Endothelial lipase antibodies for treatment of cardiovascular diseases

Abstract

To provide methods of using anti-EL antibodies and antibody-fragments thereof to effectively treat diseases and disorders, e.g., CV diseases and disorders.SOLUTION: Provided is a method of treating cardiovascular disease in a subject, the method comprising administering to the subject about 100 mg to about 350 mg of an antibody or antigen- binding fragment thereof that specifically binds to human endothelial lipase (EL).SELECTED DRAWING: None

Description

[Technical Field]

[0001] [CROSS-REFERENCE TO RELATED APPLICATIONS] This application claims priority to U.S. Provisional Application No. 63 / 104,410, filed October 22, 2020, U.S. Provisional Application No. 62 / 940,164, filed November 25, 2019, and U.S. Provisional Application No. 62 / 932,257, filed November 7, 2019, the entire disclosures of which are incorporated herein by reference.

[0002] 1. Field The present disclosure relates generally to methods of using antibodies and antigen-binding fragments thereof that specifically bind to human endothelial lipase (EL) for the treatment of diseases or disorders, such as cardiovascular diseases and disorders. Advantageous uses are provided. [Background technology]

[0003] 2.Background Endothelial lipase (EL) is a circulating phospholipase identified as a member of the triglyceride lipase family. EL possesses both phospholipase and triglyceride lipase activities and hydrolyzes high-density lipoproteins (HDL) more efficiently than other lipoproteins. EL is thought to play an important role in regulating plasma HDL cholesterol (HDL-C) levels. EL hydrolyzes HDL phospholipids, leading to HDL particle destabilization and rapid clearance by the kidney.

[0004] Increased plasma EL concentrations are associated with a worsening lipoprotein-lipid profile, along with elevated plasma triglyceride and apolipoprotein B concentrations, as well as smaller low-density lipoprotein particle sizes (Non-Patent Document 1). Increased proinflammatory cytokine concentrations and an increased prevalence of metabolic syndrome have also been observed among individuals with elevated plasma EL concentrations (Non-Patent Document 1). Considering these and other factors, EL is thought to play an important role in cardiovascular disease (Non-Patent Document 1).

[0005] Despite the effectiveness of current therapies for cardiovascular disease, such as high-potency statins, there remains a significant residual risk of major cardiovascular (CV) events in patients with acute coronary syndrome (ACS). Most myocardial infarctions (MIs) occur in patients with normal low-density lipoprotein (LDL) levels, and despite treatment with high-dose high-potency statins, PCSK9 inhibitors, and / or ezetimibe after an MI, the residual risk of a second CV event remains high. For example, in the IMPROVE-IT study (ezetimibe + statin), there was a 33% risk of CV events over 7 years of follow-up. Furthermore, in ODYSSEY (PCSK9 inhibitor + statin), there was a 9.5% residual risk over 2.8 years.

[0006] Low HDL-C levels have been identified as a predictor of atherosclerotic CV events and a risk factor for coronary heart disease (CHD). It has also been hypothesized that HDL particle size and particle number may be useful clinical markers of HDL and related diseases.

[0007] Several attempts have been made to pharmacologically raise HDL levels using various mechanisms of action. In particular, four trials of cholesterol transfer protein (CETP) inhibitors have been completed. While CETP inhibitors raise HDL cholesterol, three of the four trials did not improve CV outcomes, and one trial showed a modest reduction in CV events, resulting in only a 9% relative risk reduction. This approach has been criticized because CETP inhibition results in a blockade of LDL receptor-mediated reverse cholesterol transport.

[0008] Humans with partial and complete loss-of-function mutations in the gene encoding EL exhibit elevated HDL-C, increased cholesterol efflux capacity (CEC), and a tendency toward reduced CV risk. Therefore, neutralization of EL represents a promising therapeutic mechanism. However, there are currently no approved therapies that target EL or sufficiently reduce CV risk. Therefore, there is a need for methods of effectively treating diseases and disorders, such as CV diseases and disorders, using anti-EL antibodies and antibody fragments thereof. [Prior art documents] [Non-patent literature]

[0009] [Non-Patent Document 1] Paradis et al., Can J Cardiol 22: 31B-34B (2006) Summary of the Invention

[0010] 3. Overview Provided herein are methods for treating cardiovascular disease in a subject. In certain embodiments, the methods comprise administering to the subject about 100 mg to about 350 mg of an antibody or antigen-binding fragment thereof that specifically binds to human endothelial lipase (EL).

