secondary endpoints of this study include the incidence of one or more cardiovascular events. The treatment will reduce the incidence of one or more cardiovascular events by at least about 10%, about 15%, about 20%, about 25%, about 30%, about 40%, or about 50%. Adverse cardiovascular events include but are not limited to death, cardiovascular death, non-fatal myocardial infarction, non-fatal stroke, unstable angina requiring urgent coronary revascularization, heart failure (new onset, worsening, or acute), ventricular arrhythmia due to ischemia, ischemic injury of cardiac valves, increase in N-terminal pro-brain natriuretic peptide (NT-pro-BNP), increase in cardiac injury markers (such as troponin or creatine kinase-MB), decrease in left ventricular ejection fraction by at least about 5%, atrial fibrillation or other supraventricular arrhythmias, intestinal ischemia (small intestine or colon), new onset or worsening of peripheral arterial disease, severe limb ischemia, atherosclerotic embolism (cholesterol embolism), new onset or worsening of thromboangiitis obliterans, deterioration of New York Heart Association (NYHA) heart failure classification by one level or more, worsening of Kansas City Cardiomyopathy Questionnaire score, and change in the Short Form 36 (SF-36) Physical Component Score (PCS).

[0203] The study consists of three phases: (i) screening and randomization, (ii) treatment, and (iii) follow-up. During the screening process, each potential subject will provide informed consent before starting any specific procedure of the study. Subjects will be randomly assigned to the study groups centrally using a randomization scheme reviewed and approved by independent statisticians through an Interactive Web Response System (IWRS). During the treatment period, the randomly assigned subjects will receive treatment and evaluation according to the protocol. Follow-up will be conducted at 8 weeks, 12 weeks, 16 weeks, 24 weeks, or 36 weeks after the end of treatment.

[0204] Incorporated by reference

[0205] All references, articles, publications, patents, patent publications, and patent applications cited herein are incorporated by reference in their entirety for all purposes. However, any reference, article, publication, patent, patent publication, and patent application cited herein does not constitute and should not be construed as an admission or imply in any way that it constitutes valid prior art or a part of the common general knowledge in any country of the world.

Claims

1. A method for treating cardiovascular diseases, which comprises administering to a patient in need thereof a therapeutically effective dose of an anti-interleukin-6 (anti-IL-6) antibody or antibody fragment, said antibody or antibody fragment having variable heavy chain (VH) CDRs defined in SEQ ID NOs 2, 3, and 4, and variable light chain (VL) CDRs defined in SEQ ID NOs 8, 9, and 10.

2. The method according to claim 1, wherein said antibody or antibody fragment comprises a heavy chain polypeptide and a light chain polypeptide, said heavy chain polypeptide comprising a polypeptide having at least 98% identity with SEQ ID NO:1, and said light chain polypeptide comprising a polypeptide having at least 98% identity with SEQ ID NO:

7.

3. The method according to claim 1, wherein said antibody or antibody fragment comprises a heavy chain polypeptide having the sequence of SEQ ID NO:1 and a light chain polypeptide having the sequence of SEQ ID NO:

7.

4. The method according to any one of claims 1 to 3, wherein the patient is at least 18 years old.

5. The method according to any one of claims 1 to 4, wherein the patient has chronic kidney disease (CKD).

6. The method according to any one of claims 1 to 4, wherein the patient has CKD anemia.

7. The method according to claim 5 or 6, wherein the patient has Kidney Disease Outcomes Quality Initiative (KDOQI) stages 3 - 5 CKD.

8. The method according to claim 5 or 6, wherein the patient's estimated glomerular filtration rate (eGFR) Chronic Kidney Disease - Epidemiology Collaboration (CKD - EPI) is less than 60 mL / min / 1.73 m^2.

9. The method according to any one of claims 1 to 4, wherein the patient has atherosclerotic cardiovascular disease (ASCVD), coronary artery disease (CAD), or peripheral artery disease (PAD).

10. The method according to any one of claims 1 to 9, wherein the patient has a risk factor for developing cardiovascular disease.

