Methods of treating metabolic disorders with FGF21 variants

Abstract

Provided herein are methods of treating, preventing and managing metabolic disorders or cardiovascular disorders, and methods of reducing cardiovascular risk with FGF21 protein variants, including Fc-FGF21 variant fusion proteins.

Description

[0001] This application is a divisional application of Chinese patent application 201880086404.3. The filing date of the original application is December 21, 2018, and the name of the invention is “Method for treating metabolic disorders with FGF21 variants”.

[0002] I. Technical field

[0003] The present disclosure relates to methods of treating, preventing and managing metabolic disorders or cardiovascular disorders, and to methods of reducing cardiovascular risk using FGF21 protein variants, including Fc-FGF21 variant fusion proteins.

[0004] II. Background Technology

[0005] Insulin resistance is common and plays an important role in the pathogenesis of various diseases. Central adiposity is an important source of inflammatory cytokines and non-esterified fatty acids (which have been shown to have adverse effects on insulin sensitivity). Metabolic disorders associated with insulin resistance and adiposity manifest in various forms, such as diabetes, obesity, dyslipidemia, coronary heart disease and NAFLD / NASH. Fibroblast growth factor 21 (FGF21) is believed to act on the liver and adipose tissue to improve insulin sensitivity, reduce triglycerides and reduce adiposity. Therefore, FGF21 analogs with clinically acceptable dosing regimens are expected to become cutting-edge therapies for these metabolic disorders.

[0006] FGF21 is a member of the fibroblast growth factor family, and has a variety of metabolic functions. FGF21 is mainly released from hepatocytes, and to a lesser extent from fat and pancreatic beta cells. In mice, FGF21 has been shown to be the key regulator of metabolism (wherein induced expression under the conditions of both hunger and obesity). FGF21 induces various beneficial metabolic effects, including weight loss in non-clinical models and suffering from type 2 diabetes (T2DM) patients, improves insulin sensitivity, increases high-density lipoprotein-cholesterol (HDL-C), and reduces low-density lipoprotein-cholesterol (LDL-C), triglycerides (TG) and liver TG content. In addition, in non-clinical diabetic animal models, FGF21 effectively reduces glucose and does not induce hypoglycemia. In T2DM patients with increased body mass index, treatment with an FGF21 protein analog for four weeks resulted in significant improvements in lipid profiles, suggesting the translatability of the metabolic effects observed in animal models to humans (Gaich et al., 2013, Cell Metab. 18(3):333-40; Dong et al., 2015, Br J Clin Pharmacol. doi:10.1111 / bcp.12676). Gaich et al. described LY2405319 (a variant of FGF21) in a randomized, placebo-controlled, double-blind proof-of-concept trial in patients with obesity and type 2 diabetes, in which patients received placebo or 3 mg, 10 mg, or 20 mg of LY2405319 daily for 28 days. Dong et al. described a first-in-human study to evaluate the pharmacokinetics / pharmacodynamics (PK / PD), safety, and tolerability of a single intravenous (IV) dose of PF-05231023, a long-acting fibroblast growth factor 21 (FGF21) analog, and reported that PF-05231023 levels peaked immediately after IV administration, with mean terminal half-lives of 6.5 h-7.7 h and 66.5 h-96.6 h for the intact C- and N-termini, respectively.

[0007] There remains a need for improved FGF21 therapeutics with clinically acceptable dosing regimens for treating metabolic diseases and reducing cardiovascular risk in patients.

[0008] III. Summary of the Invention

[0009] Novel modifications to FGF21 can improve half-life and / or potency compared to wild-type FGF21 to provide FGF21 therapeutics for treating or managing metabolic or cardiovascular disorders (e.g., alleviating one or more symptoms of the disorder) or for reducing cardiovascular risk with clinically preferred dosing regimens.

[0010] In a particular aspect, provided herein are FGF21 protein variants (e.g., FGF21 protein variants described in Table 1, such as Fc-fusion proteins) and pharmaceutical compositions comprising such FGF21 protein variants for use in methods of treating, preventing and / or managing hypertriglyceridemia and heart disease risk, insulin resistance (e.g., patients with genetic mutations of the insulin receptor) and lipodystrophy, diabetes, obesity and non-alcoholic fatty liver disease (NAFLD) / non-alcoholic steatohepatitis (NASH).

[0011] V103 is a genetically engineered variant of human FGF21 that is stabilized by the introduction of a new disulfide bond and fused to human IgG1 Fc at its N-terminus (see PCT Publication WO 2013 / 049247, which is hereby incorporated by reference in its entirety). The combination of stability and fusion of the human FGF21 mutant with IgG1 Fc results in an approximately 50-100-fold increase in the half-life of cynomolgus monkeys (5 to 8 days) compared to wild-type (WT) human FGF21 (about 2 hours).

[0012] V103 repeats the effects disclosed by FGF21 in the insulin-resistant T2DM animal model: including the reduction of body weight, glucose, insulin, serum TG and liver TG content. V103 also shows a significant reduction in liver lipids, fibrosis and inflammation in a non-alcoholic fatty liver disease (NASH) mouse model. Therefore, provided herein is a pharmaceutical composition comprising FGF21 protein variant V103 (SEQ ID NO: 11) for the treatment, prevention and / or management of metabolic disorders (including hypertriglyceridemia and heart disease risk, insulin resistance (e.g., patients with genetic mutations of insulin receptors) and lipodystrophy, diabetes, obesity and non-alcoholic fatty liver disease (NAFLD) / non-alcoholic fatty liver disease (NASH).

[0013] In one aspect, provided herein is a human FGF21 protein variant for use in a method for treating, preventing or managing a metabolic disorder or cardiovascular disorder of a human subject, wherein the human FGF21 protein variant is provided for administration with a dosage in the range of 100 mg to 600 mg. In one aspect, provided herein is a human FGF21 protein variant for use in a method for treating, preventing or managing a metabolic disorder or cardiovascular disorder of a human subject, wherein the human FGF21 protein variant is provided for administration with an amount in the range of 100 mg to 600 mg.

[0014] In some embodiments of this aspect, the metabolic disorder or cardiovascular disorder is selected from hypercholesterolemia, dyslipidemia, hypertriglyceridemia, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), type 2 diabetes, and obesity.

[0015] In some embodiments of this aspect, the treatment, prevention or management of metabolic disorders or cardiovascular disorders comprises or is characterized by reducing one or more of the following: the subject's weight, liver fat content, elevated LDL-C, total C, triglycerides and Apo B levels. In certain embodiments thereof, the treatment, prevention or management of metabolic disorders or cardiovascular disorders comprises or is characterized by increasing the subject's HDL-C levels. In some embodiments thereof, the treatment, prevention or management of metabolic disorders or cardiovascular disorders comprises or is characterized by reducing the subject's triglyceride levels by at least about 40% or at least about 50%. In some embodiments thereof, the treatment, prevention or management of metabolic disorders or cardiovascular disorders comprises or is characterized by reducing the subject's cardiovascular risk.

[0016] In some embodiments of this aspect, the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, wherein the mature human FGF21 protein or a fragment thereof comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G and G202A numbered according to SEQ ID NO: 1. In certain embodiments thereof, the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11.

[0017] In some embodiments of this aspect, human FGF21 protein variants are provided for administration at a dosage of at least 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, or 400 mg. In some embodiments thereof, human FGF21 protein variants are provided for administration at a dosage of about 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, or 190 mg, optionally wherein the human FGF21 protein variants are provided at a dosage of about 100 mg or 150 mg. In some embodiments thereof, the human FGF21 protein variant is provided for administration at a dose of approximately 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg, optionally wherein the human FGF21 protein variant is provided at a dose of approximately 200 mg, 250 mg, or 300 mg.

[0018] In some embodiments of this aspect, human FGF21 protein variants are provided in the form of subcutaneous administration. In some embodiments, human FGF21 protein variants are provided for administration once a month or once every 4 weeks, once every 3 weeks, once every 2 weeks, or once a week. In some embodiments thereof, human FGF21 protein variants are provided for administration once a month or once every 4 weeks, once every 3 weeks, once every 2 weeks, or once a week. In some embodiments thereof, human FGF21 protein variants are provided in the form of subcutaneous administration.

[0019] In one aspect, provided herein is a human FGF21 protein variant in the manufacture of a medicine for treating, preventing or managing a metabolic disorder or cardiovascular disorder, wherein the unit dose of the human FGF21 protein variant is in the range of 100 mg to 600 mg. In one aspect, provided herein is a human FGF21 protein variant in the manufacture of a medicine for treating, preventing or managing a metabolic disorder or cardiovascular disorder, wherein the amount of the human FGF21 protein variant is in the range of 100 mg to 600 mg.

[0020] In some embodiments of this aspect, the metabolic disorder or cardiovascular disorder is selected from hypercholesterolemia, dyslipidemia, hypertriglyceridemia, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), type 2 diabetes, and obesity.

[0021] In some embodiments of this aspect, the treatment, prevention or management of metabolic disorders or cardiovascular disorders comprises or is characterized by reducing one or more of the following: body weight, liver fat content, elevated LDL-C, total C, triglycerides and Apo B levels. In some embodiments thereof, the treatment, prevention or management of metabolic disorders or cardiovascular disorders comprises or is characterized by increasing HDL-C levels. In some embodiments thereof, triglyceride levels are reduced by at least about 40% or at least about 50%. In some embodiments thereof, the treatment, prevention or management of metabolic disorders or cardiovascular disorders comprises or is characterized by reducing cardiovascular risk.

[0022] In some embodiments of this aspect, the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, wherein the mature human FGF21 protein or a fragment thereof comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G and G202A numbered according to SEQ ID NO: 1. In some embodiments thereof, the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11.

[0023] In some embodiments of this aspect, human FGF21 protein variants are provided for use with a dosage of at least 100mg, 150mg, 200mg, 250mg, 300mg, 350mg or 400mg. In certain embodiments, human FGF21 protein variants are provided for use with a dosage of about 100mg, 150mg, 200mg, 250mg, 300mg, 350mg or 400mg. In certain embodiments, human FGF21 protein variants are provided for use with a dosage of about 100mg, 110mg, 120mg, 130mg, 140mg, 150mg, 160mg, 170mg, 180mg or 190mg, optionally wherein the human FGF21 protein variants are provided for use with a dosage of about 100mg or 150mg. In some embodiments, the human FGF21 protein variant is provided for administration at a dose of approximately 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg, optionally wherein the human FGF21 protein variant is provided for administration at a dose of approximately 200 mg, 250 mg or 300 mg.

[0024] In some embodiments, human FGF21 protein variants are provided for administration once a month or once every 4 weeks, once every 3 weeks, once every 2 weeks, or once a week. In some embodiments, human FGF21 protein variants are provided for administration in the form of once a month or once every 4 weeks, once every 3 weeks, once every 2 weeks, or once a week. In some embodiments, human FGF21 protein variants are formulated for subcutaneous administration. In some embodiments, human FGF21 protein variants are provided in the form of subcutaneous administration.

[0025] In one aspect, provided herein is a method for treating, preventing or managing a metabolic disorder or cardiovascular disorder in a human subject, wherein the method includes administering human FGF21 protein variants to a subject at a dosage in the range of 100 mg to 600 mg. In one aspect, provided herein is a method for treating, preventing or managing a metabolic disorder or cardiovascular disorder in a human subject, wherein the method includes administering human FGF21 protein variants to a subject at an amount in the range of 100 mg to 600 mg.

[0026] In some embodiments, the metabolic disorder or cardiovascular disorder is selected from hypercholesterolemia, dyslipidemia, hypertriglyceridemia, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), type 2 diabetes, and obesity.

[0027] In some embodiments, the treatment, prevention or management of metabolic disorders or cardiovascular disorders comprises or is characterized by reducing one or more of the following: the subject's weight, liver fat content, elevated LDL-C, total C, triglycerides and Apo B levels. In certain embodiments, the treatment, prevention or management of metabolic disorders or cardiovascular disorders comprises or is characterized by increasing the subject's HDL-C levels. In some embodiments, the treatment, prevention or management of metabolic disorders or cardiovascular disorders comprises or is characterized by reducing the subject's triglyceride levels by at least about 40% or at least about 50%. In some embodiments, the treatment, prevention or management of metabolic disorders or cardiovascular disorders comprises or is characterized by reducing the subject's cardiovascular risk.

[0028] In some embodiments, the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, wherein the mature human FGF21 protein or a fragment thereof comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G and G202A numbered according to SEQ ID NO: 1. In certain embodiments, the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11.

[0029] In some embodiments, the method includes administering human FGF21 protein variants to a subject at a dosage of at least 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, or 400 mg. In some embodiments, the method includes administering human FGF21 protein variants to a subject at a dosage of about 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, or 190 mg. In some embodiments, the method includes administering human FGF21 protein variants to a subject at a dosage of about 100 mg or 150 mg. In some embodiments, the method comprises administering human FGF21 protein variants to a subject at a dosage of about 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg. In some embodiments, the method comprises administering human FGF21 protein variants to a subject at a dosage of about 200 mg, 250 mg, or 300 mg.

[0030] In some embodiments, human FGF21 protein variants are administered once a month or once every 4 weeks, once every 3 weeks, once every 2 weeks, or once a week. In some embodiments, human FGF21 protein variants are administered once a month or once every 4 weeks, once every 3 weeks, once every 2 weeks, or once a week. In some embodiments, the method includes administering human FGF21 protein variants in a form suitable for subcutaneous administration. In some embodiments, the method includes subcutaneous administration of human FGF21 protein variants. In some embodiments, human FGF21 protein variants are administered subcutaneously.

[0031] Each of the foregoing aspects and embodiments, as well as other elements described herein, may be combined in any manner and without limitation.

[0032] IV. Specific Implementation Methods

[0033] Provided herein are methods and pharmaceutical compositions for use in treating, preventing or managing metabolic disorders or cardiovascular disorders (e.g., alleviating one or more symptoms of the disorder), comprising administering a human FGF21 protein variant (e.g., Fc-FGF21 mutant fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose or amount in the range of 100 mg to 600 mg. In a particular aspect, the human FGF21 protein variant (e.g., V103 (SEQ ID NO: 11)) is administered at a dose in the range of 200 mg to 400 mg. In a particular aspect, the human FGF21 protein variant (e.g., V103 (SEQ ID NO: 11)) is administered at a dose of at least 200 mg, at least 300 mg, or at least 400 mg. In a specific aspect, the human FGF21 protein variant (e.g., an Fc-FGF21 mutant fusion protein, such as V103 (SEQ ID NO: 11)) is administered at a dose ranging from 100 mg to 600 mg (e.g., 250 mg to 350 mg) once every 4 weeks (or once a month), or once every 3 weeks or once every 2 weeks.

[0034] In one aspect, non-limiting examples of metabolic or cardiovascular disorders to be treated, prevented or managed by the methods provided herein include hypertriglyceridemia, diabetes (e.g., type 2 diabetes), obesity, type 1 diabetes, pancreatitis, dyslipidemia, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), insulin resistance, hyperinsulinemia, glucose intolerance, hyperglycemia, metabolic syndrome, hypertension, cardiovascular disease, acute myocardial infarction, atherosclerosis, peripheral arterial disease, stroke, heart failure, coronary heart disease, kidney disease, diabetic complications, neuropathy, diseases associated with severe inactivating mutations in the insulin receptor, and / or gastroparesis.

[0035] the term

[0036] With respect to FGF21, the term "native" or "wild type" refers to biologically active naturally occurring FGF21, including biologically active naturally occurring FGF21 variants. An exemplary human FGF21 wild type sequence has NCBI reference sequence number NP061986.1 and can be found in such issued patents as US6,716,626B1 (SEQ ID NO: 1).

[0037]

[0038] The corresponding mRNA sequence encoding the full-length FGF21 polypeptide (NCBI reference sequence number NM 019113.2) is shown below (SEQ ID NO: 2):

[0039]

[0040]

[0041] The mature FGF21 sequence lacks a leader sequence and may also include other modifications of the polypeptide, such as proteolytic processing of the amino terminus (with or without a leader sequence) and / or the carboxyl terminus, cleavage of smaller polypeptides from larger precursors, N-linked and / or O-linked glycosylation, and other post-translational modifications understood by those skilled in the art. A representative example of a mature FGF21 sequence has the following sequence (SEQ ID NO: 3, which represents amino acid positions 29-209 of the full-length FGF21 protein sequence (NCBI reference sequence number NP061986.1)):

[0042]

[0043] The corresponding cDNA sequence encoding the mature FGF21 polypeptide (SEQ ID NO: 3) is shown below (SEQ ID NO: 4):

[0044]

[0045]

[0046] The terms "FGF21 protein variant", "human FGF21 variant", "FGF21 polypeptide or protein variant", "FGF21 mutant" or any similar terms are defined as comprising human FGF21 in which the naturally occurring (i.e., wild-type) FGF21 amino acid sequence has been modified (e.g., wherein at least one amino acid of the wild-type protein has been substituted and / or removed by another amino acid). Additionally, the variant may include N- and / or C-terminal truncations relative to the wild-type FGF21 protein, or internal amino acid deletions relative to the wild-type human FGF21 protein. In general, the variant has some modified properties, structures or functions of the wild-type protein. For example, the variant may have enhanced or improved physical stability in concentrated solutions (e.g., less hydrophobic-mediated aggregation), enhanced or improved plasma stability when incubated with plasma, and enhanced or improved biological activity while maintaining a favorable spectrum of biological activities.

[0047] The acceptable amino acid substitutions and modifications constituting the differences between the FGF21 polypeptides and protein variants and mutants and wild-type FGF21 of the methods provided herein include, but are not limited to, one or more amino acid substitutions (including substitutions with non-naturally occurring amino acid analogs) and truncations. Therefore, FGF21 protein variants include, but are not limited to, site-directed FGF21 mutants, truncated FGF21 polypeptides, proteolytically resistant FGF21 mutants, reduced aggregated FGF21 mutants, FGF21 combination mutants, FGF21 conjugates (e.g., fatty acid-FGF21 conjugates, PEG-FGF21 conjugates) and FGF21 fusion proteins (e.g., Fc domain fusion proteins, human serum albumin fusion proteins) as described herein.

[0048] Variants may have increased compatibility with pharmaceutical preservatives (e.g., metacresol, phenol, benzyl alcohol), so that pharmaceutical formulations that maintain the physiochemical properties of proteins and the preservation of biological activity during storage can be prepared. Therefore, variants with enhanced drug stability relative to wild-type FGF21 have improved physical stability in concentrated solutions under physiological and preserved pharmaceutical formulation conditions while maintaining biological efficacy. As a set of non-limiting examples, variants provided herein may be more resistant to proteolysis and enzymatic degradation; may have improved stability; and are less likely to aggregate than wild-type counterparts. As used herein, these terms are not mutually exclusive or limiting, and it is entirely possible that a given variant has one or more modified characteristics of a wild-type protein.

[0049] In a particular aspect, the FGF21 protein variant is a biologically active variant of the wild-type FGF21 protein, which exhibits one or more biological activities of wild-type FGF21. Previously reported biological activities of wild-type FGF21 include, but are not limited to, the following: (i) FGF21 has been shown to induce insulin-independent glucose uptake; (ii) FGF21 has also been shown to improve hyperglycemia in a series of diabetic rodent models; (iii) transgenic mice overexpressing FGF21 were found to be resistant to diet-induced metabolic abnormalities and showed reduced body weight and fat mass, as well as enhanced insulin sensitivity (Badman, MK et al. (2007) Cell Metab [Cell Metabolism] 5, 426-37); (iv) administration of FGF21 to diabetic non-human primates results in a decrease in fasting glucose, triglycerides, insulin and glucagon levels, and a significant improvement in lipoprotein profiles, including an increase of nearly 80% in HDL cholesterol (Kharitonenkov, A. et al., 2007, Endocrinology [Endocrinology] 148, 774-81); (v) FGF21, as an important endocrine hormone, helps control the adaptation to the fasting state (Badman et al. 2009, Endocrinology [Endocrinology] 150, 4931; Inagaki et al., 2007, Cell Metabolism [Cell Metabolism] 5, 415); and (vi) FGF21 can regulate downstream markers, including but not limited to glucose or 2-deoxy-glucose uptake, pERK and other phosphorylated or acetylated proteins or NAD levels.

[0050] The term "native Fc" refers to a molecule or sequence comprising a non-antigen binding fragment sequence caused by digestion of a complete antibody or produced by other means, which is in a monomeric or multimeric form and may contain a hinge region. The original immunoglobulin source of natural Fc is preferably human and may be any immunoglobulin, although IgG1 and IgG2 are preferred. Natural Fc molecules consist of monomeric polypeptides that can be linked to dimer or multimeric form by covalent (i.e., disulfide bonds) and non-covalent association. The number of intermolecular disulfide bonds between monomer subunits of natural Fc molecules ranges from 1 to 4, depending on the class (e.g., IgG, IgA, and IgE) or subclass (e.g., IgG1, IgG2, IgG3, IgA1, and IgGA2). An example of natural Fc is a disulfide bonded dimer produced by papain digestion of IgG (see Elison et al., 1982, Nucleic Acids Res. [Nucleic Acids Research] 10: 4071-9). The term "native Fc" as used herein is generic to monomeric, dimeric and multimeric forms.