[0011] Provided herein are methods for reducing atherosclerosis in a subject. In certain embodiments, the methods comprise administering to the subject about 100 mg to about 350 mg of an antibody or antigen-binding fragment thereof that specifically binds to human EL.

[0012] Provided herein are methods for treating cardiovascular disease or reducing atherosclerosis in a subject. In certain embodiments, the methods comprise administering to the subject an antibody or antigen-binding fragment thereof that specifically binds EL, wherein administration of the antibody or antigen-binding fragment thereof (a) increases high-density lipoprotein cholesterol (HDL-C) in the subject, (b) increases high-density lipoprotein (HDL) particle number in the subject, (c) increases HDL particle size in the subject, and (d) increases IL-1 in the subject. (e) increase HDL phospholipids in a subject, (e) increase ApoA1 in a subject, and / or (f) increase cholesterol efflux capacity (CEC) in a subject. In certain embodiments, administration reduces the risk of cardiovascular death, non-fatal myocardial infarction (MI), non-fatal stroke, and / or coronary revascularization in a subject with a history of acute coronary syndrome (ACS). In certain embodiments, administration prevents a secondary cardiovascular event in a subject. In certain embodiments, administration reduces the risk of major adverse cardiovascular events (MACE) in a subject.

[0013] Provided herein are methods for reducing the risk of cardiovascular death, non-fatal myocardial infarction (MI), non-fatal stroke, and / or coronary revascularization in a subject with a history of acute coronary syndrome (ACS). In certain embodiments, the methods comprise administering to the subject an antibody or antigen-binding fragment thereof that specifically binds to human EL.

[0014] Provided herein are methods for preventing a secondary cardiovascular event in a subject. In certain embodiments, the methods comprise administering to the subject an antibody or antigen-binding fragment thereof that specifically binds to human EL.

[0015] Provided herein are methods for reducing the risk of major adverse cardiovascular events (MACE) in a subject. In certain embodiments, the methods comprise administering to the subject an antibody or antigen-binding fragment thereof that specifically binds to human EL.

[0016] In certain embodiments of the present disclosure, administration of the antibody or antigen-binding fragment thereof (a) increases high density lipoprotein cholesterol (HDL-C) in a subject, (b) increases high density lipoprotein (HDL) particle number in a subject, (c) increases HDL particle size in a subject, (d) increases HDL phospholipids in a subject, (e) increases ApoA1 in a subject, and / or (f) increases cholesterol efflux capacity (CEC) in a subject.

[0017] Provided herein are methods for increasing HDL-C, HDL particle number, HDL particle size, HDL phospholipids, ApoA1, and / or CEC in a subject. In certain embodiments, the methods comprise administering to the subject an antibody or antigen-binding fragment thereof that specifically binds to EL.

[0018] Certain embodiments of the present disclosure include administering about 100 mg to about 350 mg of an antibody or antigen-binding fragment thereof, such as about 100 mg, about 110 mg, about 120 mg, about 125 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 175 mg, about 180 mg, about 190 mg, about 200 mg, about 210 mg, about 220 mg, about 225 mg, about 230 mg, about 240 mg, about 250 mg, about 260 mg, about 270 mg, about 275 mg, about 280 mg, about 290 mg, about 300 mg, about 310 mg, about 320 mg, about 325 mg, about 330 mg, about 340 mg, or about 350 mg of an antibody or antigen-binding fragment thereof. Certain embodiments of the present disclosure include administering about 125 mg of an antibody or antigen-binding fragment thereof. Certain embodiments of the present disclosure include administering 125 mg of an antibody or antigen-binding fragment thereof. Certain embodiments of the present disclosure include administering about 250 mg of an antibody or antigen-binding fragment thereof. Certain embodiments of the present disclosure include administering 250 mg of an antibody or antigen-binding fragment thereof. Certain embodiments of the present disclosure include administering about 200 mg of an antibody or antigen-binding fragment thereof. Certain embodiments of the present disclosure include administering 200 mg to 250 mg of an antibody or antigen-binding fragment thereof.

[0019] In certain embodiments of the present disclosure, the antibody or antigen-binding fragment thereof is administered once a month. In certain embodiments of the present disclosure, the antibody or antigen-binding fragment thereof is administered once a month for at least 3 months. In certain embodiments of the present disclosure, the antibody or antigen-binding fragment thereof is administered once a month for at least 12 months or at least 24 months.