11. The method according to claim 10, wherein the risk factor for developing cardiovascular disease is selected from the group consisting of: (a) type I or type II diabetes; (b) metabolic syndrome; (c) hypertension; (d) male approximately 55 years old or older or female approximately 65 years old or older; (e) dyslipidemia; (f) family history of cardiovascular disease; (g) history of stroke or transient ischemic attack; (h) current or past smoking history; (i) hyperhomocysteinemia; (j) hyperuricemia; (k) male HDL - C level ≤ approximately 40 mg / dL or female HDL - C level ≤ approximately 50 mg / dL; (l) renal dysfunction; (m) retinopathy; (n) microalbuminuria; (o) macroalbuminuria; and (p) ankle - brachial index < approximately 0.9 without symptoms of intermittent claudication.

12. The method according to any one of claims 1 to 11, wherein the patient has chronic inflammatory anemia, anemia of chronic disease, or iron - restricted anemia.

13. The method according to any one of claims 1 to 12, wherein the patient has functional iron deficiency or iron restriction.

14. The method according to any one of claims 1 to 13, wherein the hepcidin in the serum or urine of the patient is elevated.

15. The method according to any one of claims 1 to 14, wherein the red blood cell distribution width (RDW) of the patient is greater than 13% or the RDW is in the highest quartile.

16. The method according to any one of claims 1 to 15, wherein the white blood cell count (WBC) of the patient is more than 9000 cells per microliter of blood or the WBC is in the highest quartile.

17. The method according to any one of claims 1 to 16, wherein the absolute neutrophil count of the patient is not less than 2.0×10 9 / L.

18. The method according to any one of claims 1 to 17, wherein the platelet count of the patient is not less than 120×10 9 / L.

19. The method according to any one of claims 1 to 18, wherein the ratio of a single urine sample to creatinine of the patient is not less than 4.

20. The method according to any one of claims 1 to 19, wherein the patient is negative for active tuberculosis.

21. The method according to any one of claims 1 to 20, wherein the patient is negative for HIV.

22. The method according to any one of claims 1 to 21, wherein the patient is negative for hepatitis B or hepatitis C.

23. The method according to any one of claims 1 to 22, wherein the cardiovascular disease is selected from the group consisting of non-fatal myocardial infarction, non-fatal stroke, and cardiovascular death.

24. The method according to any one of claims 1 to 22, wherein the cardiovascular disease is heart failure.

25. The method according to any one of claims 1 to 24, wherein the therapeutically effective dose is about 5 mg to about 200 mg.

26. The method according to claim 25, wherein the therapeutically effective dose is about 5, about 7.5, about 10, about 15, about 20, about 25, about 30, about 50, about 60, about 70, about 80, about 90, or about 100 mg.

27. The method according to claim 26, wherein the therapeutically effective dose is about 50 mg every 90 days.

28. The method according to claim 27, wherein the total therapeutically effective dose is about 100 mg.

29. The method according to claim 26, wherein the therapeutically effective dose is about 25 mg every 90 days.

30. The method according to claim 29, wherein the total therapeutically effective dose is about 50 mg.

31. The method according to claim 26, wherein the therapeutically effective dose is about 15 mg every 30 days.

32. The method according to claim 31, wherein the total therapeutically effective dose is about 90 mg.

33. The method according to any one of claims 1 to 32, wherein the therapeutically effective dose is administered subcutaneously.

34. The method according to any one of claims 1 to 26, wherein the therapeutically effective dose is administered once every 1 week to every 24 weeks.

35. The method according to claim 26, wherein the therapeutically effective dose is administered once every 4 weeks, 8 weeks, 12 weeks, or 24 weeks.

36. The method according to any one of claims 1 to 26, wherein the therapeutically effective dose is administered once every 30 days to every 90 days.

37. The method according to claim 36, wherein the therapeutically effective dose is administered once every 30 days.

38. The method according to claim 36, wherein the therapeutically effective dose is administered once every 90 days.

39. The method according to any one of claims 1 to 26, further comprising: a) administering a loading dose of the anti-IL-6 antibody or antibody fragment to the patient in at least the first two doses during a loading regimen; and b) subsequently administering a maintenance dose of the anti-IL-6 antibody or antibody fragment to the patient during a maintenance regimen.

40. The method according to claim 39, wherein the loading regimen comprises administering the loading dose once every 1 week, once every 2 weeks, or once every 4 weeks.