[0051] The term "Fc variant" refers to a molecule or sequence modified from a native Fc but still comprising a binding site for a salvage receptor FcRn (neonatal Fc receptor). International Publication Nos. WO 97 / 34631 and WO 96 / 32478 describe exemplary Fc variants, as well as interactions with salvage receptors, and are hereby incorporated by reference for this purpose. Therefore, the term "Fc variant" may include a humanized molecule or sequence from a non-human native Fc. In addition, the native Fc includes regions that can be removed because they provide structural features or biological activities that are not needed by the fusion molecule of the fusion protein of the present invention. Thus, the term "Fc variant" encompasses molecules or sequences that lack one or more native Fc sites or residues, or in which one or more Fc sites or residues are modified, which Fc sites or residues affect or participate in: (1) disulfide bond formation, (2) incompatibility with a selected host cell, (3) N-terminal heterogeneity after expression in a selected host cell, (4) glycosylation, (5) interaction with complement, (6) binding to Fc receptors other than salvage receptors, and / or (7) antibody-dependent cellular cytotoxicity (ADCC). Fc variants are described in further detail below.

[0052] The term "Fc domain" encompasses native Fc as defined above, as well as Fc variants and sequences. Like Fc variants and native Fc molecules, the term "Fc domain" includes molecules in monomeric or polymeric form that are digested from intact antibodies or otherwise produced. In some embodiments of the invention, the Fc domain can be fused to FGF21 or FGF21 mutants (including truncated forms of FGF21 or FGF21 mutants) via a covalent bond between, for example, the Fc domain and the FGF21 sequence. Such fusion proteins can form multimers via the association of the Fc domain, and these fusion proteins and their multimers are an aspect of the present invention.

[0053] As used herein, the term "modified Fc fragment" shall mean an Fc fragment of an antibody comprising a modified sequence. An Fc fragment is an antibody portion comprising CH2, CH3 and a portion of the hinge region. The modified Fc fragment can be derived from, for example, IgG1, IgG2, IgG3, or IgG4. FcLALA is a modified Fc fragment with a LALA mutation (L234A, L235A) that triggers ADCC with lower efficiency and weakly binds and activates human complement. Hessell et al. 2007 Nature [Nature] 449: 101-104. Other modifications to the Fc fragment are described in, for example, U.S. Patent No. 7,217,798, which is incorporated by reference for this purpose.

[0054] The term "acute myocardial infarction" refers to the death of the heart muscle due to interruption of the blood supply to a part of the heart (e.g., due to acute blockage of a coronary artery). The resulting ischemia and lack of oxygen may lead to damage or death (infarction) of the heart muscle tissue (myocardium) if left untreated for a long time.

[0055] The term "atherosclerosis" is a vascular disease characterized by irregularly distributed lipid deposits in the intima of large and medium-sized arteries, resulting in narrowing of the arterial lumen and eventual progression to fibrosis and calcification. The lesions are usually focal and progress slowly and intermittently. Restriction of blood flow is responsible for most clinical manifestations, which vary with the distribution and severity of the lesions.

[0056] The term "cardiovascular disease" refers to diseases related to the heart or blood vessels.

[0057] The term "cardiovascular risk" refers to a combination of factors associated with a higher probability of a cardiovascular event (e.g., stroke or heart attack). Such factors may include, but are not limited to, weight, BMI, cholesterol levels, blood pressure, triglyceride levels, diet, exercise habits, age, sex, family history, obesity, diabetes, and / or other metabolic factors. Guidelines for assessing and managing cardiovascular risk have been described. For example, the American College of Cardiology (ACC) and the American Heart Association (AHA) collaborate with the National Heart, Lung, and Blood Institute (NHLBI) and stakeholders and professional organizations to develop guidelines for assessing cardiovascular risk, changing lifestyles to reduce cardiovascular risk, managing adult blood cholesterol, and managing clinical practice guidelines for overweight and obesity in adults (Stone et al., 2014, Journal of the American College of Cardiology [Journal of the American College of Cardiology], 63 (25, Part B) 2889-2934; DOI: 10.1016 / j.jacc.2013.11.002, which is hereby incorporated by reference for this purpose). The World Health Organization also provides guidelines for assessing cardiovascular risk, for example, individuals with persistently elevated blood pressure ≥160 / 100 mm Hg, blood cholesterol ≥8 mmol / 1, established ischemic heart disease, or diabetes with concomitant renal disease.

[0058] The term "coronary heart disease" (also called coronary artery disease) is the narrowing of the small blood vessels that supply blood and oxygen to the heart.

[0059] The terms "diabetes" and "diabetic" refer to a progressive disease of carbohydrate metabolism involving insufficient production or utilization of insulin, typically characterized by hyperglycemia and glycosuria. The terms "prediabetes" and "prediabetic" refer to a state in which the subject does not have the characteristics, symptoms, etc. typically observed in diabetes but has characteristics, symptoms, etc. that may progress to diabetes if not treated. The presence of these conditions can be determined using, for example, a fasting plasma glucose (FPG) test, an oral glucose tolerance test (OGTT), or a hemoglobin A1c (HbA1c) test. For both the FPG test and the OGTT, both typically require the subject to fast for at least 8 hours before starting the test. In the FPG test, the subject's blood sugar is measured after the end of the fast; typically, the subject fasts overnight and blood sugar is measured in the morning before the subject eats. Healthy subjects typically have FPG concentrations between about 90 and about 100 mg / dl, subjects with "prediabetes" typically have FPG concentrations between about 100 to about 125 mg / dl, and subjects with "diabetes" typically have FPG levels above about 126 mg / dl. In an OGTT, the blood sugar of the subject is measured after fasting, and again two hours after drinking a glucose-rich beverage. Two hours after consuming a glucose-rich beverage, healthy subjects typically have blood sugar concentrations below about 140 mg / dl, prediabetic subjects typically have blood sugar concentrations of about 140 to about 199 mg / dl, and diabetic subjects typically have blood sugar concentrations of about 200 mg / dl or more. Although the above blood sugar values ​​are related to human subjects, the proportions of normal blood sugar, moderate hyperglycemia, and significant hyperglycemia in mouse subjects are different. After four hours of fasting, healthy mouse subjects typically have FPG concentrations between about 100 mg / dl and about 150 mg / dl, mouse subjects with "prediabetes" typically have FPG concentrations between about 175 mg / dl and about 250 mg / dl, and mouse subjects with "diabetes" typically have FPG concentrations above about 250 mg / dl. For HbA1c testing, HbA1c levels <5.7% are generally considered to be within the normal range, HbA1c levels in the range of 5.7%-6.4% are generally considered to be prediabetic levels, and HbA1c levels of 6.5% or above are generally considered to be diabetic levels (see, e.g., American Diabetes Association (ADA), Practice Guideline, Diabetes Care, 2018 Supplement 1, which is hereby incorporated by reference for this purpose).

[0060] The term "dyslipidemia" is a disorder of lipoprotein metabolism involving the overproduction or deficiency of lipoproteins. Dyslipidemia may manifest as elevated total cholesterol, elevated low-density lipoprotein (LDL) cholesterol and triglyceride concentrations, and decreased high-density lipoprotein (HDL) cholesterol concentrations in the blood.

[0061] The term "glucose intolerance" or "impaired fasting glucose" (IFG) or "impaired glucose tolerance" (IGT) is a prediabetic state of impaired blood glucose metabolism associated with an increased risk of cardiovascular pathology. The prediabetic condition prevents the subject from efficiently transferring glucose into cells and using it as an efficient fuel source, resulting in elevated glucose levels in the blood and a degree of insulin resistance.

[0062] The term "heart failure" (also called congestive heart failure) is a condition in which the heart can no longer pump enough blood to the rest of the body.

[0063] The term "hyperglycemia" (also known as high blood sugar) refers to a condition characterized by higher than normal amounts of glucose (a sugar) in the blood. Hyperglycemia can be diagnosed using methods known in the art, including measuring fasting blood sugar levels.

[0064] The term "hypertension" or "high blood pressure" refers to a condition in which systemic arterial blood pressure is temporarily or persistently elevated to a level that may induce cardiovascular damage or other adverse consequences. Hypertension is mandatorily defined as a systolic blood pressure (SBP) above 140 mmHg or a diastolic blood pressure (DBP) above 90 mmHg, but may also be defined according to published clinical guidelines. For example, the following blood pressure guidelines have been described: normal SBP < 120 mmHg or DBP < 80 mmHg; elevated SBP or DBP < 80 mmHg in the range of 120-129 mmHg; hypertension (HTN) stage 1: SBP in the range of 130-139 mmHg or DBP 80-89 mmHg; and HTN: SBP ≥ 140 or DBP ≥ 90.

[0065] The term "hypertriglyceridemia" refers to a high blood level of triglycerides (congestion). Elevated triglyceride levels are associated with atherosclerosis and are susceptible to cardiovascular disease (e.g., contributing to increased cardiovascular disease risk) even in the absence of hypercholesterolemia (high cholesterol levels). Typically, serum triglyceride levels in the range of 150 to 199 mg / dL [1.70 mmol / L to 2.25 mmol / L] are considered to be critically high serum triglyceride levels; serum triglyceride levels in the range of 200 to 499 mg / dL [2.26 mmol / L to 5.64 mmol / L] are considered to be high serum triglyceride levels; and serum triglyceride levels in the range of 500 mg / dL [5.65 mmol / L] or higher are considered to be very high triglyceride levels. In some aspects, these general guidelines can be adjusted based on updated clinical guidelines for clinicians.

[0066] The term "hypoglycemia" (also called low blood sugar) occurs when blood sugar levels drop too low to provide enough energy for physical activity.

[0067] The term "hyperinsulinemia" is defined as higher than normal levels of insulin in the blood.

[0068] The term "insulin resistance" is defined as a state in which normal amounts of insulin produce a subnormal biologic response.

[0069] The term "kidney disease" or nephropathy is any disease of the kidneys. Diabetic nephropathy refers to kidney damage caused by diabetes; in severe cases, it can lead to kidney failure, a leading cause of morbidity and mortality in people with type 1 or type 2 diabetes.

[0070] The term "metabolic syndrome" refers to a cluster of conditions (increased blood pressure, high blood sugar, excess body fat around the waist, and abnormal cholesterol or triglyceride levels) that occur together and increase the risk of heart disease, stroke, and diabetes. For example, in most men, excess body fat around the waist is associated with a waist circumference of 40 inches or more; high blood sugar is associated with a fasting blood sugar / glucose level of at least 110 milligrams per deciliter (mg / dl); high triglycerides are associated with levels of at least 150 mg / dL in the bloodstream; low HDL is associated with levels less than 40 mg / dl; and elevated blood pressure is associated with levels of 130 / 85 mmHg or higher.

[0071] The term "myocardial infarction" (MI), also known as heart attack, is defined as the irreversible death (necrosis) of the heart muscle due to a prolonged lack of oxygen supply (ischemia).

[0072] The term "NAFLD" or "non-alcoholic fatty liver disease" is defined as a condition in which excess fat is stored in the liver. This fat accumulation is not caused by heavy alcohol use. When heavy alcohol use causes fat accumulation in the liver, the condition is called alcoholic liver disease. Typically, there are two types of NAFLD: simple fatty liver and non-alcoholic steatohepatitis (NASH).

[0073] The term "NASH" or "non-alcoholic steatohepatitis" is defined as a form of non-alcoholic fatty liver disease (NAFLD) in which, in addition to liver fat, patients also have hepatitis - inflammation of the liver - liver cell damage. Inflammation and liver cell damage can lead to fibrosis or scarring of the liver. In some cases, NASH may lead to cirrhosis or liver cancer.

[0074] The term "simple fatty liver" (also called nonalcoholic fatty liver (NAFL)) is a form of NAFLD in which patients have liver fat, but little or no inflammation or liver cell damage. Simple fatty liver typically does not progress to cause liver damage or complications.

[0075] The term "pancreatitis" is inflammation of the pancreas.

[0076] "Obesity" is defined as abnormal or excessive fat accumulation that poses a risk to health. A crude population measure of obesity is body mass index (BMI = weight (kg) / [height (m)] 2 ), which is a person's weight in kilograms divided by the square of his or her height in meters. People with a BMI of 30 or more are generally considered obese. In general, for example, people with a BMI equal to or greater than 25 for Caucasians are considered overweight, but the guidelines can be adjusted for racial differences. For example, by adjusting for race, for Asian individuals, a BMI ≥ 27.5 can be considered obese (WHO Expert Consultation, 2004, Lancet, 363(9403): 157-63). Asian individuals can be, for example, subjects of Asian descent or Asian ancestry.

[0077] The term "peripheral arterial disease" or "PAD" refers to a condition characterized by narrowing of the peripheral arteries (most commonly arteries from the legs) to the legs, stomach, arms, and head. PAD is similar to coronary artery disease (CAD) in that both PAD and CAD are caused by atherosclerosis, which narrows and blocks arteries in various critical areas of the body. The narrowing of the arteries is typically due to plaque, which can harden and narrow the arteries over time, restricting the flow of oxygen-rich blood to organs and other parts of the body.

[0078] The term "stroke" is any acute clinical event associated with impaired cerebral circulation lasting more than 24 hours, while shorter durations are considered transient ischemic attacks (TIAs). Stroke involves irreversible brain damage, and the type and severity of symptoms depend on the location and extent of brain tissue to which circulation is impaired.

[0079] The term "type 1 diabetes" or "T1DM" is a condition characterized by high blood sugar levels caused by a lack of insulin. The condition occurs when the body's immune system attacks the insulin-producing beta cells in the pancreas and destroys them. The pancreas then produces little or no insulin.

[0080] The term "type 2 diabetes" or "type 2 diabetes" or "T2D" or "T2DM" refers to a metabolic disease that causes sugar to accumulate in the bloodstream and is characterized by high blood sugar levels caused by a lack of insulin or the body's inability to use insulin effectively. Blood sugar levels are measured in milligrams per deciliter (mg / dL) or mmol / L.

[0081] The term "management" or "manage" is understood as the management and care of a patient for the purpose of combating a disease, condition or disorder, which does not result in a cure but does result in relief of one or more symptoms of the disease, condition or disorder and / or a reduction in hospital stay.

[0082] The term "prevention" or "preventing" refers to the prophylactic administration to healthy subjects or subjects at risk of developing a disease, condition or disorder (e.g., a subject considered to be pre-diabetic) to prevent the development of one or more conditions mentioned herein. In addition, the term "prevention" may also include prophylactic administration to patients who are at a pre-stage of the condition to be treated.

[0083] The term "treatment" or "treat" or "treating" is understood as the management and care of a patient for the purpose of combating a disease, condition or disorder, for example, reducing or ameliorating the progression, severity and / or duration of the disease, condition or disorder. In certain aspects, in the context of a disease, condition, or disorder that can be treated by administering the FGF21 or FGF21 variant proteins provided herein, the treatment used herein can include, but is not limited to, one or more of the following: (i) lowering triglyceride levels (blood / serum triglyceride levels), e.g., to a manageable range or normal range as determined by a clinician or clinical guidelines; (ii) reducing body weight, e.g., to a manageable range or normal range as determined by a clinician or clinical guidelines, or by at least about 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60% or more; (iii) increasing insulin-independent glucose uptake; (iv) lowering blood / serum glucose levels (e.g., to a manageable range or normal range as determined by a clinician or clinical guidelines, or by at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60% or more; (e.g., by at least about 5%, 10%, 20%, 30%, 40%, 45%, 50%, 55%, 60%, 70%, 80%, 90%, or 95% or more); (v) improve lipoprotein profile; (vi) increase HDL cholesterol (HDL-C) levels (e.g., by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more); (vii) reduce LDL cholesterol (LDL-C) and / or total cholesterol (viii) reducing liver fat content, for example, to within a manageable range or normal range as determined by a clinician or clinical guidelines, or by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more; and (ix) reducing the incidence of complications.

[0084] The term "therapeutically effective amount" or "effective amount" refers to the amount of a drug or therapeutic agent that will elicit the desired biological and / or medical response of a tissue, system or animal (including humans) being sought by a researcher or clinician. In the context of the FGF21 and FGF21 variant proteins provided herein, the desired biological and / or medical response can be one or more of the following: (i) lowering triglyceride levels (blood / serum triglyceride levels), for example, by at least about 5%, 10%, 20%, 30%, 40%, 45%, 50%, 55%, 60%, 70%, 80%, 90%, or 95% or more; (ii) reducing body weight, for example, by at least about 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60% or more; (iii) increasing insulin-independent glucose uptake; (iv) lowering blood / serum glucose (v) improve lipoprotein profile; (vi) increase HDL cholesterol (HDL-C) levels (e.g., increase by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 95% or more); (vii) reduce LDL cholesterol (LDL-C) and / or total cholesterol (total C), for example, by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more; and (viii) reduce liver fat content, for example, by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more. In certain embodiments, triglyceride levels can be reduced by at least about 40% or at least about 50%. In certain embodiments, triglyceride levels may be reduced by at least about 40%. In certain embodiments, triglyceride levels may be reduced by at least about 50%. In some embodiments, liver fat content may be reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more. In some embodiments, liver fat content may be reduced by at least about 5%. In some embodiments, liver fat content may be reduced by at least about 10%. In some embodiments, liver fat content may be reduced by at least about 20%. In some embodiments, liver fat content may be reduced by at least about 30%. In some embodiments, liver fat content may be reduced by at least about 40%. In some embodiments, liver fat content may be reduced by at least about 50%. In some embodiments, liver fat content may be reduced to a manageable range or normal range as determined by a clinician or clinical guidelines.

[0085] As used herein, the singular forms "a", "an", and "the" include plural referents unless the content clearly dictates otherwise. Thus, for example, reference to "an antibody" includes a mixture of two or more such antibodies.

[0086] As used herein, the term "about" or "approximately" refers to + / - 20%, more preferably + / - 10%, or still more preferably + / - 5% of a value.

[0087] The terms "polypeptide" and "protein" are used interchangeably and refer to a polymeric form of amino acids of any length, which may include coded and non-coded amino acids, naturally occurring and non-naturally occurring amino acids, chemically or biochemically modified or derivatized amino acids, and polypeptides with modified peptide backbones. The term includes fusion proteins, including but not limited to, fusion proteins with heterologous amino acid sequences, fusions with heterologous and homologous leader sequences (with or without an N-terminal methionine residue); immunolabeled proteins; etc.

[0088] As used herein, the term "fragment" refers to a physically continuous portion of the primary structure of a biomolecule. In the case of a protein, a portion is defined by a continuous portion of the amino acid sequence of the protein and refers to at least 3-5 amino acids, at least 8-10 amino acids, at least 11-15 amino acids, at least 17-24 amino acids, at least 25-30 amino acids, and at least 30-45 amino acids. In the case of an oligonucleotide, a portion is defined by a continuous portion of the nucleic acid sequence of the oligonucleotide and refers to at least 9-15 nucleotides, at least 18-30 nucleotides, at least 33-45 nucleotides, at least 48-72 nucleotides, at least 75-90 nucleotides, and at least 90-130 nucleotides. In some embodiments, a portion of a biomolecule has biological activity. In the context of an FGF21 polypeptide, an FGF21 polypeptide fragment does not include the complete FGF21 polypeptide sequence shown in SEQ ID NO: 3.

[0089] The terms "individual", "subject", "host" and "patient" are used interchangeably and refer to any subject, particularly a human, in need of diagnosis, treatment or therapy. Other subjects may include cows, dogs, cats, guinea pigs, rabbits, rats, mice, horses, etc. In some preferred embodiments, the subject is a human.

[0090] As used herein, the term "sample" refers to biological material from a patient. The samples assayed by the present invention are not limited to any particular type. As non-limiting examples, samples include supernatants or extracts of single cells, multiple cells, tissues, tumors, biological fluids, biomolecules, or any of the foregoing. Examples include tissue removed for biopsy, tissue removed during resection, blood, urine, lymphatic tissue, lymph fluid, cerebrospinal fluid, mucus, and fecal samples. The sample used will vary depending on the assay format, detection method, and properties of the tumor, tissue, cell, or extract to be assayed. Methods for preparing samples are well known in the art and can be easily adjusted to obtain samples compatible with the methods used.

[0091] The term "pharmaceutically acceptable carrier" refers to a carrier for administering therapeutic agents such as antibodies or polypeptides, genes and other therapeutic agents. The term refers to any drug carrier that does not itself induce the production of antibodies that are harmful to the individual receiving the composition, and it can be administered without excessive toxicity. Suitable carriers can be large, slowly metabolized macromolecules, such as proteins, polysaccharides, polylactic acid, polyglycolic acid, polymeric amino acids, amino acid copolymers, lipid aggregates and / or inactive viral particles. Such carriers are well known to those of ordinary skill in the art. The pharmaceutically acceptable carrier in the therapeutic composition can include liquids, such as water, saline, glycerol and / or ethanol. Auxiliary substances such as wetting agents or emulsifiers, pH buffer substances, etc. can also appear in such carriers.