[0020] In certain embodiments of the present disclosure, the antibody or antigen-binding fragment thereof is administered parenterally. In certain embodiments of the present disclosure, the antibody or antigen-binding fragment thereof is administered subcutaneously.

[0021] In certain embodiments of the present disclosure, the antibody or antigen-binding fragment thereof is administered via an attached pre-filled syringe (APFS) or auto-injector.

[0022] In certain embodiments of the present disclosure, administration of the antibody or antigen-binding fragment thereof inhibits EL in a subject for 30 days.

[0023] In certain embodiments of the present disclosure, administration of the antibody or antigen-binding fragment thereof increases HDL-C in a subject by at least 30%. In certain embodiments of the present disclosure, administration of the antibody or antigen-binding fragment thereof increases HDL-C in a subject by at least 35%. In certain embodiments of the present disclosure, administration of the antibody or antigen-binding fragment thereof increases HDL-C in a subject by at least 40%. In certain embodiments of the present disclosure, administration of the antibody or antigen-binding fragment thereof increases HDL-C in a subject within 30 days of the first administration. In certain embodiments of the present disclosure, administration of the antibody or antigen-binding fragment thereof increases HDL-C in a subject within 90 days of the first administration.

[0024] In certain embodiments of the present disclosure, administration of the antibody or antigen-binding fragment thereof increases ApoA1 in a subject by at least 30%. In certain embodiments of the present disclosure, administration of the antibody or antigen-binding fragment thereof increases ApoA1 in a subject by at least 35%. In certain embodiments of the present disclosure, administration of the antibody or antigen-binding fragment thereof increases ApoA1 in a subject within 30 days of the first administration. In certain embodiments of the present disclosure, administration of the antibody or antigen-binding fragment thereof increases ApoA1 in a subject within 90 days of the first administration.

[0025] In certain embodiments of the present disclosure, administration of the antibody or antigen-binding fragment thereof increases non-ABCA1 cholesterol efflux capacity in a subject by at least 30%. In certain embodiments of the present disclosure, administration of the antibody or antigen-binding fragment thereof increases non-ABCA1 cholesterol efflux capacity in a subject by at least 35%. In certain embodiments of the present disclosure, administration of the antibody or antigen-binding fragment thereof increases non-ABCA1 cholesterol efflux capacity in a subject within 30 days of the first administration. In certain embodiments of the present disclosure, administration of the antibody or antigen-binding fragment thereof increases non-ABCA1 cholesterol efflux capacity in a subject within 90 days of the first administration.

[0026] In certain embodiments of the present disclosure, administration of the antibody or antigen-binding fragment thereof increases the number of HDL particles in a subject by at least 5% (e.g., as measured using NMR). In certain embodiments of the present disclosure, administration of the antibody or antigen-binding fragment thereof increases the number of HDL particles in a subject by at least 8% (e.g., as measured using NMR). In certain embodiments of the present disclosure, administration of the antibody or antigen-binding fragment thereof increases the number of HDL particles in a subject within 30 days of the first administration. In certain embodiments of the present disclosure, administration of the antibody or antigen-binding fragment thereof increases the number of HDL particles in a subject within 90 days of the first administration.

[0027] In certain embodiments of the present disclosure, administration of the antibody or antigen-binding fragment thereof increases HDL particle size in a subject by at least 3%. In certain embodiments of the present disclosure, administration of the antibody or antigen-binding fragment thereof increases HDL particle size in a subject by at least 5%. In certain embodiments of the present disclosure, administration of the antibody or antigen-binding fragment thereof increases HDL particle size in a subject within 30 days of the first administration. In certain embodiments of the present disclosure, administration of the antibody or antigen-binding fragment thereof increases HDL particle size in a subject within 90 days of the first administration.

[0028] In certain embodiments of the present disclosure, administration of the antibody or antigen-binding fragment thereof increases HDL phospholipids in a subject by at least 50%. In certain embodiments of the present disclosure, administration of the antibody or antigen-binding fragment thereof increases HDL phospholipids in a subject within 30 days of the first administration. In certain embodiments of the present disclosure, administration of the antibody or antigen-binding fragment thereof increases HDL phospholipids in a subject within 90 days of the first administration.