41. The method according to claim 39, wherein the maintenance regimen comprises administering the maintenance dose once every 4 weeks, once every 8 weeks, once every 12 weeks, or once every 24 weeks.

42. The method according to claim 39, further comprising: (a) administering a loading dose of the anti-IL-6 antibody or antibody fragment to the patient once every 4 weeks in the first two doses during a loading regimen; and (b) subsequently administering a maintenance dose of the anti-IL-6 antibody or antibody fragment to the patient once every 8 or 12 weeks during a maintenance regimen.

43. The method according to any one of claims 39 to 42, wherein the loading dose is greater than or equal to the maintenance dose.

44. The method according to any one of claims 1 to 43, wherein the patient has an inflammation.

45. The method according to claim 44, wherein the patient has an IL-6-mediated inflammation.

46. The method according to claim 44 or 45, wherein the treatment is sufficient to reduce the inflammation without causing immunosuppression.

47. The method according to claim 44, wherein the immunosuppression is measured by an absolute neutrophil count (ANC).

48. The method according to claim 47, wherein the ANC after treatment is at least 500, 1000, 1500, or 2000 cells / pL.

49. The method according to claim 47, wherein the reduction of the ANC after treatment compared to the pre-treatment level is no more than 50%, 40%, 30%, 20%, or 10%.

50. The method according to claim 47, wherein the ANC does not decrease compared to the pre-treatment level.

51. The method according to any one of claims 44 to 50, wherein the inflammation is measured by a high-sensitivity C-reactive protein (hsCRP) level.

52. The method according to claim 51, wherein the pre-treatment hsCRP level of the patient is higher than 2 mg / L.

53. The method according to claim 51, wherein the pre-treatment hsCRP level of the patient is 2 mg / L or lower.

54. The method according to claim 51, wherein the pre-treatment hsCRP level of the patient is 1 mg / L or lower.

55. The method according to claim 51, wherein the treatment is sufficient to reduce the hsCRP level to 2 mg / L or lower.

56. The method according to claim 51, wherein the treatment is sufficient to reduce the hsCRP level to 1 mg / L or less.

57. The method according to any one of claims 51 to 56, wherein the treatment is sufficient to reduce the hsCRP level by at least 50%, 60%, 70%, 80% or 90% compared to the pre-treatment level.

58. The method according to claim 57, wherein the treatment causes a reduction in hsCRP within 4, 8 or 12 weeks of treatment.

59. The method according to claim 57, wherein the treatment is sufficient to maintain the reduction in hsCRP for at least 24 or 48 weeks.

60. The method according to claim 55 or 57, wherein the treatment causes a reduction in hsCRP after approximately 180 days of treatment.

61. The method according to any one of claims 1 to 60, wherein the treatment is sufficient to reduce fibrinogen, haptoglobin, serum amyloid A (SAA), secretory phospholipase A2 (sPLA2), lipoprotein(a) or neutrophil-to-lymphocyte ratio (NLR) by at least 15%, 30% or 50%.

62. The method according to any one of claims 1 to 61, wherein the treatment is sufficient to reduce the incidence of one or more cardiovascular events by at least 10%, 15%, 20%, 25%, 30%, 40% or 50%.

63. The method according to claim 62, wherein the adverse cardiovascular events are selected from the group consisting of: death, cardiovascular death, non-fatal myocardial infarction, non-fatal stroke, unstable angina requiring urgent coronary revascularization, new-onset heart failure, worsening heart failure, acute heart failure, ventricular arrhythmia due to ischemia, ischemic injury to cardiac valves, increase in N-terminal pro-brain natriuretic peptide (NT-pro-BNP), increase in cardiac injury markers, reduction in left ventricular ejection fraction by at least 5%, atrial fibrillation, supraventricular arrhythmia, small intestinal ischemia, colonic ischemia, new-onset or worsening peripheral arterial disease, severe limb ischemia, atherosclerotic embolism, cholesterol embolism, new-onset or worsening thromboangiitis obliterans, worsening of New York Heart Association (NYHA) heart failure classification by one or more grades, worsening of Kansas City Cardiomyopathy Questionnaire score, and change in Short Form 36 (SF-36) Physical Component Score (PCS).

64. The method according to claim 63, wherein the cardiac injury marker is troponin or creatine kinase-MB.

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