[0092] Treatment

[0093] Provided herein are methods and pharmaceutical compositions for treating, preventing or managing metabolic disorders or cardiovascular disorders (e.g., alleviating one or more symptoms thereof), comprising administering human FGF21 protein variants (e.g., Fc-FGF21 mutant fusion proteins, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose ranging from 100 mg to 600 mg, for example, once a day, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month). In a specific aspect, the human FGF21 protein variant (e.g., Fc-FGF21 mutant fusion protein, such as V103 (SEQ ID NO: 11)) is administered at a dose ranging from 200 mg to 400 mg. In a specific aspect, the human FGF21 protein variant (e.g., Fc-FGF21 mutant fusion protein, such as V103 (SEQ ID NO: 11)) is administered at a dose of about 200 mg, 250 mg, 300 mg, 350 mg, or 400 mg, for example, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month). In a specific aspect, the human FGF21 protein variant (e.g., Fc-FGF21 mutant fusion protein, such as V103 (SEQ ID NO: 11)) is administered once every 4 weeks (or once a month). In certain aspects, human FGF21 protein variants (e.g., V103 (SEQ ID NO: 11)) are administered once every 3 weeks, once every 2 weeks, or once a week.

[0094] In a specific aspect, provided herein is a method for treating, preventing or managing a metabolic disorder or a cardiovascular disorder (e.g., obesity or hypertriglyceridemia) (e.g., alleviating one or more symptoms thereof), the method comprising administering a human FGF21 protein variant (e.g., Fc-FGF21 mutant fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose ranging from 100 mg to 600 mg once a week or once every two weeks. In a specific aspect, provided herein is a method for treating, preventing or managing a metabolic disorder or a cardiovascular disorder (e.g., obesity or hypertriglyceridemia) (e.g., alleviating one or more symptoms thereof), the method comprising administering a human FGF21 protein variant (e.g., Fc-FGF21 mutant fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose ranging from 100 mg to 600 mg once every three weeks or once every four weeks (or once a month).

[0095] In a specific aspect, provided herein is a method for treating, preventing or managing a metabolic disorder or a cardiovascular disorder (e.g., obesity or hypertriglyceridemia) (e.g., alleviating one or more symptoms thereof), the method comprising administering a human FGF21 protein variant (e.g., Fc-FGF21 mutant fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose ranging from 200 mg to 400 mg once a week or once every two weeks. In a specific aspect, provided herein is a method for treating, preventing or managing a metabolic disorder or a cardiovascular disorder (e.g., obesity or hypertriglyceridemia) (e.g., alleviating one or more symptoms thereof), the method comprising administering a human FGF21 protein variant (e.g., Fc-FGF21 mutant fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose ranging from 200 mg to 400 mg once every three weeks or once every four weeks (or once a month).

[0096] In a specific aspect, provided herein is a method for treating, preventing or managing a metabolic disorder or a cardiovascular disorder (e.g., obesity or hypertriglyceridemia) (e.g., alleviating one or more symptoms thereof), the method comprising administering a human FGF21 protein variant (e.g., Fc-FGF21 mutant fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose in the range of 100 mg to 300 mg once a week or once every two weeks. In a specific aspect, provided herein is a method for treating, preventing or managing a metabolic disorder or a cardiovascular disorder (e.g., obesity or hypertriglyceridemia) (e.g., alleviating one or more symptoms thereof), the method comprising administering a human FGF21 protein variant (e.g., Fc-FGF21 mutant fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose in the range of 100 mg to 300 mg once every three weeks or once every four weeks (or once a month).

[0097] In a specific aspect, provided herein is a method for treating, preventing or managing a metabolic disorder or a cardiovascular disorder (e.g., obesity or hypertriglyceridemia) (e.g., alleviating one or more symptoms thereof), the method comprising administering a human FGF21 protein variant (e.g., an Fc-FGF21 mutant fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of at least about 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, or 350 mg once a week or once every two weeks. In one specific aspect, provided herein is a method for treating, preventing or managing a metabolic disorder or a cardiovascular disorder (e.g., obesity or hypertriglyceridemia) (e.g., alleviating one or more symptoms thereof), the method comprising administering a human FGF21 protein variant (e.g., an Fc-FGF21 mutant fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of at least about 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, or 350 mg once every three weeks or once every four weeks (or once a month).

[0098] In one aspect, non-limiting examples of metabolic or cardiovascular disorders to be treated, prevented or managed by the methods provided herein include obesity, hypertriglyceridemia, dyslipidemia, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), diabetes (e.g., type 2 diabetes (T2DM) and type 1 diabetes (T1DM)), diseases associated with severe inactivating mutations in the insulin receptor, pancreatitis, insulin resistance, hyperinsulinemia, glucose intolerance, hyperglycemia, metabolic syndrome, hypertension, cardiovascular disease, acute myocardial infarction, atherosclerosis, peripheral arterial disease, stroke, heart failure, coronary heart disease, kidney disease, diabetic complications, neuropathy and gastroparesis.

[0099] In one aspect, non-limiting examples of metabolic or cardiovascular disorders to be treated, prevented, or managed by the methods provided herein include obesity, hypertriglyceridemia and risk of heart disease, insulin resistance (e.g., patients with genetic mutations of the insulin receptor) and lipodystrophy, diabetes (e.g., T1DM or T2DM), and non-alcoholic fatty liver disease (NAFLD) / non-alcoholic steatohepatitis (NASH).

[0100] In one aspect, provided herein is a method for treating, preventing or managing a metabolic disorder (e.g., obesity) or a cardiovascular disorder, the method comprising administering (e.g., subcutaneously (sc)) a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose in the range of 100 mg to 600 mg (or more specifically, in the range of 100 mg to 300 mg or 200 mg to 400 mg), for example once a day, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month). In one aspect, provided herein is a method for treating, preventing or managing a metabolic disorder (e.g., obesity) or a cardiovascular disorder, the method comprising administering (e.g., sc) a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of about 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg, for example, once a day, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month). In one specific aspect, provided herein is a method for treating, preventing or managing a metabolic disorder (e.g., obesity) or a cardiovascular disorder, the method comprising administering subcutaneously (e.g., sc) a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of about 300 mg every 4 weeks (monthly). In one specific aspect, provided herein is a method for treating, preventing or managing a metabolic disorder (e.g., obesity) or a cardiovascular disorder, the method comprising administering subcutaneously (e.g., sc) a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of about 300 mg every 3 weeks. In one specific aspect, provided herein is a method for treating, preventing or managing a metabolic disorder (e.g., obesity) or a cardiovascular disorder, the method comprising administering subcutaneously (e.g., sc) a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of about 300 mg every 2 weeks.In a specific aspect, provided herein is a method for treating, preventing or managing a metabolic disorder (e.g., obesity) or a cardiovascular disorder, the method comprising administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) subcutaneously to a subject in need thereof at a dose in the range of about 200 mg to 250 mg every 3 weeks. In a specific aspect, the method provided herein is used to treat a human subject with hypercholesterolemia (e.g., primary hypercholesterolemia) or dyslipidemia (e.g., mixed dyslipidemia). In a specific aspect, the method provided herein is used to treat a human subject with hypertriglyceridemia (particularly severe hypertriglyceridemia). In certain aspects, the method provided herein is used to treat a human subject with type 2 diabetes or an obese human subject. In a specific aspect, the method provided herein is used to treat a human subject aged 18 to 55 years, who, prior to administration of the human FGF21 protein variant, has (i) 30-45 kg / m. 2 BMI within the range of (including the end value) (wherein for Asian subjects, race-adjusted greater than or equal to 27.5 kg / m 2 BMI), and (ii) triglyceride levels in the range of 150 to 500 mg / dL (1.69-5.65 mmol / L).

[0101] In a specific aspect, provided herein is a method for treating, preventing or managing hypercholesterolemia, dyslipidemia (e.g., mixed dyslipidemia) or hypertriglyceridemia (e.g., alleviating one or more symptoms thereof), the method comprising administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose in the range of 100 mg to 600 mg (or more specifically, at a dose in the range of 100 mg to 300 mg or 200 mg to 400 mg), for example once a day, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month). In a specific aspect, provided herein is a method for treating, preventing or managing hypercholesterolemia, dyslipidemia (e.g., mixed dyslipidemia) or hypertriglyceridemia (e.g., alleviating one or more symptoms thereof), the method comprising administering about 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg of 1% glycerol. The human FGF21 protein variant (e.g., Fc fusion protein, such as V103 (SEQ ID NO: 11)) is administered to a subject in need thereof at a dosage of, for example, once a day, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month). In a certain aspect, provided herein is a method for treating, preventing or managing hypercholesterolemia, dyslipidemia (e.g., mixed dyslipidemia) or hypertriglyceridemia (e.g., alleviating one or more symptoms thereof), the method comprising administering a human FGF21 protein variant (e.g., Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof subcutaneously at a dosage of about 300 mg once every 4 weeks (or once a month). In a certain aspect, provided herein is a method for treating, preventing or managing hypercholesterolemia, dyslipidemia (e.g., mixed dyslipidemia) or hypertriglyceridemia (e.g., alleviating one or more symptoms thereof), the method comprising subcutaneously administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of about 300 mg once every 3 weeks.In a certain aspect, provided herein is a method for treating, preventing or managing hypercholesterolemia, dyslipidemia (e.g., mixed dyslipidemia) or hypertriglyceridemia (e.g., alleviating one or more symptoms thereof), the method comprising subcutaneously administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of about 300 mg once every 2 weeks. In a certain aspect, provided herein is a method for treating, preventing or managing hypercholesterolemia, dyslipidemia (e.g., mixed dyslipidemia) or hypertriglyceridemia (e.g., alleviating one or more symptoms thereof), the method comprising subcutaneously administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of about 300 mg once a week. In a certain aspect, provided herein is a method for treating, preventing or managing hypercholesterolemia, dyslipidemia (e.g., mixed dyslipidemia) or hypertriglyceridemia (e.g., alleviating one or more symptoms thereof), the method comprising administering subcutaneously to a subject in need thereof a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) at a dose in the range of about 200 mg to 250 mg once every 3 weeks or once every 2 weeks. In a specific aspect, the method provided herein is used to treat a human subject with hypercholesterolemia (e.g., primary hypercholesterolemia) or dyslipidemia (e.g., mixed dyslipidemia). In a specific aspect, the method provided herein is used to treat a human subject with hypertriglyceridemia (particularly severe hypertriglyceridemia). In certain aspects, the method provided herein is used to treat a human subject with type 2 diabetes or an obese human subject. In a specific aspect, the method provided herein is used to treat a human subject aged 18 to 55 years, who, prior to administration of the human FGF21 protein variant, has (i) 30-45 kg / m. 2 BMI within the range of (including the end value) (wherein for Asian subjects, race-adjusted greater than or equal to 27.5 kg / m 2 BMI), and (ii) triglyceride levels in the range of 150 to 500 mg / dL (1.69-5.65 mmol / L).

[0102] In a specific aspect, provided herein is a method for reducing body weight (e.g., reducing body weight by at least 5%, 6%, 7%, 8%, 9%, 10%, 15%, or 20% or more), the method comprising administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose in the range of 100 mg to 600 mg (or more specifically, at a dose in the range of 200 mg to 400 mg or 100 mg to 300 mg), for example, once a day, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month). In a certain aspect, provided herein is a method for reducing body weight (e.g., reducing body weight by at least 5%, 6%, 7%, 8%, 9%, 10%, 15%, or 20% or more), the method comprising administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose in the range of 100 mg to 600 mg (or more specifically, at a dose in the range of 200 mg to 400 mg or 100 mg to 300 mg), for example, once a day, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month). 140mg, 150mg, 160mg, 170mg, 180mg, 190mg, 200mg, 210mg, 220mg, 230mg, 240mg, 250mg, 260mg, 270mg, 280mg, 290mg, 300mg, 310mg, 320mg, 330mg, 340mg, or 350mg, for example, once a day, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month) to a subject in need thereof. In a certain aspect, provided herein is a method for reducing body weight (e.g., reducing body weight by at least 5%, 6%, 7%, 8%, 9%, 10%, 15%, or 20% or more), the method comprising administering human FGF21 protein variant (e.g., Fc fusion protein, such as V103 (SEQ ID NO: 11)) subcutaneously to a subject in need thereof at a dose of about 300 mg once every 4 weeks (or once a month). In a certain aspect, provided herein is a method for reducing body weight (e.g., reducing body weight by at least 5%, 6%, 7%, 8%, 9%, 10%, 15%, or 20% or more), the method comprising administering human FGF21 protein variant (e.g., Fc fusion protein, such as V103 (SEQ ID NO: 11)) subcutaneously to a subject in need thereof at a dose of about 300 mg once every 3 weeks. In one aspect, provided herein is a method for reducing body weight (e.g., reducing body weight by at least 5%, 6%, 7%, 8%, 9%, 10%, 15%, or 20% or more), the method comprising subcutaneously administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of about 300 mg once every 2 weeks.In a certain aspect, provided herein is a method for reducing body weight (e.g., reducing body weight by at least 5%, 6%, 7%, 8%, 9%, 10%, 15%, or 20% or more), the method comprising administering human FGF21 protein variant (e.g., Fc fusion protein, such as V103 (SEQ ID NO: 11)) subcutaneously to a subject in need thereof at a dose of about 300 mg once a week. In a certain aspect, provided herein is a method for reducing body weight (e.g., reducing body weight by at least 5%, 6%, 7%, 8%, 9%, 10%, 15%, or 20% or more), the method comprising administering human FGF21 protein variant (e.g., Fc fusion protein, such as V103 (SEQ ID NO: 11)) subcutaneously to a subject in need thereof at a dose in the range of about 200 mg to 250 mg once every 3 weeks or once every 2 weeks. In a specific aspect, the methods provided herein are used to treat human subjects with hypercholesterolemia (e.g., primary hypercholesterolemia) or dyslipidemia (e.g., mixed dyslipidemia). In certain aspects, provided herein is a method for treating a human subject suffering from hypertriglyceridemia (particularly severe hypertriglyceridemia). In certain aspects, provided herein is a method for treating a human subject suffering from type 2 diabetes or an obese human subject. In certain aspects, provided herein is a method for treating a human subject aged 18 to 55 years, and when screening before applying human FGF21 protein variants, the human subject has (i) at 30-45kg / m. 2 BMI within the range of (including the end value) (wherein for Asian subjects, race-adjusted greater than or equal to 27.5 kg / m 2 BMI), and (ii) triglyceride levels in the range of 150 to 500 mg / dL (1.69-5.65 mmol / L).

[0103] In a specific aspect, provided herein is a method for treating, preventing or managing obesity (e.g., alleviating one or more symptoms thereof), the method comprising administering to a subject in need thereof a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) at a dose in the range of 100 mg to 600 mg (or more specifically, at a dose in the range of 200 mg to 400 mg or 100 mg to 300 mg), for example once a day, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month). In a specific aspect, provided herein is a method for treating, preventing or managing obesity (e.g., alleviating one or more symptoms thereof), the method comprising administering to a subject in need thereof a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) at a dose of about 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg, for example, once a day, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month). In a certain aspect, provided herein is a method for treating, preventing or managing obesity (e.g., alleviating one or more symptoms thereof), the method comprising subcutaneously administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of about 300 mg once every 4 weeks (once a month). In a certain aspect, provided herein is a method for treating, preventing or managing obesity (e.g., alleviating one or more symptoms thereof), the method comprising subcutaneously administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of about 300 mg once every 3 weeks. In a certain aspect, provided herein is a method for treating, preventing or managing obesity (e.g., alleviating one or more symptoms thereof), the method comprising subcutaneously administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of about 300 mg once every 2 weeks.In a certain aspect, provided herein is a method for treating, preventing or managing obesity (e.g., alleviating one or more symptoms thereof), the method comprising administering human FGF21 protein variant (e.g., Fc fusion protein, such as V103 (SEQ ID NO: 11)) subcutaneously to a subject in need thereof at a dose of about 300 mg once a week. In a certain aspect, provided herein is a method for treating, preventing or managing obesity (e.g., alleviating one or more symptoms thereof), the method comprising administering human FGF21 protein variant (e.g., Fc fusion protein, such as V103 (SEQ ID NO: 11)) subcutaneously to a subject in need thereof at a dose in the range of about 200 mg to 250 mg once every 3 weeks or once every 2 weeks. In a specific aspect, the method provided herein is used to treat a human subject with hypercholesterolemia (e.g., primary hypercholesterolemia) or dyslipidemia (e.g., mixed dyslipidemia). In a specific aspect, the method provided herein is used to treat a human subject with hypertriglyceridemia (particularly severe hypertriglyceridemia). In certain aspects, the method provided herein is used to treat a human subject suffering from type 2 diabetes or an obese human subject. In a specific aspect, the method provided herein is used to treat a human subject aged 18 to 55 years old, and when screening before using human FGF21 protein variants, the human subject has (i) at 30-45kg / m. 2 BMI within the range of (including the end value) (wherein for Asian subjects, race-adjusted greater than or equal to 27.5 kg / m 2 BMI), and (ii) triglyceride levels in the range of 150 to 500 mg / dL (1.69-5.65 mmol / L).

[0104] In a specific aspect, provided herein is a method for treating, preventing or managing NASH (e.g., alleviating one or more symptoms thereof), the method comprising administering to a subject in need thereof a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) at a dose in the range of 100 mg to 600 mg (or more specifically, at a dose in the range of 200 mg to 400 mg or 100 mg to 300 mg), for example once a day, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month). In a specific aspect, provided herein is a method for treating, preventing or managing NASH (e.g., alleviating one or more symptoms thereof), the method comprising administering to a subject in need thereof a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) at a dose of about 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg, for example, once a day, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month). In a certain aspect, provided herein is a method for treating, preventing or managing NASH (e.g., alleviating one or more symptoms thereof), the method comprising subcutaneously administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of about 300 mg once every 4 weeks (or once a month). In a certain aspect, provided herein is a method for treating, preventing or managing NASH (e.g., alleviating one or more symptoms thereof), the method comprising subcutaneously administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of about 300 mg once every 3 weeks. In a certain aspect, provided herein is a method for treating, preventing or managing NASH (e.g., alleviating one or more symptoms thereof), the method comprising subcutaneously administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of about 300 mg once every 2 weeks.In a certain aspect, provided herein is a method for treating, preventing or managing NASH (e.g., alleviating one or more symptoms thereof), the method comprising subcutaneously administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of about 300 mg once a week. In a certain aspect, provided herein is a method for treating, preventing or managing NASH (e.g., alleviating one or more symptoms thereof), the method comprising subcutaneously administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose in the range of about 200 mg to 250 mg once every 3 weeks or once every 2 weeks. In a specific aspect, the method provided herein is used to treat a human subject with hypercholesterolemia (e.g., primary hypercholesterolemia) or dyslipidemia (e.g., mixed dyslipidemia). In a specific aspect, the method provided herein is used to treat a human subject with hypertriglyceridemia (particularly severe hypertriglyceridemia). In certain aspects, the methods provided herein are used to treat a human subject suffering from type 2 diabetes or an obese human subject. In specific aspects, the methods provided herein are used to treat a human subject aged 18 to 55 years, and before administering human FGF21 protein variants, the human subject has (i) 30-45 kg / m. 2 BMI within the range of (including the end value) (wherein for Asian subjects, race-adjusted greater than or equal to 27.5 kg / m 2 BMI), and (ii) triglyceride levels in the range of 150 to 500 mg / dL (1.69-5.65 mmol / L).

[0105] In a specific aspect, provided herein is a method for treating, preventing or managing NAFLD (e.g., alleviating one or more symptoms thereof), the method comprising administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose in the range of 100 mg to 600 mg (or more specifically, at a dose in the range of 200 mg to 400 mg or 100 mg to 300 mg), for example once a day, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month). In a specific aspect, provided herein is a method for treating, preventing or managing NAFLD (e.g., alleviating one or more symptoms thereof), the method comprising administering to a subject in need thereof a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) at a dose of about 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg, for example, once a day, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month). In a certain aspect, provided herein is a method for treating, preventing or managing NAFLD (e.g., alleviating one or more symptoms thereof), the method comprising subcutaneously administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of about 300 mg once every 4 weeks (or once a month). In a certain aspect, provided herein is a method for treating, preventing or managing NAFLD (e.g., alleviating one or more symptoms thereof), the method comprising subcutaneously administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of about 300 mg once every 3 weeks. In a certain aspect, provided herein is a method for treating, preventing or managing NAFLD (e.g., alleviating one or more symptoms thereof), the method comprising subcutaneously administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of about 300 mg once every 2 weeks.In a certain aspect, provided herein is a method for treating, preventing or managing NAFLD (e.g., alleviating one or more symptoms thereof), the method comprising subcutaneously administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of about 300 mg once a week. In a certain aspect, provided herein is a method for treating, preventing or managing NAFLD (e.g., alleviating one or more symptoms thereof), the method comprising subcutaneously administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose in the range of about 200 mg to 250 mg once every 3 weeks or once every 2 weeks. In a specific aspect, the method provided herein is used to treat a human subject with hypercholesterolemia (e.g., primary hypercholesterolemia) or dyslipidemia (e.g., mixed dyslipidemia). In a specific aspect, the method provided herein is used to treat a human subject with hypertriglyceridemia (particularly severe hypertriglyceridemia). In certain aspects, the methods provided herein are used to treat a human subject suffering from type 2 diabetes or an obese human subject. In specific aspects, the methods provided herein are used to treat a human subject aged 18 to 55 years, and before administering human FGF21 protein variants, the human subject has (i) 30-45 kg / m. 2 BMI within the range of (including the end value) (wherein for Asian subjects, race-adjusted greater than or equal to 27.5 kg / m 2 BMI), and (ii) triglyceride levels in the range of 150 to 500 mg / dL (1.69-5.65 mmol / L).