[0029] In certain embodiments of the disclosure, administration of the antibody or antigen-binding fragment thereof increases plasma phosphatidylinositol (PI) levels in the subject by at least 100% or at least 250%. In certain embodiments of the present disclosure, the increase in plasma PI levels is determined by the presence of a PI(14:2 / 20:0) level, a PI(14:2 / 22:0) level, a PI(14:2 / 22:1) level, a PI(14:2 / 22:2) level, a PI(16:0 / 16:1) level, a PI(16:0 / 18:0) level, a PI(16:0 / 18:2) level, a PI(16:0 / 20:2) level, a PI(16:0 / 20:3) level, a PI(16:0 / 20:4) level, a PI(16:0 / 22:4) level, a PI(16:1 / 18:0) level, a PI(16:1 / 18:1) level, a PI(18:0 / 18:0) level, a PI(18:0 / and / or PI(18:2 / 18:2) levels. In certain embodiments of the present disclosure, administration of the antibody or antigen-binding fragment thereof increases plasma phosphatidylinositol (PI) levels in the subject within 90 days of the first administration.

[0030] In certain embodiments of the present disclosure, the subject suffers from cardiovascular disease. In certain embodiments of the present disclosure, the cardiovascular disease is coronary artery disease, coronary heart disease (CHD), chronic arterial disease, cerebrovascular disease, atherosclerotic cardiovascular disease, or peripheral arterial disease.

[0031] In certain embodiments of the present disclosure, the subject has stable coronary artery disease or stable coronary heart disease. In certain embodiments of the present disclosure, the subject has a history of acute coronary syndrome (ACS).

[0032] In certain embodiments of the present disclosure, the subject is on statin therapy. In certain embodiments of the present disclosure, the subject is not on statin therapy.

[0033] In certain embodiments of the present disclosure, the subject has a triglyceride level of 500 mg / dL or less prior to administration. In certain embodiments of the present disclosure, the subject has an LDL-C of 100 mg / dL or less prior to administration.

[0034] In certain aspects of the present disclosure, the subject is a human.

[0035] In certain embodiments of the present disclosure, the antibody or antigen-binding fragment thereof neutralizes EL activity.

[0036] In certain embodiments of the present disclosure, the antibody or antigen-binding fragment thereof has reduced effector function. In certain embodiments of the present disclosure, the antibody or antigen-binding fragment thereof has no antibody-dependent cell-mediated cytotoxicity (ADCC) activity. In certain embodiments of the present disclosure, the antibody has no complement-dependent cytotoxicity (CDC) activity.

[0037] In certain embodiments of the present disclosure, the antibody binds to cynomolgus monkey EL.

[0038] In certain embodiments of the present disclosure, the antibody or antigen-binding fragment thereof competitively inhibits the binding to EL of an antibody comprising a VH comprising the amino acid sequence set forth in SEQ ID NO: 7 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 8. In certain embodiments of the present disclosure, the antibody or antigen-binding fragment thereof binds to the same epitope of EL as an antibody comprising a VH comprising the amino acid sequence set forth in SEQ ID NO: 7 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 8.

[0039] In certain embodiments of the present disclosure, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) complementarity-determining region (CDR)1, VH CDR2, VH CDR3, and a light chain variable region (VL) CDR1, VL CDR2, and VL CDR3 of the sequence of MEDI5884. In certain embodiments of the present disclosure, the CDRs are CDRs according to the Kabat definition, CDRs according to the Chothia definition, or CDRs according to the AbM definition. In certain embodiments of the present disclosure, the antibody or antigen-binding fragment thereof comprises a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 3, a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a VL CDR3 comprising the amino acid sequence of SEQ ID NO: 6.

[0040] In certain embodiments of the present disclosure, the antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO:7, and / or a VL comprising the amino acid sequence set forth in SEQ ID NO:8.

[0041] In certain embodiments of the present disclosure, the antibody or antigen-binding fragment thereof comprises an IgG heavy chain constant region. In certain embodiments of the present disclosure, the IgG heavy chain constant region is an IgG4 heavy chain constant region. In certain embodiments of the present disclosure, the IgG4 heavy chain constant region is an IgG4P heavy chain constant region. In certain embodiments of the present disclosure, the antibody or antigen-binding fragment thereof comprises a kappa light chain constant region.