[0106] In a specific aspect, provided herein is a method for reducing fat or lipid in the liver, the method comprising administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof (e.g., a subject with NASH or NAFLD) at a dose in the range of 100 mg to 600 mg (or more specifically, at a dose in the range of 200 mg to 400 mg or 100 mg to 300 mg), for example once a day, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month). In a specific aspect, provided herein is a method for reducing fat or lipid in the liver, the method comprising administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof (e.g., a subject with NASH or NAFLD) at a dose of about 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg, for example, once a day, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month). In a certain aspect, provided herein is a method for reducing fat or lipid in the liver, the method comprising administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) subcutaneously to a subject in need thereof (e.g., a subject with NASH or NAFLD) at a dose of about 300 mg once every 4 weeks (or once a month). In a certain aspect, provided herein is a method for reducing fat or lipid in the liver, the method comprising administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) subcutaneously to a subject in need thereof (e.g., a subject with NASH or NAFLD) at a dose of about 300 mg once every 3 weeks. In a certain aspect, provided herein is a method for reducing fat or lipid in the liver, the method comprising administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) subcutaneously to a subject in need thereof (e.g., a subject with NASH or NAFLD) at a dose of about 300 mg once every 2 weeks.In a certain aspect, provided herein is a method for reducing fat or lipid in the liver, the method comprising administering human FGF21 protein variant (e.g., Fc fusion protein, such as V103 (SEQ ID NO: 11)) subcutaneously to a subject in need thereof (e.g., a subject with NASH or NAFLD) at a dose of about 300 mg once a week. In a certain aspect, provided herein is a method for reducing fat or lipid in the liver, the method comprising administering human FGF21 protein variant (e.g., Fc fusion protein, such as V103 (SEQ ID NO: 11)) subcutaneously to a subject in need thereof (e.g., a subject with NASH or NAFLD) at a dose in the range of about 200 mg to 250 mg once every 3 weeks or once every 2 weeks. In a specific aspect, the method provided herein is used to treat a human subject with NASH or NAFLD. In a specific aspect, the method provided herein is used to treat a human subject with hypercholesterolemia (e.g., primary hypercholesterolemia) or dyslipidemia (e.g., mixed dyslipidemia). In certain aspects, provided herein is a method for treating a human subject suffering from hypertriglyceridemia (particularly severe hypertriglyceridemia). In certain aspects, provided herein is a method for treating a human subject suffering from type 2 diabetes or an obese human subject. In certain aspects, provided herein is a method for treating a human subject aged 18 to 55 years, and when screening before applying human FGF21 protein variants, the human subject has (i) at 30-45kg / m. 2 BMI within the range of (including the end value) (wherein for Asian subjects, race-adjusted greater than or equal to 27.5 kg / m 2 BMI), and (ii) triglyceride levels in the range of 150 to 500 mg / dL (1.69-5.65 mmol / L).

[0107] In a particular aspect, provided herein is a method for reducing elevated LDL cholesterol (LDL-C), total cholesterol (total C), triglycerides and / or Apo B, the method comprising administering to a subject in need thereof a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) at a dose in the range of 100 mg to 600 mg, for example, once a day, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month). In a particular aspect, provided herein is a method for reducing elevated LDL-C, total C, triglycerides and / or Apo B, the method comprising administering to a subject in need thereof a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) at a dose in the range of 100 mg to 600 mg (or more specifically, at a dose in the range of 200 mg to 400 mg or 100 mg to 300 mg). In a specific aspect, provided herein is a method for lowering elevated LDL-C, total C, triglycerides and / or Apo B, the method comprising administering to a subject in need thereof a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) at a dose of about 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg, for example, once a day, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month). In a certain aspect, provided herein is a method for reducing elevated LDL-C, total C, triglycerides and / or Apo B, the method comprising administering human FGF21 protein variants (e.g., Fc fusion proteins, such as V103 (SEQ ID NO: 11)) subcutaneously to a subject in need thereof at a dose of about 300 mg once every 4 weeks (once a month). In a certain aspect, provided herein is a method for reducing elevated LDL-C, total C, triglycerides and / or Apo B, the method comprising administering human FGF21 protein variants (e.g., Fc fusion proteins, such as V103 (SEQ ID NO: 11)) subcutaneously to a subject in need thereof at a dose of about 300 mg once every 3 weeks.In a certain aspect, provided herein is a method for reducing elevated LDL-C, total C, triglycerides and / or Apo B, the method comprising administering human FGF21 protein variants (e.g., Fc fusion proteins, such as V103 (SEQ ID NO: 11)) subcutaneously to a subject in need thereof at a dose of about 300 mg once every 2 weeks. In a certain aspect, provided herein is a method for reducing elevated LDL-C, total C, triglycerides and / or Apo B, the method comprising administering human FGF21 protein variants (e.g., Fc fusion proteins, such as V103 (SEQ ID NO: 11)) subcutaneously to a subject in need thereof at a dose of about 300 mg once a week. In a certain aspect, provided herein is a method for reducing elevated LDL-C, total C, triglycerides and / or Apo B, the method comprising administering human FGF21 protein variants (e.g., Fc fusion proteins, such as V103 (SEQ ID NO: 11)) to a subject in need thereof subcutaneously at a dose in the range of about 200 mg to 250 mg once every 3 weeks or once every 2 weeks. In a specific aspect, the method provided herein reduces elevated LDL-C, total C, triglycerides and / or Apo B by at least about 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, or 90% or more. In certain aspects, the method provided herein is used to treat a human subject with hypercholesterolemia (e.g., primary hypercholesterolemia) or dyslipidemia (e.g., mixed dyslipidemia). In certain aspects, the method provided herein is used to treat a human subject with hypertriglyceridemia (particularly severe hypertriglyceridemia). In certain aspects, the method provided herein is used to treat a human subject with type 2 diabetes or an obese human subject. In certain aspects, the method provided herein is used to treat a human subject aged 18 to 55 years, and when screening before applying human FGF21 protein variants, the human subject has (i) at 30-45kg / m. 2 BMI within the range of (including the end value) (wherein for Asian subjects, race-adjusted greater than or equal to 27.5 kg / m 2 BMI), and (ii) triglyceride levels in the range of 150 to 500 mg / dL (1.69-5.65 mmol / L).

[0108] In a specific aspect, provided herein is a method for increasing HDL-C (e.g., increasing HDL-C by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%), the method comprising administering to a subject in need thereof a therapeutically effective amount of a fusion protein comprising a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) at a dose in the range of 100 mg to 600 mg, for example once a day, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month). In a particular aspect, provided herein is a method for increasing HDL-C (e.g., increasing HDL-C by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%), the method comprising administering to a subject in need thereof a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) at a dose in the range of about 200 mg to 400 mg. In one specific aspect, provided herein is a method of increasing HDL-C (e.g., increasing HDL-C by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%), the method comprising administering about 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, The human FGF21 protein variant (e.g., Fc fusion protein, such as V103 (SEQ ID NO: 11)) is administered to a subject in need thereof at a dosage of about 300 mg, 320 mg, 330 mg, 340 mg, or 350 mg, for example, once a day, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month). In a certain aspect, the present invention provides a method for increasing HDL-C (e.g., increasing HDL-C by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%), the method comprising subcutaneously administering a human FGF21 protein variant (e.g., Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dosage of about 300 mg once every 4 weeks (or once a month) or once every 3 weeks or once every 2 weeks.In a certain aspect, provided herein is a method of increasing HDL-C (e.g., increasing HDL-C by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%), the method comprising administering subcutaneously to a subject in need thereof a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) at a dose in the range of about 200 mg to 250 mg once every 3 weeks or once every 2 weeks. In specific aspects, the methods provided herein are used to treat human subjects with hypercholesterolemia (e.g., primary hypercholesterolemia) or dyslipidemia (e.g., mixed dyslipidemia). In specific aspects, the methods provided herein are used to treat human subjects with hypertriglyceridemia (particularly severe hypertriglyceridemia). In certain aspects, the methods provided herein are used to treat human subjects with type 2 diabetes or obese human subjects. In certain aspects, the methods provided herein are used to treat human subjects aged 18 to 55 years who, when screened prior to administration of the human FGF21 protein variant, have (i) a body weight of 30-45 kg / m. 2 BMI within the range of (including the end value) (wherein for Asian subjects, race-adjusted greater than or equal to 27.5 kg / m 2 BMI), and (ii) triglyceride levels in the range of 150 to 500 mg / dL (1.69-5.65 mmol / L).

[0109] In a specific aspect, provided herein is a method for reducing triglyceride levels (e.g., fasting triglyceride levels) (e.g., reducing blood / serum triglyceride levels by at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% or more), the method comprising administering to a subject in need thereof a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) at a dose in the range of 100 mg to 600 mg, for example once a day, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month). In a particular aspect, provided herein is a method for lowering triglyceride levels (e.g., lowering blood / serum triglyceride levels by at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% or more), the method comprising administering to a subject in need thereof a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) at a dose in the range of about 200 mg to 400 mg. In a specific aspect, provided herein is a method of reducing triglyceride levels (e.g., fasting triglyceride levels) (e.g., reducing blood / serum triglyceride levels by at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% or more), the method comprising administering about 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, 350 mg, 360 mg, 370 mg, 380 mg, 390 mg, 400 mg, 410 mg, 420 mg, 430 mg, 440 mg, 450 mg, 460 mg, 470 mg, 480 mg, 490 mg, 500 mg, 510 mg, The human FGF21 protein variant (e.g., Fc fusion protein, such as V103 (SEQ ID NO: 11)) is administered to a subject in need thereof at a dosage of, for example, once a day, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month). In a certain aspect, provided herein is a method for reducing triglyceride levels (e.g., fasting triglyceride levels) (e.g., reducing blood / serum triglyceride levels by at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% or more), the method comprising subcutaneously administering a human FGF21 protein variant (e.g., Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dosage of about 300 mg once every 4 weeks (or once a month) or once every 3 weeks or once every 2 weeks.In a certain aspect, provided herein is a method for reducing triglyceride levels (e.g., fasting triglyceride levels) (e.g., reducing blood / serum triglyceride levels by at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% or more), the method comprising administering human FGF21 protein variants (e.g., Fc fusion proteins, such as V103 (SEQ ID NO: 11)) subcutaneously to a subject in need thereof at a dose in the range of about 200 mg to 250 mg once every 3 weeks or once every 2 weeks or once a week. In a specific aspect, the methods provided herein are capable of reducing triglyceride levels (e.g., fasting triglyceride levels) by at least about 40% or at least about 50% for a period of at least 1-4 weeks. In one aspect, provided herein is a method for reducing triglyceride levels (e.g., fasting triglyceride levels) by at least about 40% to 60% (e.g., at least about 50%) for a period of at least 1-4 weeks, the method comprising subcutaneously administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of about 300 mg once every 4 weeks (or once a month) or once every 3 weeks or once every 2 weeks. In a certain aspect, provided herein is a method of reducing triglyceride levels (e.g., fasting triglyceride levels) by at least about 40% to 60% (e.g., at least about 50%) for at least 1-4 weeks, the method comprising administering a human FGF21 protein variant (e.g., Fc fusion protein, such as V103 (SEQ ID NO: 11)) subcutaneously to a subject in need thereof at a dose ranging from 200 mg to 300 mg (e.g., at least 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, or 280 mg) once every 4 weeks (or once a month) or once every 3 weeks or once every 2 weeks. In a specific aspect, the method provided herein is used to treat a human subject with hypercholesterolemia (e.g., primary hypercholesterolemia) or dyslipidemia (e.g., mixed dyslipidemia). In a specific aspect, the method provided herein is used to treat a human subject with hypertriglyceridemia (particularly severe hypertriglyceridemia). In certain aspects, the method provided herein is used to treat a human subject suffering from type 2 diabetes or an obese human subject. In a specific aspect, the method provided herein is used to treat a human subject aged 18 to 55 years old, and when screening before using human FGF21 protein variants, the human subject has (i) at 30-45kg / m. 2 BMI within the range of (including the end value) (wherein for Asian subjects, race-adjusted greater than or equal to 27.5 kg / m 2 BMI), and (ii) triglyceride levels in the range of 150 to 500 mg / dL (1.69-5.65 mmol / L).

[0110] In one specific aspect, the method provided herein is capable of reducing triglyceride levels (e.g., fasting triglyceride levels) by at least about 40% or at least about 50%, for example, for a period of at least 1-4 weeks, the method comprising subcutaneously administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose in the range of approximately 250 mg to 300 mg (e.g., 300 mg) once every 4 weeks (or once a month) or once every 3 weeks. In one aspect, provided herein is a method for reducing triglyceride levels (e.g., fasting triglyceride levels) by at least about 40% to 60% (e.g., at least about 50%), for example for a period of at least 1-4 weeks, the method comprising subcutaneously administering a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose ranging from about 250 mg to 300 mg (e.g., 300 mg) once every 4 weeks (or once a month), once every 3 weeks, or once every 2 weeks.

[0111] In a specific aspect, provided herein is a method for reducing cardiovascular risk (e.g., reducing it by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% or more), the method comprising administering a therapeutically effective amount of a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose ranging from 100 mg to 600 mg, for example once a day, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month). In a particular aspect, provided herein is a method for reducing cardiovascular risk (e.g., reducing it by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% or more), the method comprising administering to a subject in need thereof a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) at a dose in the range of about 200 mg to 400 mg or 100 mg to 300 mg, for example once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks. In one specific aspect, provided herein is a method of reducing cardiovascular risk (e.g., reducing by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% or more), the method comprising taking about 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, The human FGF21 protein variant (e.g., Fc fusion protein, such as V103 (SEQ ID NO: 11)) is administered to a subject in need thereof at a dosage of, for example, once a day, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month). In a certain aspect, provided herein is a method for reducing cardiovascular risk (e.g., reducing at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% or more), the method comprising subcutaneously administering a human FGF21 protein variant (e.g., Fc fusion protein, such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dosage of about 300 mg once every 4 weeks (or once a month).In a certain aspect, provided herein is a method for reducing cardiovascular risk (e.g., reducing at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% or more), the method comprising administering human FGF21 protein variant (e.g., Fc fusion protein, such as V103 (SEQ ID NO: 11)) subcutaneously to a subject in need thereof at a dose of about 300 mg once every 3 weeks. In a certain aspect, provided herein is a method for reducing cardiovascular risk (e.g., reducing at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%), the method comprising administering human FGF21 protein variant (e.g., Fc fusion protein, such as V103 (SEQ ID NO: 11)) subcutaneously to a subject in need thereof at a dose in the range of about 200 mg to 250 mg once every 3 weeks. In a certain aspect, provided herein is a method for reducing cardiovascular risk (e.g., reducing at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% or more), the method comprising administering subcutaneously to a subject in need thereof a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)) at a dose of about 300 mg once every 2 weeks or once a week. In specific aspects, the methods provided herein are used to treat human subjects with hypercholesterolemia (e.g., primary hypercholesterolemia) or dyslipidemia (e.g., mixed dyslipidemia). In specific aspects, provided herein are methods for treating human subjects with hypertriglyceridemia (particularly severe hypertriglyceridemia). In certain aspects, provided herein are methods for treating human subjects with type 2 diabetes or obese human subjects. In a specific aspect, the methods provided herein are used to treat a human subject aged 18 to 55 years, who prior to administration of the human FGF21 protein variant has (i) a body weight of 30-45 kg / m. 2 BMI within the range of (including the end value) (wherein for Asian subjects, race-adjusted greater than or equal to 27.5 kg / m 2 BMI), and (ii) triglyceride levels in the range of 150 to 500 mg / dL (1.69-5.65 mmol / L).

[0112] In certain aspects, the methods provided herein are used to treat human subjects aged 18 to 55 years who, when screened prior to administration of a human FGF21 protein variant, have (i) a body weight of 30-45 kg / m 2 BMI within the range of (including the end value) (wherein for Asian subjects, race-adjusted greater than or equal to 27.5 kg / m 2BMI), and (ii) triglyceride levels in the range of 150 to 500 mg / dL (1.69-5.65 mmol / L).

[0113] In various aspects, the methods provided herein are used to treat human subjects who are at least 18 years old. In other aspects, the methods provided herein are used to treat human subjects who are at least 55 years old. In other aspects, the methods provided herein are used to treat human subjects who are at least 60 years old. In other aspects, the methods provided herein are used to treat human subjects who are at least 65 years old.

[0114] In certain aspects, the methods provided herein are used to treat patients with 2 In certain aspects, the methods provided herein are used to treat subjects with a BMI greater than or equal to 25 kg / m 2 In certain aspects, the methods provided herein are used to treat a subject having a BMI greater than or equal to 30 kg / m 2 In certain aspects, the methods provided herein are used to treat a subject having a BMI of greater than or equal to 35 kg / m 2 In some aspects, the methods provided herein are used to treat a human subject with a BMI of ≥ 27.5 for Asian individuals (with adjustment for race).

[0115] In a particular aspect, the method provided herein is used to treat a human subject with a triglyceride level in the range of 150-500mg / dL (1.69-5.65mmol / L) before applying human FGF21 protein variants. In a particular aspect, the method provided herein is used to treat a human subject with a triglyceride level of at least 150mg / dL (at least 1.69mmol / L) before applying human FGF21 protein variants. In a particular aspect, the method provided herein is used to treat a human subject with a triglyceride level of at least 200mg / dL when screening before applying human FGF21 protein variants. In a particular aspect, the method provided herein is used to treat a human subject with a triglyceride level of at least 500mg / dL (at least 5.65mmol / L) before applying human FGF21 protein variants.

[0116] In certain aspects, a human subject having one or more of the following is not treated according to the methods provided herein:

[0117] History of hepatobiliary disease, cholelithiasis, or cholestasis (on medical history or ultrasound screening), hepatic encephalopathy, esophageal varices, or portal vein shunts;

[0118] Liver disease or liver damage as indicated by liver function tests (ALT, AST, GGT, alkaline phosphatase, or serum bilirubin) above the upper limit of normal at screening or baseline;

[0119] Chronic infection with human immunodeficiency virus (HIV), hepatitis B (HBV) or hepatitis C (HCV);

[0120] A positive HBV surface antigen (HBsAg) test or, under standard local practice, a positive HBV core antigen test.

[0121] Positive (detectable) HCV RNA;

[0122] A history of pancreatic damage or pancreatitis or other pancreatic disease;

[0123] Amylase or lipase above ULN at screening or baseline;

[0124] History of hypersensitivity reaction to similar biologics, FGF21 protein analogs, or Fc fusion proteins;

[0125] A history of bone disorders (including but not limited to osteoporosis, osteopenia, osteomalacia, severe vitamin D deficiency); and

[0126] Plasma 25-hydroxyvitamin D level below the lower limit of the normal range at screening.

[0127] In one aspect, provided herein is a combination therapy for treating, preventing, or managing a metabolic disorder or a cardiovascular disorder, comprising administering a therapeutically effective amount of an FGF21 protein variant described herein (e.g., an Fc fusion protein, such as V103) and one or more additional therapeutically active agents (e.g., therapeutic agents for a metabolic disorder or a cardiovascular disorder). Non-limiting examples of other therapeutically active agents for use in combination with the FGF21 protein variants provided herein include obesity therapeutics (e.g., phentermine / topiramate, orlistat, lorcaserin, liraglutide, bupropion / naltrexone), hypertension therapeutics (e.g., diuretics, β-blockers, α-blockers, ACE inhibitors, angiotensin II receptor blockers (ARBs), direct renin inhibitors, calcium channel blockers, central agonists, peripheral adrenergic blockers, vasodilators, and combinations), diabetes therapeutics (e.g., insulin, α-glucosidase inhibitors, biguanides, dopamine agonists, DPP-4 inhibitors, glucagon-like peptides, meglitinides, sodium glucose transporter (SGLT) inhibitors, sulfonylureas, thiazolidinediones, pramlintide), NAFLD / NASH and cardiovascular therapeutics (e.g., statins, fibrates, aspirin, anticoagulants).

[0128] In one aspect, provided herein is a combination therapy for treating, preventing or managing a metabolic disorder or a cardiovascular disorder, the combination therapy comprising administering a therapeutically effective amount of a FGF21 protein variant described herein (e.g., an Fc fusion protein, such as V103) and one or more therapeutically active agents selected from the group consisting of amiloride (Midamor), bumetanide (Bumex), chlorthalidone (Hygroton), chlorothiazide (Diuril), furosemide (Lasix), furosemide (Lasix), furosemide (Lasix), furosemide (Lasix), furosemide (Lasix), furosemide (Lasix), furosemide (Lasix), furosemide (Lasix), furosemide (Lasix), furosemide (Lasix), furosemide (Lasix), furosemide (Lasix), furosemide (Lasix), furosemide (Lasix), furosemide (Lasix), furosemide (Lasix), furosemide (Lasix), furosemide (Lasix), furosemide (Lasix), furosemide (Lasix), furosemide (Lasix), furosemide (Lasix), furosemide (Lasix), chlorthalidone (chlorthalidone) (Hygroton), chlorothiazide (Diuril ... ), hydrochlorothiazide or HCTZ (Esidrix, Hydrodiuril, Microzide), indapamide (Lozol), metolazone (Mykrox, Zaroxolyn), spironolactone (Aldactone), triamterene (Dyrenium), acebutolol (Sectra1), amoxicillin (Acebutolol), Atenolol (Tenormin), Betaxolol (Kerlone), Bisoprolol (Zebeta), Carteolol (Cartrol), Metoprolol (Lopressor, Toprolol XL), Nadolol (Corgard), Nebivolol (Bysto1ic), Penbutolol (Levatol), o1), Pindolol (Visken), Proprano1o1 (Inderal), Sotalol (Betapace), Timo1o1 (Timolol (Biocadren)), Doxazosin (Cardura), Prazosin (Minipress), Terazosin (Hytrin), Benazepril (Lotensin),Captopril (Capoten), Enalapril (Vasotec), Fosinopril (Monopril), Lisinopril (Prinivil, Zestril), Moexipril (Univasc), Perindopril (Aceon), Quinapril (Accupril), Ramipril (A1ta ce), Trandolapril (Mavik), Norvasc (amlodipine), P1endil (felodipine), DynaCirc (isradipine), Cardene (nicardipine), Procardia XL, Adalat (nifedipine), Cardizem, Dilacor, Tiazac, Diltia XL (diltiazem), Sular (nisoldipine), Isoptin, Calan, Verelan, Covera-HS (verapamil), Capoten (captopril), Vasotec (enalapril), Prinivil, Zestril (lisinopril), Lotensin (benazepril), zepril), Monopril (fosinopril), A1tace (ramipril), Accupril (quinapril), Aceon (perindopril), Mavik (trandolapril), Univasc (moexipril), Atacand (candesartan), Avapro (irbesartan),Benicar (olmesartan), Cozaar (losartan), Diovan (valsartan), Micardis (telmisartan), Teveten (eprosartan), chlorthalidone (Hygroton), chlorothiazide (Diuril), hydrochlorothiazide, or HCTZ (Es idrix, Hydrodiuril, Microzide, indapamide (Lozol), metolazone (Mykrox, Zaroxolyn), amiloride (Midamor), bumetanide (Bumex), furosemide (furosemide (Lasix)), spironolactone (Aldactone), triamterene (Benzotrene), and spironolactone (Aldactone). Dyrenium), Acebutolol (Sectral), Atenolol (Tenormin), Betaxolol (Kerlone), Bisoprolol (Zebeta, Ziac), Carteolol (Cartrol), Carvedilol (Coreg), Labetalol (Normodyne, Trandate), Metoprolol (Metoprolol), rolol)(Lopressor, Toprol-XL), Nadolol(Corgard), Nebivolol(Bysto1ic), Penbutolol(Levatol), Pindolol(Visken), Propanolol(Inderal), Sotalol(Betapace), Timo1(Blocadren),Fibric acid derivatives, hydrochloric acid, and omega-3 fatty acids, fenofibrate, gemfibrozil, atorvastatin, fluvastatin, lovastatin, pitavastatin, pravastatin, rosuvastatin, simvastatin, pramlintide, acarbose (Precose), miglitol (Glyset), metformin, bromocriptine, alogliptin, linagliptin, saxagliptin, sitagliptin, albiglutide (Tanzeum), dulaglutide (Trulicity), exenatide (Byetta) ), exenatide extended release (Bydureon), liraglutide (Victoza), nateglinide (Starlix), repaglinide (Prandin), repaglinide-metformin (Prandimet), dapagliflozin (Farxiga), repaglinide-metformin (Xigduo XR), canagliflozin (Invokana), canagliflozin-metformin (Invokamet), empagliflozin (Jardiance), empagliflozin-linagliptin (G1yxambi), empagliflozin-metformin (Synjardy), sotagliflozin, tofogliflozin, remogliflozin, luseogliflozin, ipragliflozin, atigliflozin, bexagliflozin, henagliflozin, icogliflozin, glimepiride (Amaryl), glimepiride-pioglitazone (Duetact),Glimeperide-rosiglitazone (Avandaryl), gliclazide, glipizide-metformin (Metaglip), glyburide (DiaBeta, Glynase, Micronase), glyburide-metformin (Glucovance), chlorpropamide (Chlorpropamide) ropamide (Diabinese), tolazamide (Tolinase), tolbutamide (Orinase, Tol-Tab), rosiglitazone (Avandia), rosiglitazone-glimepiride (Avandaryl), rosiglitazone-metformin (Amaryl M), pioglitazone (Actos), pioglitazone-alogliptin (Oseni), pioglitazone-glimepiride (Duetact), and pioglitazone-metformin (Actoplus Met, Actoplus Met XR).

[0129] In some embodiments, sodium glucose transporter (SGLT) inhibitor is selected from dapagliflozin, empagliflozin, canagliflozin, ertugliflozin, sotagliflozin, tofogliflozin, remogliflozin, 1useogliflozin, ipragliflozin, atigliflozin, bexagliflozin, henagliflozin, licogliflozin and any pharmaceutically acceptable salt thereof. In some embodiments, sodium glucose transporter (SGLT) inhibitor is dapagliflozin. In some embodiments, sodium glucose transporter (SGLT) inhibitor is empagliflozin. In some embodiments, sodium glucose transporter (SGLT) inhibitor is canagliflozin. In some embodiments, sodium glucose transporter (SGLT) inhibitor is ertugliflozin. In some embodiments, the sodium glucose transporter (SGLT) inhibitor is licogliflozin.

[0130] In some embodiments, the sodium glucose transporter (SGLT) inhibitor is dapagliflozin or a pharmaceutically acceptable salt thereof. In some embodiments, the sodium glucose transporter (SGLT) inhibitor is empagliflozin or a pharmaceutically acceptable salt thereof. In some embodiments, the sodium glucose transporter (SGLT) inhibitor is canagliflozin or a pharmaceutically acceptable salt thereof. In some embodiments, the sodium glucose transporter (SGLT) inhibitor is ergagliflozin or a pharmaceutically acceptable salt thereof. In some embodiments, the sodium glucose transporter (SGLT) inhibitor is licogliflozin or a pharmaceutically acceptable salt thereof.

[0131] In certain aspects, the methods provided herein comprising administering a FGF21 protein variant (e.g., an Fc fusion protein such as V103) are used as an adjunct to diet (e.g., a healthy diet, a calorie-restricted diet), exercise, and / or other lifestyle changes.

[0132] FGF21 variants

[0133] Modifications to FGF21 can improve half-life and / or potency compared to wild-type FGF21 to provide FGF21 therapeutics for treating or managing metabolic or cardiovascular disorders (e.g., alleviating one or more symptoms of the disorder) or for reducing cardiovascular risk with clinically preferred dosing regimens.

[0134] In certain aspects, provided herein are FGF21 protein variants (e.g., the FGF21 protein variants described in Table 1) for use in a method of treating or managing a metabolic disorder or a cardiovascular disorder (e.g., alleviating one or more symptoms of the disorder) or for reducing cardiovascular risk.

[0135] The acceptable amino acid substitutions and modifications constituting the differences between the FGF21 polypeptides and protein variants and mutants and wild-type FGF21 of the methods provided herein include, but are not limited to, one or more amino acid substitutions (including substitutions with non-naturally occurring amino acid analogs) and truncations. Therefore, FGF21 protein variants include, but are not limited to, site-directed FGF21 mutants, truncated FGF21 polypeptides, proteolytically resistant FGF21 mutants, reduced aggregated FGF21 mutants, FGF21 combination mutants, FGF21 conjugates (e.g., fatty acid-FGF21 conjugates, PEG-FGF21 conjugates) and FGF21 fusion proteins (e.g., Fc domain fusion proteins, human serum albumin fusion proteins).

[0136] In one aspect, the FGF21 protein variant comprises substitutions relative to wild-type FGF21 at one or more or all of the following residues (according to the numbering scheme of SEQ ID NO: 1): Q55, R105, G148, K150, P158, S195, P199, and G202.

[0137] In one aspect, the FGF21 protein variant comprises one or more or all of the following substitutions relative to wild-type FGF21 (according to the numbering scheme of SEQ ID NO: 1): Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A.

[0138] Exemplary fusion protein sequences provided herein are listed in Table 1. The description of the fusion includes the FGF21 variant and linker (when applicable). The changes or substitutions adopted by the FGF21 variant are numbered and described relative to wild-type FGF21.

[0139] For example, "Variant 101 (V101)" (SEQ ID NO: 10) is an Fc-FGF21 fusion with two amino acid linkers and the following substitutions relative to wild-type FGF21 (according to the numbering scheme of SEQ ID NO: 1): Q55C, A109T, G148C, K150R, P158S, P174L, S195A, P199G, and G202A.

[0140] By way of another example, "Variant 103 (V103)" (SEQ ID NO: 11) is an Fc-FGF21 fusion having two amino acid linkers (GS) and the following substitutions relative to wild-type FGF21 (according to the numbering scheme of SEQ ID NO: 1): Q55C, R105K, G148C, K150R, P158S, S195A, P199G, G202A.

[0141] Table 1: Exemplary FGF21 variants

[0142]

[0143]

[0144] *-Note that unless otherwise stated, the FGF21 wild-type sequence in this table refers to NCBI reference sequence number NP061986.1 (SEv ID NO: 1).

[0145] All mutations in the FGF21 portion and the corresponding amino acid numbering of said mutations refer to (SEQ ID NO: 1) instead of this table which may also include the full length sequence of the Fc and linker regions.

[0146] In a specific aspect, provided herein is a method for treating, preventing or managing a metabolic disorder or a cardiovascular disorder (e.g., obesity, NALFD, NASH or diabetes), the method comprising administering a human FGF21 protein variant, which is a fusion protein comprising a human Fc region (e.g., a modified human Fc region) fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G and G202A numbered according to SEQ ID NO: 1.

[0147] In a specific aspect, provided herein is a method for treating, preventing or managing a metabolic disorder or a cardiovascular disorder (e.g., obesity, NALFD, NASH or diabetes), the method comprising administering a human FGF21 protein variant, which is a fusion protein comprising a human Fc region (e.g., a modified human Fc region) fused to a mature human FGF21 protein or a fragment thereof, such as V103 comprising the amino acid sequence of SEQ ID NO: 11.

[0148] Functional assays for determining or measuring the biological activity of FGF21 or its variants or fragments thereof have been described, for example, see PCT Publication No. WO 2013 / 049247, which is incorporated herein by reference in its entirety. In particular aspects, the functional assay is an in vitro assay. In other aspects, the functional assay is an in vivo assay. In certain aspects, the functional assay is an ex vivo assay.

[0149] In certain aspects, FGF21 downstream biomarkers can be assessed to determine the biological activity of FGF21 or its variant or its fragment. As used herein, "FGF21 downstream biomarkers" are genes or gene products, or measurable markers of genes or gene products. In some aspects, genes or activity as downstream markers of FGF21 show the expression level of change, or show in vascular tissue. In some aspects, the activity of downstream markers in the presence of FGF21 regulators changes. In some aspects, when FGF21 is disturbed by FGF21 regulators, downstream markers show the expression level of change. FGF21 downstream markers include but are not limited to glucose or 2-deoxy-glucose uptake, pERK and other phosphorylation or acetylated proteins or NAD levels.

[0150] Pharmaceutical compositions and formulations

[0151] In a particular aspect, provided herein is a pharmaceutical composition comprising an FGF21 protein variant (e.g., an FGF21 protein variant described in Table 1, e.g., V103 (SEQ ID NO: 11)) for use in treating, preventing and / or managing metabolic disorders or cardiovascular disorders (e.g., hypertriglyceridemia and risk of heart disease, insulin resistance (e.g., patients with genetic mutations of the insulin receptor) and lipodystrophy, diabetes, obesity and non-alcoholic fatty liver disease (NAFLD) / non-alcoholic steatohepatitis (NASH)).

[0152] The pharmaceutical compositions described herein may include a therapeutically effective amount of a FGF21 protein variant and a pharmaceutically or physiologically acceptable carrier. The carrier is typically selected to be suitable for the intended mode of administration, and may include agents for changing, maintaining or maintaining, for example, the pH, osmotic pressure, viscosity, clarity, color, isotonicity, odor, sterility, stability, dissolution or release rate, adsorption and / or penetration of the composition. Typically, these carriers include aqueous or alcohol / aqueous solutions, emulsions or suspensions, including saline and / or buffered media.

[0153] Suitable agents for inclusion in the pharmaceutical composition include, but are not limited to, amino acids (e.g., glycine, glutamine, asparagine, arginine, histidine or lysine), antimicrobials, antioxidants (e.g., ascorbic acid, sodium sulfite or sodium bisulfite), buffers (e.g., borates, bicarbonates, Tris-HCl, citrates, phosphates or other organic acids), bulking agents (e.g., mannitol or glycine), chelating agents (e.g., ethylenediaminetetraacetic acid (EDTA)), complexing agents (e.g., caffeine, polyvinylpyrrolidone, β-cyclodextrin or hydroxypropyl-β-cyclodextrin), bulking agents, monosaccharides, disaccharides and other carbohydrates (e.g., glucose, mannose or dextrin), proteins (e.g., free serum albumin, gelatin, or immunoglobulins), colorants, flavorings and diluents, emulsifiers, hydrophilic polymers (e.g., polyvinylpyrrolidone), low molecular weight The invention also includes an aqueous solution of at least one amino acid polypeptide, a salt-forming counterion (e.g., sodium), a preservative (e.g., benzalkonium chloride, benzoic acid, salicylic acid, thimerosal, phenylethyl alcohol, methylparaben, propylparaben, chlorhexidine, sorbic acid, or hydrogen peroxide), a solvent (e.g., glycerol, propylene glycol, or polyethylene glycol), a sugar alcohol (e.g., mannitol or sorbitol), a suspending agent, a surfactant or wetting agent (e.g., pluronics; PEG; sorbitan esters; polysorbates (e.g., polysorbate 20 or polysorbate 80); Triton; tromethamine; lecithin; cholesterol, or tyloxapal), a stability enhancer (e.g., sucrose or sorbitol), a tonicity enhancer (e.g., an alkali metal halide (e.g., sodium chloride or potassium chloride), or mannitol sorbitol), a delivery vehicle, a diluent, an excipient, and / or a pharmaceutical adjuvant.

[0154] Parenteral vehicles include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride and lactated Ringer's solution. Suitable physiologically acceptable thickeners may be included, such as carboxymethylcellulose, polyvinylpyrrolidone, gelatin and alginate. Intravenous vehicles include liquid and nutrient supplements and electrolyte supplements, such as those based on Ringer's dextrose. In some cases, it is preferred to include an agent that adjusts the tension of the composition, for example, sugars, polyols such as mannitol, sorbitol or sodium chloride in the pharmaceutical composition. For example, in many cases, it is desirable that the composition is substantially isotonic. Preservatives and other additives, such as antimicrobials, antioxidants, chelating agents and inert gases may also be present. The exact formulation will depend on the route of administration. Other related principles, methods and components of pharmaceutical formulations are well known. (See, for example, Allen, Loyd V., ed., (2012) Remington's Pharmaceutical Sciences, 22nd edition, which is incorporated by reference for this purpose)

[0155] When parenteral administration is contemplated, the pharmaceutical composition is generally in the form of a sterile, pyrogen-free, parenterally acceptable composition. Particularly suitable vehicles for parenteral injection are properly preserved sterile isotonic solutions. The pharmaceutical composition may be in the form of a lyophilisate, such as a lyophilized cake.

[0156] In certain aspects, the pharmaceutical composition provided herein (e.g., a composition comprising FGF21 protein variant V103) is used for subcutaneous administration. Suitable formulation components and methods for subcutaneous administration of polypeptide therapeutic agents (e.g., antibodies, fusion proteins, etc.) are known in the art. See, for example, disclosed U.S. Patent Application No. 2011 / 0044977 and U.S. Patent No. 8,465,739 and U.S. Patent No. 8,476,239, each of which is incorporated by reference for this purpose. Typically, the pharmaceutical composition for subcutaneous administration contains a suitable stabilizer (e.g., amino acid, such as methionine and or sugar, such as sucrose), a buffer and / or a tension agent.

[0157] In a specific aspect, provided herein is a pharmaceutical composition as a lyophilisate comprising an FGF21 protein variant (e.g., an FGF21 protein variant shown in Table 1, such as V103). In a specific aspect, a pharmaceutical composition provided herein as a lyophilized formulation comprising an FGF21 protein variant (e.g., an FGF21 protein variant shown in Table 1, such as V103) is reconstituted into a solution prior to administration to a subject (e.g., subcutaneous administration).

[0158] In certain aspects, the pharmaceutical composition provided herein comprising FGF21 protein variants (e.g., the FGF21 protein variants shown in Table 1, such as V103) is in the form of a solution (e.g., after lyophilisate reconstruction) of FGF21 protein variants (e.g., V103) comprising 100 mg / mL. In some aspects, the pharmaceutical composition provided herein comprising FGF21 protein variants (e.g., the FGF21 protein variants shown in Table 1, such as V103) is in the form of a solution (e.g., after lyophilisate reconstruction) of FGF21 protein variants (e.g., V103) comprising at least about 50 mg / mL, 60 mg / mL, 70 mg / mL, 80 mg / mL, 90 mg / mL, 100 mg / mL, 110 mg / mL, 120 mg / mL, 130 mg / mL, 140 mg / mL or 150 mg / mL. In some aspects, the pharmaceutical composition provided herein comprising FGF21 protein variants (e.g., FGF21 protein variants shown in Table 1, such as V103) is in the form of a solution (e.g., after lyophilisate reconstitution) of about 50 mg / mL to 150 mg / mL or 100 mg / mL to 150 mg / mL of FGF21 protein variants (e.g., V103). In one aspect, the pharmaceutical composition provided herein comprising FGF21 protein variants (e.g., FGF21 protein variants shown in Table 1, such as V103) is in the form of a solution (e.g., after lyophilisate reconstitution) of about 75 mg / mL to 150 mg / mL of V103 (e.g., 100 mg / mL V103 (SEQ ID NO: 11)).

[0159] In some aspects, the pharmaceutical composition provided herein comprising FGF21 protein variants (e.g., FGF21 protein variants shown in Table 1, such as V103) is in the form of a solution (e.g., after lyophilisate reconstitution) comprising a tromethamine buffer (e.g., a concentration of 10mM, 20mM, 30mM, 40mM, or 50mM tromethamine buffer). In some aspects, the pharmaceutical composition provided herein comprising FGF21 protein variants (e.g., FGF21 protein variants shown in Table 1, such as V103) is in the form of a solution (e.g., after lyophilisate reconstitution) comprising a tromethamine buffer (e.g., a concentration of 10mM to 50mM tromethamine buffer, such as 30mM tromethamine buffer).

[0160] In some aspects, the pharmaceutical compositions provided herein comprising FGF21 protein variants (e.g., FGF21 protein variants shown in Table 1, such as V103) are in the form of a solution (e.g., at least about 200mM, 210mM, 220mM, 230mM, 240mM, 250mM, 260mM, 270mM, 280mM, 290mM or 300mM sucrose) comprising sucrose. In some aspects, the pharmaceutical compositions provided herein comprising FGF21 protein variants (e.g., FGF21 protein variants shown in Table 1, such as V103) are in the form of a solution (e.g., at least about 250mM to 300mM sucrose, such as 270mM) comprising sucrose (e.g., at least about 200mM, 210mM, 220mM, 230mM, 240mM, 250mM, 260mM, 270mM, 280mM, 290mM or 300mM sucrose) comprising sucrose.

[0161] In some aspects, the pharmaceutical compositions provided herein comprising FGF21 protein variants (e.g., FGF21 protein variants shown in Table 1, such as V103) are in the form of a solution (e.g., after lyophilisate reconstitution) comprising polysorbate (e.g., polysorbate 20). In particular aspects, the pharmaceutical compositions provided herein comprising FGF21 protein variants (e.g., FGF21 protein variants shown in Table 1, such as V103) are in the form of a solution (e.g., after lyophilisate reconstitution) comprising at least about 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, v.07%, 0.08%, 0.09%, or 0.10% polysorbate 20. In a particular aspect, the pharmaceutical compositions provided herein comprising an FGF21 protein variant (e.g., an FGF21 protein variant shown in Table 1, such as V103) are in the form of a solution (e.g., after reconstitution of a lyophilisate) comprising 0.02% to 0.10% polysorbate 20 (e.g., 0.06% polysorbate 20).

[0162] In some aspects, the pharmaceutical compositions provided herein comprising FGF21 protein variants (e.g., FGF21 protein variants shown in Table 1, such as V103) are in the form of a solution (e.g., a pH of 8.0) in the range of 6.5 to 9 (e.g., after reconstitution of a lyophilisate). In some aspects, the pharmaceutical compositions provided herein comprising FGF21 protein variants (e.g., FGF21 protein variants shown in Table 1, such as V103) are in the form of a solution (e.g., after reconstitution of a lyophilisate) at a pH of at least about 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0 or 9.5.

[0163] In some aspects, the pharmaceutical compositions provided herein comprising an FGF21 protein variant (e.g., an FGF21 protein variant shown in Table 1, such as V103) are in the form of a solution (e.g., after reconstitution of a lyophilisate) comprising tromethamine buffer, sucrose, and polysorbate 20.

[0164] In a particular aspect, the pharmaceutical composition provided herein comprising an FGF21 protein variant (e.g., an FGF21 protein variant shown in Table 1, such as V103) is in the form of a solution (pH 7.5 to 8.5, e.g., pH 8.0) comprising about 80 mg / mL to 100 mg / mL of an FGF21 protein variant (e.g., V103 (SEQ ID NO: 11)), about 30 mM tromethamine buffer, 270 mM sucrose, and 0.06% polysorbate 20 (e.g., after reconstitution of a lyophilisate).

[0165] In a particular aspect, the pharmaceutical composition provided herein comprising an FGF21 protein variant (e.g., an FGF21 protein variant shown in Table 1, such as V103) is in the form of a solution comprising about 100 mg / mL of an FGF21 protein variant (e.g., V103 (SEQ ID NO: 11)), about 30 mM tromethamine buffer, 270 mM sucrose, and 0.06% polysorbate 20 (e.g., after reconstitution of the lyophilisate) (pH 8.0).

[0166] In certain aspects, provided herein is a method for preparing a pharmaceutical composition comprising an FGF21 protein variant (e.g., an FGF21 protein variant shown in Table 1, such as V103), the method comprising formulating the FGF21 protein variant as a lyophilisate suitable for reconstitution into a solution.

[0167] In certain aspects, provided herein is a method for preparing a pharmaceutical composition comprising an FGF21 protein variant (e.g., an FGF21 protein variant shown in Table 1, such as V103), the method comprising reconstituting a lyophilisate into a solution described herein, such as a solution (pH 8.0) comprising up to about 100 mg / mL (e.g., about 50 mg / mL to about 100 mg / mL0) of an FGF21 protein variant (e.g., V103 (SEQ ID NO: 11)), about 30 mM tromethamine buffer, 270 mM sucrose, and 0.06% polysorbate 20.

[0168] In certain aspects, provided herein is a method for preparing a pharmaceutical composition comprising an FGF21 protein variant (e.g., an FGF21 protein variant shown in Table 1, such as V103), the method comprising preparing a solution (pH 8.0) comprising about 100 mg / mL of an FGF21 protein variant (e.g., V103 (SEQ ID NO: 11)), about 30 mM tromethamine buffer, 270 mM sucrose, and v.06% polysorbate 20.

[0169] In one specific aspect, the method for preparing the pharmaceutical composition described herein comprises the step of diluting a solution containing a FGF21 protein variant (e.g., a FGF21 protein variant shown in Table 1, such as V103) to a concentration in the range of 10 mg / mL to 90 mg / mL. In one specific aspect, the solution is diluted with a saline solution (e.g., a 0.9% saline solution).

[0170] V. Examples

[0171] The following examples are intended only to illustrate the present invention and not to limit it. Abbreviations are used as is conventional in the art.

[0172] Example 1

[0173] Fc-FGF21 variant fusion proteins (e.g., V103 comprising SEQ ID NO: 11) have been described, see, e.g., PCT Publication No. WO 2013 / 049247. For example, V103 has been reported to induce 2-deoxyglucose turnover by mouse 3T3L1 adipocytes, to induce ERK signaling in a cell-based ERK phosphorylation assay, and to reduce total plasma glucose, plasma insulin, and body weight in ob / ob mice (mice lacking functional leptin as a mouse model for type 2 diabetes). V103 has been reported to have a longer half-life and to be more thermodynamically stable, e.g., as assessed by melting temperature.

[0174] The PK characteristics of V103 in cynomolgus monkeys have been established in a 2-dose IV and SC injection toxicity study with a 4-week recovery period and in a 13-week IV and SC injection toxicity study with a 13-week recovery period, respectively.

[0175] The pharmacokinetics of V103 were similar in both studies, with no significant gender differences. Moderate accumulation of V103 was observed in the 13-week GLP toxicity study (accumulation index 1.15-2.36 across the four dose groups, as measured by C 最大 and AUC 0-7d).

[0176] In the dose range of 0.3-100 mg / kg, V103 exposure (C 最大 The pharmacokinetics of intravenous administration are biexponential and have a linear terminal elimination phase. V103 is slowly absorbed after subcutaneous administration (100 mg / kg), and maximum exposure is observed 1-4 days after SC administration. The terminal elimination phase is not absorption-limited and the estimated subcutaneous bioavailability is 60%-90%. Table 2 below summarizes the average toxicokinetic parameters calculated for the penultimate (85th day) and final dose (92nd day) of the 13-week GLP compliance toxicology study.

[0177] Table 2 Mean TK parameters obtained at the penultimate (Day 85) and final doses (Day 92) of the 13-week GLP-compliant toxicity study in cynomolgus monkeys

[0178]

[0179] Day 85, n=6 animals / main group, 0.3 mg / kg, 3.0 mg / kg, 30 mg / kg iv and 30 mg / kg sc V103. Day 92, n=4 animals / recovery group, 30 mg / kg iv and 30 mg / kg sc V103.

[0180] C 最大 / dose ((μg / mL) / (mg / kg)); AUC0-7d (μg×h / mL); AUC0-7d / dose ((μg×h / mL) / (mg / kg))

[0181] The GLP-validated MSD-based PK assay used to determine V103 concentrations in cynomolgus monkey serum quantified bioactive but also partially truncated / inactive forms of V103.

[0182] Consistent with the rat pharmacokinetics, HPLC-MS / MS evaluation confirmed that in cynomolgus monkeys, the central portion of the FGF21 subdomain of V103 (T 1 / 2 About 9-12 days) than the C-terminal subdomain (T 1 / 2 Table 3 below summarizes the pharmacokinetic properties of the bioactive and truncated / inactive forms of V103 determined in a 13-week GLP-compliant toxicology study.

[0183] Table 3 In vivo integrity of V103 following weekly IV and SC administration in a 13-week GLP-compliant toxicity study in cynomolgus monkeys

[0184]

[0185] TK parameters were calculated from the average TK profile after the last dose (day 92)

[0186] *AUC0-90 days (μg×h / mL)

[0187] a As measured by MSD-based assay b As measured by LC-MS / MS based assay

[0188] Anti-V103 antibodies (ADA) were observed in many animals. In most cases, the ADA response to treatment was associated with accelerated clearance of V103, however, the overall effect on the total and sustained exposure of V103 was minimal in these studies. The selectivity of the ADA response was evaluated during the 13-week GLP toxicity study, and no ADA-positive samples cross-reacted with endogenous wild-type FGF21.

[0189] Example 2

[0190] The clinical study aims to evaluate the safety and tolerability of subcutaneously administered multiple doses of V103 over 3 months in obese subjects. In addition, the study will also determine early efficacy signals for various metabolic diseases associated with elevated triglycerides and / or obesity, and whether V103 shows clinical safety and efficacy characteristics as a therapeutic agent suitable for the development of metabolic diseases of hypertriglyceridemia, obesity / or non-alcoholic fatty liver disease.

[0191] Goals and End Points

[0192] The primary objective or objectives are to evaluate the safety and tolerability of V103 in obese subjects following repeated dosing via subcutaneous (SC) injection over 12 weeks. Endpoints related to this objective include:

[0193] Adverse events (AEs)

[0194] Vital signs

[0195] Liver function tests: aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin (total bile), and alkaline phosphatase (ALP)

[0196] 24-hour urine cortisol

[0197] Electrolytes

[0198] Secondary objectives include:

[0199] To evaluate the effect of V103 on triglycerides and lipid profiles at 12 weeks of treatment.

[0200] To evaluate the potential effects of V103 on bone biomarkers of resorption, bone formation and the balance of resorption and deposition at 12 weeks of treatment.

[0201] • To evaluate the effect of V103 on body weight as measured by body weight, body mass index (BMI), and waist circumference at 12 weeks.

[0202] End points related to these goals include:

[0203] Total cholesterol, LDL-C, HDL-C, and triglycerides (fasting) at 12 weeks

[0204] Biomarkers of bone resorption at 12 weeks (serum CTX-1, urine NTX-1)

[0205] and biomarkers of bone formation (serum BSAP, P1NP, and osteocalcin) at 12 weeks

[0206] Body weight, body mass index (BMI), waist circumference, and liver fat percentage at 12 weeks as measured by MRI

[0207] Other research objectives include:

[0208] • Evaluation of the pharmacokinetics (PK) of V103 in obese subjects after 12 weeks of repeated dosing.

[0209] • Evaluation of the immunogenicity of V103 following repeated SC administration of V103.

[0210] To evaluate the potential glucose metabolism effect of V103.

[0211] Non-limiting examples of endpoints related to these other study objectives include:

[0212] PK parameters will be determined, including: AUC 最后 , C 最大 and T 最大 ;

[0213] Fasting glucose, insulin, glucagon, and C-peptide, as well as extrapolated estimates of insulin sensitivity and secretion (HOMA) for up to 12 weeks;

[0214] HbA1c, glycated albumin;

[0215] Adiponectin; and

[0216] Pre-dose and post-dose anti-drug antibodies (ADA) through 26 weeks.

[0217] Study Design :

[0218] The study was designed as a non-confirmatory, multicenter, randomized, investigator- and subject-blinded, placebo-controlled safety study in obese subjects, in which V103 or placebo was administered subcutaneously with repeated dosing over 3 months.

[0219] Approximately 60 subjects will be randomized (1:1, active:placebo) to receive V103 300 mg (once every 4 weeks for three doses) or matching placebo by subcutaneous (SC) injection.

[0220] The trial consists of three main periods: (1) screening, lasting 1 to 4 weeks, (2) a subject- and investigator-blinded randomized treatment period, expected to last 12 weeks, and (3) end-of-study assessments, approximately 120 days (approximately 6 half-lives) after the last dose of study drug.

[0221] Group Expansion: Following an interim analysis (e.g., after approximately 15 subjects in each group reach end of treatment (Day 84)), if there is evidence of efficacy and an additional dose needs to be evaluated, lower dose groups may be evaluated to inform future studies prior to population expansion.

[0222] group :

[0223] The study population will include approximately 60 adult male and female obese subjects between 18 and 55 years of age, inclusive.

[0224] Key inclusion criteria:

[0225] Male and female subjects aged 18 to 55 years (inclusive);

[0226] Body mass index (BMI) between 30 and 45 kg / m 2 BMI = weight (kg) / [height (m)] 2 , inclusive; and

[0227] Screening triglycerides 150-500 mg / dL (1.69-5.65 mmol / L), inclusive.

[0228] Key exclusion criteria:

[0229] History of hepatobiliary disease, cholelithiasis, or cholestasis (on medical history or ultrasound screening), hepatic encephalopathy, esophageal varices, or portal vein shunts;

[0230] Liver disease or liver damage as indicated by liver function tests (ALT, AST, GGT, alkaline phosphatase, or serum bilirubin) above the upper limit of normal at screening or baseline;

[0231] Chronic infection with human immunodeficiency virus (HIV), hepatitis B (HBV), or hepatitis C (HCV). Subjects are excluded by a positive HBV surface antigen (HBsAg) test, or a positive HBV core antigen test under standard local practice. Subjects with a positive HCV antibody test should have HCV RNA levels measured. Subjects with positive (detectable) HCV RNA should be excluded;

[0232] Fasting triglycerides greater than or equal to 500 mg / dL [5.65 mmol / L] or concurrent use of medications to treat hypertriglyceridemia (fibrates, omega-3 fatty acids, niacin);

[0233] History of pancreatic injury or pancreatitis or other pancreatic disease; amylase or lipase above ULN at screening or baseline;

[0234] History of hypersensitivity reaction to similar biologics, FGF21 protein analogs, or Fc fusion proteins;

[0235] · History of bone disorders (including but not limited to osteoporosis, osteopenia, osteomalacia, severe vitamin D deficiency);

[0236] Plasma 25-hydroxyvitamin D level below the lower limit of the normal range at screening;

[0237] Contraindications to MRI

[0238] Weight change (self-reported change of more than 5% or self-reported change of 5 kg in the past 3 months);

[0239] Use of weight-loss medications such as orlistat (Xenical, Alli), lorcaserin (Belviq), phentermine-topiramate (Qsymia), naltrexone-bupropion (Contrave), or liraglutide (Victoza) toza or laglutide (Saxenda) or other glucagon-like peptide-1 (GLP1) receptor agonists (exenatide (Byetta / Bydureon), lixisenatide (Luxumia), albiglutide (Tanzeum), or dulaglutide (Trulicity or others); and

[0240] Patients were also excluded from following a diet, weight loss, or exercise program with the specific intention of losing weight, or any clinical diagnosis of an eating disorder within 3 months prior to randomization.

[0241] Pharmacokinetic assessment: C from serum concentration-time data, if available 最大 , T 最大 , C 最后 , T 最后 , AUC 最后 , AUCtau, AUCinf, T1 / 2, Vz / F and CL / F.

[0242] Efficacy / PD Assessment:

[0243] Fasting triglycerides

[0244] ·weight

[0245] Key safety assessments

[0246] Adverse event and serious adverse event monitoring

[0247] Bone biomarkers

[0248] Liver function

[0249] Ultrasound

[0250] Pancreatic function

[0251] Pituitary-adrenal function

[0252] Other pituitary-endocrine safety monitoring

[0253] Physical examination and vital signs

[0254] Monitoring of routine laboratory markers in blood and urine

[0255] Electrocardiogram

[0256] Columbia-Suicidality Severity Rating Scale (C-SSRS)

[0257] Mechanistic biomarkers and biomarkers of inflammation

[0258] Other embodiments

[0259] In one aspect, provided herein is a method for treating, preventing or managing a metabolic disorder or a cardiovascular disorder, the method comprising administering a human FGF21 protein variant to a subject in need thereof at a dose ranging from 100 mg to 600 mg.

[0260] In another aspect, provided herein is a method for treating, preventing or managing hypercholesterolemia, mixed dyslipidemia or hypertriglyceridemia, the method comprising administering a human FGF21 protein variant to a subject in need thereof at a dose ranging from 100 mg to 600 mg.

[0261] In another aspect, provided herein is a method of reducing body weight, the method comprising administering a human FGF21 protein variant to a subject in need thereof at a dose ranging from 100 mg to 600 mg.

[0262] In another aspect, provided herein is a method for lowering elevated LDL-C, total C, triglycerides and / or Apo B, the method comprising administering a human FGF21 protein variant to a subject in need thereof at a dose ranging from 100 mg to 600 mg.

[0263] In another aspect, provided herein is a method of increasing HDL-C, the method comprising administering a human FGF21 protein variant to a subject in need thereof at a dose ranging from 100 mg to 600 mg.

[0264] In another aspect, provided herein is a method of lowering triglyceride levels, the method comprising administering a human FGF21 protein variant to a subject in need thereof at a dose ranging from 100 mg to 600 mg.

[0265] In another aspect, provided herein is a method of reducing cardiovascular risk, the method comprising administering a human FGF21 protein variant to a subject in need thereof at a dose ranging from 100 mg to 600 mg.

[0266] In one aspect, provided herein is a human FGF21 protein variant for use in a method for treating, preventing or managing a metabolic disorder or a cardiovascular disorder, the method comprising administering the human FGF21 protein variant to a subject in need thereof at a dose ranging from 100 mg to 600 mg.

[0267] In one aspect, provided herein is a human FGF21 protein variant for use in a method for treating, preventing or managing hypercholesterolemia, mixed dyslipidemia or hypertriglyceridemia, the method comprising administering the human FGF21 protein variant to a subject in need thereof at a dose ranging from 100 mg to 600 mg.

[0268] In one aspect, provided herein is a human FGF21 protein variant for use in a method for reducing body weight, the method comprising administering the human FGF21 protein variant to a subject in need thereof at a dose ranging from 100 mg to 600 mg.

[0269] In one aspect, provided herein is a human FGF21 protein variant for use in a method for lowering elevated LDL-C, total C, triglycerides and / or Apo B, the method comprising administering the human FGF21 protein variant to a subject in need thereof at a dose ranging from 100 mg to 600 mg.

[0270] In one aspect, provided herein is a human FGF21 protein variant for use in a method of increasing HDL-C, the method comprising administering the human FGF21 protein variant to a subject in need thereof at a dose ranging from 100 mg to 600 mg.

[0271] In one aspect, provided herein is a human FGF21 protein variant for use in a method for lowering triglyceride levels, the method comprising administering the human FGF21 protein variant to a subject in need thereof at a dose ranging from 100 mg to 600 mg.

[0272] In one aspect, provided herein is a human FGF21 protein variant for use in a method of reducing cardiovascular risk, the method comprising administering the human FGF21 protein variant to a subject in need thereof at a dose ranging from 100 mg to 600 mg.

[0273] In one aspect, provided herein is a use of a human FGF21 protein variant in the preparation of a medicament for use in a method for treating, preventing or managing a metabolic disorder or a cardiovascular disorder, the method comprising administering the human FGF21 protein variant to a subject in need thereof at a dose ranging from 100 mg to 600 mg.

[0274] In one aspect, provided herein is the use of a human FGF21 protein variant in the preparation of a medicament for use in a method for treating, preventing or managing hypercholesterolemia, mixed dyslipidemia or hypertriglyceridemia, the method comprising administering the human FGF21 protein variant to a subject in need thereof at a dose ranging from 100 mg to 600 mg.

[0275] In one aspect, provided herein is a use of a human FGF21 protein variant in the preparation of a medicament for use in a method for reducing body weight, the method comprising administering the human FGF21 protein variant to a subject in need thereof at a dose ranging from 100 mg to 600 mg.

[0276] In one aspect, provided herein is a use of a human FGF21 protein variant in the preparation of a medicament for use in a method for lowering elevated LDL-C, total C, triglycerides and / or Apo B, the method comprising administering the human FGF21 protein variant to a subject in need thereof at a dose ranging from 100 mg to 600 mg.

[0277] In one aspect, provided herein is a use of a human FGF21 protein variant in the preparation of a medicament for use in a method for increasing HDL-C, the method comprising administering the human FGF21 protein variant to a subject in need thereof at a dose ranging from 100 mg to 600 mg.

[0278] In one aspect, provided herein is a use of a human FGF21 protein variant in the preparation of a medicament for use in a method for lowering triglyceride levels, the method comprising administering the human FGF21 protein variant to a subject in need thereof at a dose ranging from 100 mg to 600 mg.

[0279] In one aspect, provided herein is a use of a human FGF21 protein variant in the preparation of a medicament for use in a method for reducing cardiovascular risk, the method comprising administering the human FGF21 protein variant to a subject in need thereof at a dose ranging from 100 mg to 600 mg.

[0280] In one aspect, provided herein is a use of a human FGF21 protein variant in the preparation of a medicament for use in a method for reducing cardiovascular risk, the method comprising administering the human FGF21 protein variant to a subject in need thereof at a dose ranging from 100 mg to 600 mg.

[0281] In one aspect, provided herein is the use of a human FGF21 protein variant in an amount ranging from 100 mg to 600 mg in the preparation of a medicament for use in a method of treating, preventing or managing a metabolic disorder or a cardiovascular disorder.

[0282] In one aspect, provided herein is the use of a human FGF21 protein variant in an amount ranging from 100 mg to 600 mg in the preparation of a medicament for use in a method of treating, preventing or managing hypercholesterolemia, mixed dyslipidemia, or hypertriglyceridemia.

[0283] In one aspect, provided herein is the use of a human FGF21 protein variant in an amount ranging from 100 mg to 600 mg in the preparation of a medicament for use in a method of reducing body weight.

[0284] In one aspect, provided herein is the use of a human FGF21 protein variant in an amount ranging from 100 mg to 600 mg in the preparation of a medicament for use in a method of lowering elevated LDL-C, total C, triglycerides and / or Apo B.

[0285] In one aspect, provided herein is the use of a human FGF21 protein variant in an amount ranging from 100 mg to 600 mg in the preparation of a medicament for use in a method of increasing HDL-C.

[0286] In one aspect, provided herein is the use of a human FGF21 protein variant in an amount ranging from 100 mg to 600 mg in the preparation of a medicament for use in a method of lowering triglyceride levels.

[0287] In one embodiment, the method reduces triglyceride levels by at least about 40% or at least about 50%.

[0288] In one aspect, provided herein is the use of a human FGF21 protein variant in an amount ranging from 100 mg to 600 mg in the preparation of a medicament for use in a method of reducing cardiovascular risk.

[0289] In one aspect, provided herein is the use of a human FGF21 protein variant in an amount ranging from 100 mg to 600 mg in the manufacture of a medicament for use in the treatment, prevention or management of a metabolic disorder or a cardiovascular disorder.

[0290] In one aspect, provided herein is the use of a human FGF21 protein variant in an amount ranging from 100 mg to 600 mg in the manufacture of a medicament for use in the treatment, prevention or management of hypercholesterolemia, mixed dyslipidemia, or hypertriglyceridemia.

[0291] In one aspect, provided herein is the use of a human FGF21 protein variant in an amount ranging from 100 mg to 600 mg in the manufacture of a medicament for use in weight loss.

[0292] In one aspect, provided herein is the use of a human FGF21 protein variant in an amount ranging from 100 mg to 600 mg in the manufacture of a medicament for use in lowering elevated LDL-C, total C, triglycerides and / or Apo B.

[0293] In one aspect, provided herein is the use of a human FGF21 protein variant in an amount ranging from 100 mg to 600 mg in the manufacture of a medicament for use in increasing HDL-C.

[0294] In one aspect, provided herein is the use of a human FGF21 protein variant in an amount ranging from 100 mg to 600 mg in the manufacture of a medicament for use in lowering triglycerides.

[0295] In one embodiment, the method reduces triglyceride levels by at least about 40% or at least about 50%.

[0296] In one aspect, provided herein is the use of a human FGF21 protein variant in an amount ranging from 100 mg to 600 mg in the manufacture of a medicament for use in reducing cardiovascular risk.

[0297] In one aspect, provided herein is a use of a human FGF21 protein variant in the manufacture of a medicament for treating a metabolic disorder or a cardiovascular disorder, wherein the unit dose of the human FGF21 protein variant is in the range of 100 mg to 600 mg.

[0298] In one aspect, provided herein is a use of a human FGF21 protein variant in the manufacture of a medicament for treating hypercholesterolemia, mixed dyslipidemia or hypertriglyceridemia, wherein the unit dose of the human FGF21 protein variant is in the range of 100 mg to 600 mg.

[0299] In one aspect, provided herein is a use of a human FGF21 protein variant in the manufacture of a medicament for weight loss, wherein the unit dose of the human FGF21 protein variant is in the range of 100 mg to 600 mg.

[0300] In one aspect, provided herein is the use of a human FGF21 protein variant in the manufacture of a medicament for lowering LDL-C, total C, triglycerides and / or Apo B levels, wherein the unit dose of the human FGF21 protein variant is in the range of 100 mg to 600 mg.

[0301] In one aspect, provided herein is a use of a human FGF21 protein variant in the manufacture of a medicament for use in a method for increasing HDL-C, wherein the unit dose of the human FGF21 protein variant is in the range of 100 mg to 600 mg.

[0302] In one aspect, provided herein is a use of a human FGF21 protein variant in the manufacture of a medicament for lowering triglyceride levels, wherein the unit dose of the human FGF21 protein variant is in the range of 100 mg to 600 mg.

[0303] In one aspect, provided herein is a use of a human FGF21 protein variant in the manufacture of a medicament for use in a method for reducing cardiovascular risk, wherein the unit dose of the human FGF21 protein variant is in the range of 100 mg to 600 mg.

[0304] In one aspect, provided herein is a human FGF21 protein variant for use in a method for treating, preventing or managing a metabolic disorder or cardiovascular disorder of a human subject, wherein the human FGF21 protein variant is provided for administration with a dosage in the range of 100 mg to 600 mg. In one aspect, provided herein is a human FGF21 protein variant for use in a method for treating, preventing or managing a metabolic disorder or cardiovascular disorder of a human subject, wherein the human FGF21 protein variant is provided for administration with an amount in the range of 100 mg to 600 mg.

[0305] In some embodiments of these aspects, the metabolic disorder or cardiovascular disorder is selected from hypercholesterolemia, dyslipidemia, hypertriglyceridemia, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), type 2 diabetes, and obesity.

[0306] In some embodiments of these aspects, treating, preventing, or managing a metabolic or cardiovascular disorder comprises or is characterized by reducing one or more of: a subject's body weight, liver fat content, elevated LDL-C, total C, triglycerides, and Apo B levels. In certain embodiments, treating, preventing, or managing a metabolic or cardiovascular disorder comprises or is characterized by increasing a subject's HDL-C levels.

[0307] In some embodiments of these aspects, treating, preventing, or managing a metabolic disorder or a cardiovascular disorder comprises or is characterized by reducing a subject's triglyceride levels by at least about 40% or by at least about 50%. In some embodiments, treating, preventing, or managing a metabolic disorder or a cardiovascular disorder comprises or is characterized by reducing a subject's triglyceride levels by at least about 40%. In some embodiments, treating, preventing, or managing a metabolic disorder or a cardiovascular disorder comprises or is characterized by reducing a subject's triglyceride levels by at least about 50%.

[0308] In some embodiments of these aspects, treating, preventing, or managing a metabolic disorder or a cardiovascular disorder comprises or is characterized by reducing cardiovascular risk in the subject.

[0309] In some embodiments of these aspects, the metabolic disorder or cardiovascular disorder is dyslipidemia (optionally mixed dyslipidemia), hypertriglyceridemia (optionally severe hypertriglyceridemia), or hypercholesterolemia (optionally primary hypercholesterolemia). In some embodiments, the metabolic disorder or cardiovascular disorder is non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH). In certain embodiments, the subject is 18 to 55 years old. In some embodiments, the subject has a body mass index (BMI) of 30 to 45 kg / m 2 In some embodiments, the subject has a body mass index (BMI) of 30 to 45 kg / m 2 In some embodiments, the subject has a triglyceride level within the range of 150-500 mg / dL (1.69-5.65 mmol / L) when measured before administration of the human FGF21 protein variant.

[0310] In some embodiments of these aspects, the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G and G202A numbered according to SEQ ID NO: 1. In certain embodiments, the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11.

[0311] In some embodiments of these aspects, human FGF21 protein variants are provided for use at a dosage of at least 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, or 400 mg. In certain embodiments, human FGF21 protein variants are provided for use at a dosage of about 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, or 190 mg, optionally wherein the human FGF21 protein variants are provided at a dosage of about 100 mg or 150 mg. In some embodiments, human FGF21 protein variants are provided for administration at a dosage of about 200mg, 210mg, 220mg, 230mg, 240mg, 250mg, 260mg, 270mg, 280mg, 290mg, 300mg, 310mg, 320mg, 330mg, 340mg, or 350mg, optionally wherein the human FGF21 protein variants are provided at a dosage of about 200mg, 250mg, or 300mg. In some embodiments, human FGF21 protein variants are provided for administration at a dosage of at least 100mg. In some embodiments, human FGF21 protein variants are provided for administration at a dosage of about 100mg. In some embodiments, human FGF21 protein variants are provided for administration at a dosage of at least 150mg. In some embodiments, human FGF21 protein variants are provided for administration at a dosage of about 150mg. In some embodiments, human FGF21 protein variants are provided for administration at a dosage of at least 200mg. In some embodiments, human FGF21 protein variants are provided for administration at a dosage of about 200 mg. In some embodiments, human FGF21 protein variants are provided for administration at a dosage of at least 250 mg. In some embodiments, human FGF21 protein variants are provided for administration at a dosage of about 250 mg. In some embodiments, human FGF21 protein variants are provided for administration at a dosage of 250 mg. In some embodiments, human FGF21 protein variants are provided for administration at a dosage of at least 300 mg. In some embodiments, human FGF21 protein variants are provided for administration at a dosage of about 300 mg. In some embodiments, human FGF21 protein variants are provided for administration at a dosage of 300 mg.

[0312] In some embodiments of these aspects, human FGF21 protein variants are provided for use in an amount of at least 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, or 400 mg. In certain embodiments, human FGF21 protein variants are provided for use in an amount of about 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, or 190 mg, optionally wherein the human FGF21 protein variants are provided in an amount of about 100 mg or 150 mg. In some embodiments, human FGF21 protein variants are provided for administration in an amount of about 200mg, 210mg, 220mg, 230mg, 240mg, 250mg, 260mg, 270mg, 280mg, 290mg, 300mg, 310mg, 320mg, 330mg, 340mg, or 350mg, optionally wherein the human FGF21 protein variants are provided in an amount of about 200mg, 250mg, or 300mg. In some embodiments, human FGF21 protein variants are provided for administration in an amount of at least 100mg. In some embodiments, human FGF21 protein variants are provided for administration in an amount of about 100mg. In some embodiments, human FGF21 protein variants are provided for administration in an amount of at least 150mg. In some embodiments, human FGF21 protein variants are provided for administration in an amount of about 150mg. In some embodiments, human FGF21 protein variants are provided for administration in an amount of at least 200mg. In some embodiments, human FGF21 protein variants are provided for administration in an amount of about 200 mg. In some embodiments, human FGF21 protein variants are provided for administration in an amount of at least 250 mg. In some embodiments, human FGF21 protein variants are provided for administration in an amount of about 250 mg. In some embodiments, human FGF21 protein variants are provided for administration in an amount of 250 mg. In some embodiments, human FGF21 protein variants are provided for administration in an amount of at least 300 mg. In some embodiments, human FGF21 protein variants are provided for administration in an amount of about 300 mg. In some embodiments, human FGF21 protein variants are provided for administration in an amount of 300 mg.

[0313] In some embodiments, human FGF21 protein variants are provided in the form of subcutaneous administration. In some embodiments, human FGF21 protein variants are provided for administration once a month or once every 4 weeks, once every 3 weeks, once every 2 weeks, or once a week. In some embodiments, human FGF21 protein variants are provided for administration once a month or once every 4 weeks, once every 3 weeks, once every 2 weeks, or once a week. In some embodiments, human FGF21 protein variants are provided in the form of subcutaneous administration.

[0314] In one aspect, provided herein is a method for treating, preventing or managing a metabolic disorder or cardiovascular disorder in a human subject, wherein the method includes administering human FGF21 protein variants to a subject at a dosage in the range of 100 mg to 600 mg. In one aspect, provided herein is a method for treating, preventing or managing a metabolic disorder or cardiovascular disorder in a human subject, wherein the method includes administering human FGF21 protein variants to a subject at an amount in the range of 100 mg to 600 mg.

[0315] In some embodiments, the metabolic disorder or cardiovascular disorder is selected from hypercholesterolemia, dyslipidemia, hypertriglyceridemia, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), type 2 diabetes, and obesity.

[0316] In some embodiments, the treatment, prevention, or management of a metabolic or cardiovascular disorder comprises or is characterized by reducing one or more of the following: a subject's body weight, liver fat content, elevated LDL-C, total C, triglycerides, and Apo B levels. In certain embodiments, the treatment, prevention, or management of a metabolic or cardiovascular disorder comprises or is characterized by increasing a subject's HDL-C levels.

[0317] In some embodiments, the treatment, prevention, or management of a metabolic disorder or a cardiovascular disorder comprises or is characterized by reducing the triglyceride level of a subject by at least about 40% or at least about 50%. In some embodiments, the treatment, prevention, or management of a metabolic disorder or a cardiovascular disorder comprises or is characterized by reducing the triglyceride level of a subject by at least about 40%. In some embodiments, the treatment, prevention, or management of a metabolic disorder or a cardiovascular disorder comprises or is characterized by reducing the triglyceride level of a subject by at least about 50%.

[0318] In some embodiments, treating, preventing, or managing a metabolic disorder or a cardiovascular disorder comprises or is characterized by reducing cardiovascular risk in the subject.

[0319] In some embodiments, the metabolic disorder or cardiovascular disorder is dyslipidemia, hypertriglyceridemia or hypercholesterolemia. In some embodiments, the metabolic disorder or cardiovascular disorder is dyslipidemia. In some embodiments, the metabolic disorder or cardiovascular disorder is hypertriglyceridemia. In some embodiments, the metabolic disorder or cardiovascular disorder is hypercholesterolemia. In some embodiments, the metabolic disorder or cardiovascular disorder is mixed dyslipidemia. In some embodiments, the metabolic disorder or cardiovascular disorder is severe hypertriglyceridemia. In some embodiments, the metabolic disorder or cardiovascular disorder is primary hypercholesterolemia. In some embodiments, the metabolic disorder or cardiovascular disorder is non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH). In some embodiments, the metabolic disorder or cardiovascular disorder is non-alcoholic fatty liver disease (NAFLD). In some embodiments, the metabolic disorder or cardiovascular disorder is non-alcoholic steatohepatitis (NASH). In certain embodiments, the subject is 18 to 55 years old. In some embodiments, the subject's body mass index (BMI) is between 30 and 45 kg / m 2 In some embodiments, the subject has a body mass index (BMI) of ≥ 27.5.

[0320] In one aspect, provided herein is a human FGF21 protein variant for use in a method for treating, preventing or managing hypercholesterolemia in a human subject, wherein the human FGF21 protein variant is administered at 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, 350 mg, 360 mg, 370 mg, 380 mg, 390 mg, 400 mg, 410 mg, 420 mg, 430 mg, 440 mg, 450 mg, 460 mg, 470 mg, 480 mg, 490 mg, 500 mg, 510 mg, 520 mg, 530 mg, 540 mg, 550 mg, 560 mg, 570 mg, 580 mg, 590 mg, 600 mg, 610 mg, 620 mg, 630 mg, 640 mg, 650 mg, 660 mg, 670 mg, 680 mg, 690 mg, 700 mg, 710 mg, 720 mg, 730 mg, 740 mg 00mg, 310mg, 320mg, 330mg, 340mg, or 350mg for administration, optionally wherein the human FGF21 protein variant is provided at a dose of about 100mg, 150mg, 200mg, 250mg, or 300mg, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1. Optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11. In one embodiment thereof, human FGF21 variants are provided for administration at a dosage of about 100mg, 150mg, 200mg, 250mg or 300mg once a week. In one embodiment thereof, human FGF21 variants are provided for administration at a dosage of about 100mg, 150mg, 200mg, 250mg or 300mg once every two weeks. In one embodiment thereof, human FGF21 variants are provided for administration once every three weeks at a dosage of about 100mg, 150mg, 200mg, 250mg or 300mg. In one embodiment thereof, human FGF21 variants are provided for administration once every four weeks or once a month at a dosage of about 100mg, 150mg, 200mg, 250mg or 300mg.

[0321] In one aspect, provided herein is a human FGF21 protein variant for use in a method for treating, preventing or managing dyslipidemia in a human subject, wherein the human FGF21 protein variant is administered at 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, 350 mg, 360 mg, 370 mg, 380 mg, 390 mg, 400 mg, 410 mg, 420 mg, 430 mg, 440 mg, 450 mg, 460 mg, 470 mg, 480 mg, 490 mg, 500 mg, 510 mg, 520 mg, 530 mg, 540 mg, 550 mg, 560 mg, 570 mg, 580 mg, 590 mg, 600 mg, 610 mg, 620 mg, 630 mg, 640 mg, 650 mg, 660 mg, 670 mg, 680 mg, 690 mg, 700 mg, 710 mg, 720 mg, 730 mg, 740 mg 00mg, 310mg, 320mg, 330mg, 340mg, or 350mg for administration, optionally wherein the human FGF21 protein variant is provided at a dose of about 100mg, 150mg, 200mg, 250mg, or 300mg, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11. In one embodiment thereof, human FGF21 variants are provided for administration at a dosage of about 100mg, 150mg, 200mg, 250mg or 300mg once a week. In one embodiment thereof, human FGF21 variants are provided for administration at a dosage of about 100mg, 150mg, 200mg, 250mg or 300mg once every two weeks. In one embodiment thereof, human FGF21 variants are provided for administration once every three weeks at a dosage of about 100mg, 150mg, 200mg, 250mg or 300mg. In one embodiment thereof, human FGF21 variants are provided for administration once every four weeks or once a month at a dosage of about 100mg, 150mg, 200mg, 250mg or 300mg.

[0322] In one aspect, provided herein is a human FGF21 protein variant for use in a method for treating, preventing or managing hypertriglyceridemia in a human subject, wherein the human FGF21 protein variant is administered at 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, 350 mg, 360 mg, 370 mg, 380 mg, 390 mg, 400 mg, 410 mg, 420 mg, 430 mg, 440 mg, 450 mg, 460 mg, 470 mg, 480 mg, 490 mg, 500 mg, 510 mg, 520 mg, 530 mg, 540 mg, 550 mg, 560 mg, 570 mg, 580 mg, 590 mg, 600 mg, 610 mg, 620 mg, 630 mg, 640 mg, 650 mg, 660 mg, 670 mg, 680 mg, 690 mg, 700 mg, 710 mg, 720 mg, 730 mg, 740 mg 00mg, 310mg, 320mg, 330mg, 340mg, or 350mg for administration, optionally wherein the human FGF21 protein variant is provided at a dose of about 100mg, 150mg, 200mg, 250mg, or 300mg, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1. Optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11. In one embodiment thereof, human FGF21 variants are provided for administration at a dosage of about 100mg, 150mg, 200mg, 250mg or 300mg once a week. In one embodiment thereof, human FGF21 variants are provided for administration at a dosage of about 100mg, 150mg, 200mg, 250mg or 300mg once every two weeks. In one embodiment thereof, human FGF21 variants are provided for administration once every three weeks at a dosage of about 100mg, 150mg, 200mg, 250mg or 300mg. In one embodiment thereof, human FGF21 variants are provided for administration once every four weeks or once a month at a dosage of about 100mg, 150mg, 200mg, 250mg or 300mg.

[0323] In one aspect, provided herein is a human FGF21 protein variant for use in a method for treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) in a human subject, wherein the human FGF21 protein variant is administered at 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, 350 mg, 360 mg, 370 mg, 380 mg, 390 mg, 400 mg, 410 mg, 420 mg, 430 mg, 440 mg, 450 mg, 460 mg, 470 mg, 480 mg, 490 mg, 500 mg, 510 mg, 520 mg, 530 mg, 540 mg, 550 mg, 560 mg, 570 mg, 580 mg, 590 mg, 600 mg, 610 mg, 620 mg, 630 mg, 640 mg, 650 mg, 660 mg, 670 mg, 680 mg, 690 mg, 700 mg, 710 mg, 720 mg, 730 mg, 740 mg, 750 mg, 760 mg, 770 mg, 780 mg, 790 mg, 800 mg, 810 mg, A dose or amount within the range of 90 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg is provided for administration, optionally wherein the human FGF21 protein variant is provided at a dose of approximately 100 mg, 150 mg, 200 mg, 250 mg, or 300 mg, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11. In one embodiment thereof, human FGF21 variants are provided for administration at a dosage of about 100mg, 150mg, 200mg, 250mg or 300mg once a week. In one embodiment thereof, human FGF21 variants are provided for administration at a dosage of about 100mg, 150mg, 200mg, 250mg or 300mg once every two weeks. In one embodiment thereof, human FGF21 variants are provided for administration once every three weeks at a dosage of about 100mg, 150mg, 200mg, 250mg or 300mg. In one embodiment thereof, human FGF21 variants are provided for administration once every four weeks or once a month at a dosage of about 100mg, 150mg, 200mg, 250mg or 300mg.

[0324] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic steatohepatitis (NASH) in a human subject, wherein the human FGF21 protein variant is administered at 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, 350 mg, 360 mg, 370 mg, 380 mg, 390 mg, 400 mg, 410 mg, 420 mg, 430 mg, 440 mg, 450 mg, 460 mg, 470 mg, 480 mg, 490 mg, 500 mg, 510 mg, 520 mg, 530 mg, 540 mg, 550 mg, 560 mg, 570 mg, 580 mg, 590 mg, 600 mg, 610 mg, 620 mg, 630 mg, 640 mg, 650 mg, 660 mg, 670 mg, 680 mg, 690 mg, 700 mg, 710 mg, 720 mg, 730 mg, 740 mg In one embodiment, the human FGF21 variant is provided for administration at a dose or amount within the range of about 100 mg, 150 mg, 200 mg, 250 mg, or 300 mg, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, wherein the mature human FGF21 protein or a fragment thereof comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, and wherein the human FGF21 variant comprises the amino acid sequence of SEQ ID NO: 11. In one embodiment thereof, the human FGF21 variant is provided for administration once a week at a dose of about 100 mg, 150 mg, 200 mg, 250 mg, or 300 mg. In one embodiment thereof, human FGF21 variants are provided for administration at a dosage of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg once every two weeks. In one embodiment thereof, human FGF21 variants are provided for administration at a dosage of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg once every three weeks. In one embodiment thereof, human FGF21 variants are provided for administration at a dosage of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg once every four weeks or once a month.

[0325] In one aspect, provided herein is a human FGF21 protein variant for use in a method for treating, preventing or managing type 2 diabetes in a human subject, wherein the human FGF21 protein variant is administered at 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, 350 mg, 360 mg, 370 mg, 380 mg, 390 mg, 400 mg, 410 mg, 420 mg, 430 mg, 440 mg, 450 mg, 460 mg, 470 mg, 480 mg, 490 mg, 500 mg, 510 mg, 520 mg, 530 mg, 540 mg, 550 mg, 560 mg, 570 mg, 580 mg, 590 mg, 600 mg, 610 mg, 620 mg, 630 mg, 640 mg, 650 mg, 660 mg, 670 mg, 680 mg, 690 mg, 700 mg, 710 mg, 720 mg, 730 mg, 740 mg 00mg, 310mg, 320mg, 330mg, 340mg, or 350mg for administration, optionally wherein the human FGF21 protein variant is provided at a dose of about 100mg, 150mg, 200mg, 250mg, or 300mg, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11. In one embodiment thereof, human FGF21 variants are provided for administration at a dosage of about 100mg, 150mg, 200mg, 250mg or 300mg once a week. In one embodiment thereof, human FGF21 variants are provided for administration at a dosage of about 100mg, 150mg, 200mg, 250mg or 300mg once every two weeks. In one embodiment thereof, human FGF21 variants are provided for administration once every three weeks at a dosage of about 100mg, 150mg, 200mg, 250mg or 300mg. In one embodiment thereof, human FGF21 variants are provided for administration once every four weeks or once a month at a dosage of about 100mg, 150mg, 200mg, 250mg or 300mg.

[0326] In one aspect, provided herein is a human FGF21 protein variant for use in a method for treating, preventing or managing obesity in a human subject, wherein the human FGF21 protein variant is administered at 100 mg, 110 , 120mg, 130mg, 140mg, 150mg, 160mg, 170mg, 180mg, 190mg, 200mg, 210mg, 220mg, 230mg, 240mg, 250mg, 260mg, 270mg, 280mg, 290mg, 300mg, 310mg, 320mg, 330mg, 340mg, or 350mg for administration, optionally wherein the human FGF21 protein variant is provided at a dose of about 100mg, 150mg, 200mg, 250mg, or 300mg, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of: NO: 1 numbered Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11. In one embodiment thereof, the human FGF21 variant is provided for administration at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg once a week. In one embodiment thereof, the human FGF21 variant is provided for administration at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg once every two weeks. In one embodiment thereof, the human FGF21 variant is provided for administration at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg once every three weeks. In one embodiment thereof, the human FGF21 variant is provided for administration at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg once every four weeks or once a month.

[0327] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing hypercholesterolemia, dyslipidemia, hypertriglyceridemia, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), type 2 diabetes or obesity in a human subject, wherein the human FGF21 protein variant is provided for administration at a dose or amount in the range of 100 mg, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11. In one embodiment thereof, the human FGF21 variant is provided for once-weekly administration. In one embodiment thereof, human FGF21 variants are provided for administration once every two weeks. In one embodiment thereof, human FGF21 variants are provided for administration once every three weeks. In one embodiment thereof, human FGF21 variants are provided for administration once every four weeks or once a month.

[0328] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing hypercholesterolemia, dyslipidemia, hypertriglyceridemia, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), type 2 diabetes or obesity in a human subject, wherein the human FGF21 protein variant is provided for administration at a dose or amount within the range of 150 mg, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11. In one embodiment thereof, the human FGF21 variant is provided for once-weekly administration. In one embodiment thereof, human FGF21 variants are provided for administration once every two weeks. In one embodiment thereof, human FGF21 variants are provided for administration once every three weeks. In one embodiment thereof, human FGF21 variants are provided for administration once every four weeks or once a month.

[0329] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing hypercholesterolemia, dyslipidemia, hypertriglyceridemia, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), type 2 diabetes or obesity in a human subject, wherein the human FGF21 protein variant is provided for administration at a dose or amount within the range of 200 mg, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11. In one embodiment thereof, the human FGF21 variant is provided for once-weekly administration. In one embodiment thereof, human FGF21 variants are provided for administration once every two weeks. In one embodiment thereof, human FGF21 variants are provided for administration once every three weeks. In one embodiment thereof, human FGF21 variants are provided for administration once every four weeks or once a month.

[0330] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing hypercholesterolemia, dyslipidemia, hypertriglyceridemia, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), type 2 diabetes or obesity in a human subject, wherein the human FGF21 protein variant is provided for administration at a dose or amount within the range of 250 mg, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11. In one embodiment thereof, the human FGF21 variant is provided for once-weekly administration. In one embodiment thereof, human FGF21 variants are provided for administration once every two weeks. In one embodiment thereof, human FGF21 variants are provided for administration once every three weeks. In one embodiment thereof, human FGF21 variants are provided for administration once every four weeks or once a month.

[0331] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic steatohepatitis (NASH) in a human subject, wherein the human FGF21 protein variant is provided for administration at a dose or amount within the range of 300 mg, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises an amino acid sequence of SEQ ID NO: 11. In one embodiment thereof, the human FGF21 variant is provided for administration once a week. In one embodiment thereof, the human FGF21 variant is provided for administration once every two weeks. In one embodiment thereof, the human FGF21 variant is provided for administration once every three weeks. In one embodiment thereof, the human FGF21 variant is provided for administration once every four weeks or once a month.

[0332] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing hypercholesterolemia, dyslipidemia, hypertriglyceridemia, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), type 2 diabetes or obesity in a human subject, wherein the human FGF21 protein variant is provided for administration once a week at a dose or amount in the range of 100 mg or 150 mg, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, wherein the mature human FGF21 protein or a fragment thereof comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11.

[0333] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing hypercholesterolemia, dyslipidemia, hypertriglyceridemia, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), type 2 diabetes or obesity in a human subject, wherein the human FGF21 protein variant is provided for administration at a dose or amount in the range of 150 mg or 200 mg once every two weeks, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, wherein the mature human FGF21 protein or a fragment thereof comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11.

[0334] In one aspect, provided herein is a human FGF21 protein variant for use in a method for treating, preventing or managing hypercholesterolemia, dyslipidemia, hypertriglyceridemia, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), type 2 diabetes or obesity in a human subject, wherein the human FGF21 protein variant is provided for administration at a dose or amount within the range of 200 mg or 250 mg once every three weeks, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, wherein the mature human FGF21 protein or a fragment thereof comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11.

[0335] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing hypercholesterolemia, dyslipidemia, hypertriglyceridemia, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), type 2 diabetes or obesity in a human subject, wherein the human FGF21 protein variant is provided for administration at a dose or amount in the range of 250 mg or 300 mg once every four weeks or once monthly, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, wherein the mature human FGF21 protein or a fragment thereof comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11.

[0336] In one aspect, provided herein is a human FGF21 protein variant for use in a method for treating, preventing or managing hypercholesterolemia in a human subject, wherein the human FGF21 protein variant is provided for administration in combination with one or more additional therapeutically active agents, wherein the one or more additional therapeutically active agents are SGLT inhibitors selected from the group consisting of dapagliflozin, empagliflozin, canagliflozin, erpagliflozin, sogliaflozin, togliflozin, repagliflozin, lupagliflozin, ipagliaflozin, apagliflozin, bepagliflozin, hengagliflozin, licogliflozin and pharmaceutically acceptable salts of any of these, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of: according to SEQ ID Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A as numbered by SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11. In one embodiment thereof, the SGLT inhibitor is licogliflozin.

[0337] In one aspect, provided herein is a human FGF21 protein variant for use in a method for treating, preventing or managing dyslipidemia in a human subject, wherein the human FGF21 protein variant is provided for administration in combination with one or more additional therapeutically active agents, wherein the one or more additional therapeutically active agents are SGLT inhibitors selected from the group consisting of dapagliflozin, empagliflozin, canagliflozin, erpagliflozin, sogliaflozin, togliflozin, repagliflozin, lupagliflozin, ipagliaflozin, apagliflozin, bepagliflozin, hengagliflozin, licogliflozin and pharmaceutically acceptable salts of any of these, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of: according to SEQ ID Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A as numbered by SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11. In one embodiment thereof, the SGLT inhibitor is licogliflozin.

[0338] In one aspect, provided herein is a human FGF21 protein variant for use in a method for treating, preventing or managing hypertriglyceridemia in a human subject, wherein the human FGF21 protein variant is provided for administration in combination with one or more additional therapeutically active agents, wherein the one or more additional therapeutically active agents are SGLT inhibitors selected from the group consisting of dapagliflozin, empagliflozin, canagliflozin, erpagliflozin, sogliaflozin, togliflozin, repagliflozin, lupagliflozin, ipagliaflozin, apagliflozin, bepagliflozin, hengagliflozin, licogliflozin and pharmaceutically acceptable salts of any of these, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of: according to SEQ ID Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A as numbered by SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11. In one embodiment thereof, the SGLT inhibitor is licogliflozin.

[0339] In one aspect, provided herein is a human FGF21 protein variant for use in a method for treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) in a human subject, wherein the human FGF21 protein variant is provided for administration in combination with one or more additional therapeutically active agents, wherein the one or more additional therapeutically active agents are SGLT inhibitors selected from the group consisting of dapagliflozin, empagliflozin, canagliflozin, erpagliflozin, sogliaflozin, togliflozin, repagliflozin, lupagliflozin, ipagliaflozin, apagliflozin, bepagliflozin, hengagliflozin, licogliflozin and pharmaceutically acceptable salts of any of these, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of: according to SEQ ID Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A as numbered by SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11. In one embodiment thereof, the SGLT inhibitor is licogliflozin.

[0340] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic steatohepatitis (NASH) in a human subject, wherein the human FGF21 protein variant is provided for administration in combination with one or more additional therapeutically active agents, wherein the one or more additional therapeutically active agents are SGLT inhibitors selected from the group consisting of dapagliflozin, empagliflozin, canagliflozin, erpagliflozin, sogliaflozin, togliflozin, repagliflozin, lupagliflozin, ipagliaflozin, apagliflozin, bepagliflozin, hengagliflozin, licogliflozin and pharmaceutically acceptable salts of any of these, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of: according to SEQ ID Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A as numbered by SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11. In one embodiment thereof, the SGLT inhibitor is licogliflozin.

[0341] In one aspect, provided herein is a human FGF21 protein variant for use in a method for treating, preventing or managing type 2 diabetes in a human subject, wherein the human FGF21 protein variant is provided for administration in combination with one or more additional therapeutically active agents, wherein the one or more additional therapeutically active agents are SGLT inhibitors selected from the group consisting of dapagliflozin, empagliflozin, canagliflozin, erpagliflozin, sogliaflozin, togliflozin, repagliflozin, lupagliflozin, ipagliaflozin, apagliflozin, bepagliflozin, hengagliflozin, licogliflozin and pharmaceutically acceptable salts of any of these, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of: according to SEQ ID Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A as numbered by SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11. In one embodiment thereof, the SGLT inhibitor is licogliflozin.

[0342] In one aspect, provided herein is a human FGF21 protein variant for use in a method for treating, preventing or managing obesity in a human subject, wherein the human FGF21 protein variant is provided for administration in combination with one or more additional therapeutically active agents, wherein the one or more additional therapeutically active agents are SGLT inhibitors selected from the group consisting of dapagliflozin, empagliflozin, canagliflozin, erpagliflozin, sogliaflozin, togliflozin, repagliflozin, lupagliflozin, ipagliaflozin, apagliflozin, bepagliflozin, hengagliflozin, licogliflozin and pharmaceutically acceptable salts of any of these, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of: according to SEQ ID Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A as numbered by SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11. In one embodiment thereof, the SGLT inhibitor is licogliflozin.

[0343] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 protein variant is provided for administration once a week at a dose or amount within the range of 100 mg, and wherein the subject's liver fat content is reduced to within a manageable range or normal range as determined by a clinician or clinical guidelines or is reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises SEQ ID NO: Amino acid sequence of NO:11.

[0344] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 protein variant is provided for administration at a dose or amount within the range of 100 mg once every two weeks, and wherein the subject's liver fat content is reduced to within a manageable range or normal range as determined by a clinician or clinical guidelines or is reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises SEQ ID NO: Amino acid sequence of NO:11.

[0345] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 protein variant is provided for administration at a dose or amount within the range of 100 mg once every three weeks, and wherein the subject's liver fat content is reduced to within a manageable range or normal range as determined by a clinician or clinical guidelines or is reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises SEQ ID NO: Amino acid sequence of NO:11.

[0346] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 protein variant is provided for administration at a dose or amount within the range of 100 mg once every four weeks, and wherein the subject's liver fat content is reduced to within a manageable range or normal range as determined by a clinician or clinical guidelines or is reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises SEQ ID NO: Amino acid sequence of NO:11.

[0347] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 protein variant is provided for administration once a week at a dose or amount within the range of 150 mg, and wherein the subject's liver fat content is reduced to within a manageable range or normal range as determined by a clinician or clinical guidelines or is reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises SEQ ID NO: Amino acid sequence of NO:11.

[0348] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 protein variant is provided for administration at a dose or amount within the range of 150 mg once every two weeks, and wherein the subject's liver fat content is reduced to within a manageable range or normal range as determined by a clinician or clinical guidelines or is reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises SEQ ID NO: Amino acid sequence of NO:11.

[0349] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 protein variant is provided for administration at a dose or amount within the range of 150 mg once every three weeks, and wherein the subject's liver fat content is reduced to within a manageable range or normal range as determined by a clinician or clinical guidelines or is reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises SEQ ID NO: Amino acid sequence of NO:11.

[0350] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 protein variant is provided for administration at a dose or amount within the range of 150 mg once every four weeks, and wherein the subject's liver fat content is reduced to within a manageable range or normal range as determined by a clinician or clinical guidelines or is reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises SEQ ID NO: Amino acid sequence of NO:11.

[0351] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 protein variant is provided for administration once a week at a dose or amount within the range of 200 mg, and wherein the liver fat content of the subject is reduced to within a manageable range or normal range as determined by a clinician or clinical guidelines or is reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises SEQ ID NO: Amino acid sequence of NO:11.

[0352] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 protein variant is provided for administration at a dose or amount within the range of 200 mg once every two weeks, and wherein the subject's liver fat content is reduced to within a manageable range or normal range as determined by a clinician or clinical guidelines or is reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises SEQ ID NO: Amino acid sequence of NO:11.

[0353] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 protein variant is provided for administration at a dose or amount within the range of 200 mg once every three weeks, and wherein the subject's liver fat content is reduced to within a manageable range or normal range as determined by a clinician or clinical guidelines or is reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises SEQ ID NO: Amino acid sequence of NO:11.

[0354] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 protein variant is provided for administration at a dose or amount within the range of 200 mg once every four weeks, and wherein the subject's liver fat content is reduced to within a manageable range or normal range as determined by a clinician or clinical guidelines or is reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises SEQ ID NO: Amino acid sequence of NO:11.

[0355] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 protein variant is provided for administration once a week at a dose or amount within the range of 250 mg, and wherein the subject's liver fat content is reduced to within a manageable range or normal range as determined by a clinician or clinical guidelines or is reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises SEQ ID NO: Amino acid sequence of NO:11.

[0356] In one aspect, provided herein is a human FGF21 protein variant for use in a method for treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 protein variant is provided for administration at a dose or amount within the range of 250 mg once every two weeks, and wherein the subject's liver fat content is reduced to within a manageable range or normal range as determined by a clinician or clinical guidelines or is reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises SEQ ID NO: Amino acid sequence of NO:11.

[0357] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 protein variant is provided for administration at a dose or amount within the range of 250 mg once every three weeks, and wherein the subject's liver fat content is reduced to within a manageable range or normal range as determined by a clinician or clinical guidelines or is reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises SEQ ID NO: Amino acid sequence of NO:11.

[0358] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 protein variant is provided for administration at a dose or amount within the range of 250 mg once every four weeks, and wherein the subject's liver fat content is reduced to within a manageable range or normal range as determined by a clinician or clinical guidelines or is reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises SEQ ID NO: Amino acid sequence of NO:11.

[0359] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 protein variant is provided for administration once a week at a dose or amount within the range of 300 mg, and wherein the subject's liver fat content is reduced to within a manageable range or normal range as determined by a clinician or clinical guidelines or is reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises SEQ ID NO: Amino acid sequence of NO:11.

[0360] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 protein variant is provided for administration at a dose or amount within the range of 300 mg once every two weeks, and wherein the subject's liver fat content is reduced to within a manageable range or normal range as determined by a clinician or clinical guidelines or is reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises SEQ ID NO: Amino acid sequence of NO:11.

[0361] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 protein variant is provided for administration at a dose or amount within the range of 300 mg once every three weeks, and wherein the subject's liver fat content is reduced to within a manageable range or normal range as determined by a clinician or clinical guidelines or is reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises SEQ ID NO: Amino acid sequence of NO:11.

[0362] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 protein variant is provided for administration at a dose or amount within the range of 300 mg once every four weeks, and wherein the subject's liver fat content is reduced to within a manageable range or normal range as determined by a clinician or clinical guidelines or is reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises SEQ ID NO: Amino acid sequence of NO:11.

[0363] In one aspect, provided herein is a human FGF21 protein variant for use in a method for treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) in a human subject, wherein the human FGF21 protein variant is provided for administration in combination with licogliflozin or a pharmaceutically acceptable salt thereof, and wherein the subject's liver fat content is reduced to within a manageable range or normal range as determined by a clinician or clinical guidelines or is reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11.

[0364] In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic steatohepatitis (NASH) in a human subject, wherein the human FGF21 protein variant is provided for administration in combination with licogliflozin or a pharmaceutically acceptable salt thereof, and wherein the subject's liver fat content is reduced to within a manageable range or normal range as determined by a clinician or clinical guidelines or is reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11.

[0365] In one aspect, provided herein is a human FGF21 protein variant for use in a method for treating, preventing or managing triglyceridemia, wherein the human FGF21 protein variant is provided for administration at a dose or amount within the range of 100 mg once a week, once every two weeks, once every three weeks, once every four weeks, or once a month, and wherein the subject's triglyceride levels are reduced by at least about 40% or at least about 50%, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11.

[0366] In one aspect, provided herein is a human FGF21 protein variant for use in a method for treating, preventing or managing triglyceridemia, wherein the human FGF21 protein variant is provided for administration at a dose or amount within the range of 150 mg once a week, once every two weeks, once every three weeks, once every four weeks, or once a month, and wherein the subject's triglyceride levels are reduced by at least about 40% or at least about 50%, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11.

[0367] In one aspect, provided herein is a human FGF21 protein variant for use in a method for treating, preventing or managing triglyceridemia, wherein the human FGF21 protein variant is provided for administration at a dose or amount within the range of 200 mg once a week, once every two weeks, once every three weeks, once every four weeks, or once a month, and wherein the subject's triglyceride levels are reduced by at least about 40% or at least about 50%, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11.

[0368] In one aspect, provided herein is a human FGF21 protein variant for use in a method for treating, preventing or managing triglyceridemia, wherein the human FGF21 protein variant is provided for administration at a dose or amount within the range of 250 mg once a week, once every two weeks, once every three weeks, once every four weeks, or once a month, and wherein the subject's triglyceride levels are reduced by at least about 40% or at least about 50%, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11.

[0369] In one aspect, provided herein is a human FGF21 protein variant for use in a method for treating, preventing or managing triglyceridemia, wherein the human FGF21 protein variant is provided for administration at a dose or amount within the range of 300 mg once a week, once every two weeks, once every three weeks, once every four weeks, or once a month, and wherein the subject's triglyceride levels are reduced by at least about 40% or at least about 50%, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11.

[0370] Each of the foregoing aspects and embodiments, as well as other elements described herein, may be combined in any manner and without limitation.

Claims

1. Use of a human FGF21 protein variant for preparing a medicament for treating, preventing or managing metabolic disorders or cardiovascular disorders in a human subject, wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11 and is formulated for administration at a dose ranging from 100 mg to 600 mg, The pharmaceutical composition comprising the FGF21 protein variant is provided as a lyophilized formulation for reconstitution into a solution before administration, wherein the solution comprises 50 mg / mL to 150 mg / mL of human FGF21 protein variant, tromethamine buffer, sucrose and polysorbate 20, and The metabolic disorder or cardiovascular disorder is selected from hypercholesterolemia, dyslipidemia, hypertriglyceridemia, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), type 2 diabetes and obesity.

2. The use according to claim 1, wherein the metabolic disorder or cardiovascular disorder is non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH).

3. The method according to claim 1, wherein the subject has a body mass index (BMI) of 30 to 45 kg / m 2 The inclusive range was ≥27.5 for subjects of Asian origin or ancestry, adjusted for race.

4. The use according to claim 1, wherein the subject has a triglyceride level in the range of 150-500 mg / dL (1.69-5.65 mmol / L) when measured before administration of the human FGF21 protein variant.

5. The use according to any one of claims 1 to 4, wherein the human FGF21 protein variant is provided in combination with one or more additional therapeutically active agents.

6. The use according to claim 5, wherein the one or more additional therapeutically active agents are selected from the group consisting of compounds used in obesity therapy, diuretics, beta-blockers, alpha-blockers, ACE inhibitors, angiotensin II receptor blockers (ARBs), direct renin inhibitors, calcium channel blockers, central agonists, peripheral adrenergic blockers, vasodilators, insulin, alpha-glucosidase inhibitors, biguanides, dopamine agonists, DPP-4 inhibitors, glucagon-like peptides, meglitinides, sodium glucose transporter (SGLT) inhibitors, sulfonylureas, thiazolidinediones, pramlintide, statins, fibrates, aspirin and anticoagulants.

7. The use according to claim 1, wherein the human FGF21 protein variant is provided in the form of subcutaneous administration.

8. The use according to claim 1, wherein the solution contains 100 mg / mL to 150 mg / mL of human FGF21 protein variant.

9. The use according to claim 1, wherein the solution comprises a tromethamine buffer at a concentration of 10 mM to 50 mM.

10. The use according to claim 9, wherein the solution comprises tromethamine buffer at a concentration of 30 mM.

11. The use according to claim 1, wherein the solution comprises 250 mM to 300 mM sucrose.

12. Use according to claim 11, wherein the solution comprises 270 mM sucrose.

13. The use according to claim 1, wherein the solution comprises 0.02% to 0.10% polysorbate 20.

14. Use according to claim 13, wherein the solution comprises 0.06% polysorbate 20.

15. The use according to claim 1, wherein the pH value of the solution is in the range of 6.5 to 9.

16. The use according to claim 1, wherein the solution comprises about 30 mM tromethamine buffer, 270 mM sucrose and 0.06% polysorbate 20, pH 7.5 to 8.

5.

17. The use according to claim 15 or 16, wherein the pH of the solution is 8.

0.

18. A pharmaceutical composition comprising a human FGF21 protein variant, the pharmaceutical composition being a lyophilized preparation for reconstitution into a solution before administration, wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11, wherein the solution comprises 50 mg / mL to 150 mg / mL of the human FGF21 protein variant, tromethamine buffer, sucrose and polysorbate 20.

19. A pharmaceutical composition comprising a human FGF21 protein variant, wherein the pharmaceutical composition is a solution, wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11, wherein the solution comprises 50 mg / mL to 150 mg / mL of the human FGF21 protein variant, tromethamine buffer, sucrose and polysorbate 20.

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