[0042] In certain embodiments of the present disclosure, the antibody or antigen-binding fragment comprises a heavy chain constant region and / or a light chain constant region, hi certain embodiments of the present disclosure, the heavy chain constant region is a human IgG4P heavy chain constant region and / or the light chain constant region is a human IgGκ light chain constant region.

[0043] In certain aspects of the present disclosure, the antibody or antigen-binding fragment thereof is a humanized antibody or antigen-binding fragment thereof.

[0044] In certain embodiments of the present disclosure, the antibody or antigen-binding fragment thereof is selected from the group consisting of SEQ ID NO:1 2, and / or a light chain constant region comprising the amino acid sequence set forth in SEQ ID NO: 13.

[0045] In certain embodiments of the present disclosure, the antibody comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:9 and a light chain comprising the amino acid sequence set forth in SEQ ID NO:10.

[0046] In certain aspects of the present disclosure, the antibody or antigen-binding fragment thereof is a full-length antibody.

[0047] In certain embodiments of the present disclosure, the antibody or antigen-binding fragment thereof is an antigen-binding fragment, such as a Fab, Fab', F(ab')2, single-chain Fv (scFv), disulfide-linked Fv, V-NAR domain, IgNar, intrabody, IgGΔCH2, minibody, F(ab')3, tetrabody, triabody, diabody, single domain antibody, DVD-Ig, Fcab, or mAb. 2 , (scFv)2, or scFv-Fc.

[0048] Provided herein are methods for treating cardiovascular disease in a subject. In certain embodiments, the method comprises subcutaneously administering 250 mg of an antibody or antigen-binding fragment thereof to the subject once per month, wherein the antibody or antigen-binding fragment specifically binds to human EL and comprises a VH comprising the amino acid sequence set forth in SEQ ID NO:7 and a VL comprising the amino acid sequence set forth in SEQ ID NO:8. In certain embodiments, the method comprises subcutaneously administering about 200 mg of an antibody or antigen-binding fragment thereof to the subject once per month, wherein the antibody or antigen-binding fragment specifically binds to human EL and comprises a VH comprising the amino acid sequence set forth in SEQ ID NO:7 and a VL comprising the amino acid sequence set forth in SEQ ID NO:8. In certain embodiments, the method comprises subcutaneously administering 200 mg to 250 mg of an antibody or antigen-binding fragment thereof to the subject once per month, wherein the antibody or antigen-binding fragment specifically binds to human EL and comprises a VH comprising the amino acid sequence set forth in SEQ ID NO:7 and a VL comprising the amino acid sequence set forth in SEQ ID NO:8. Provided herein are methods for reducing the risk of cardiovascular death, non-fatal myocardial infarction (MI), non-fatal stroke, and / or coronary revascularization in subjects with a history of acute coronary syndrome (ACS). In certain embodiments, the method comprises subcutaneously administering 250 mg of an antibody or antigen-binding fragment thereof once per month to a subject, wherein the antibody or antigen-binding fragment specifically binds to human EL and comprises a VH comprising the amino acid sequence set forth in SEQ ID NO:7 and a VL comprising the amino acid sequence set forth in SEQ ID NO:8. In certain embodiments, the method comprises subcutaneously administering about 200 mg of an antibody or antigen-binding fragment thereof once per month to a subject, wherein the antibody or antigen-binding fragment specifically binds to human EL and comprises a VH comprising the amino acid sequence set forth in SEQ ID NO:7 and a VL comprising the amino acid sequence set forth in SEQ ID NO:8.In certain embodiments, the method comprises subcutaneously administering 200 mg to 250 mg of an antibody or antigen-binding fragment thereof to a subject once per month, wherein the antibody or antigen-binding fragment specifically binds to human EL and comprises a VH comprising the amino acid sequence set forth in SEQ ID NO:7 and a VL comprising the amino acid sequence set forth in SEQ ID NO:8.

[0049] In certain embodiments of the present disclosure, the antibody comprises the amino acid sequence of a heavy chain constant region set forth in SEQ ID NO:9 and the amino acid sequence of a light chain constant region set forth in SEQ ID NO:10.

[0050] In certain embodiments of the present disclosure, the method further comprises administering an inhibitor of proprotein convertase subtilisin / kexin type 9 (PCSK9). In certain embodiments, the administration of the antibody or antigen-binding fragment thereof that specifically binds to human EL and the administration of the inhibitor of PCSK9 are simultaneous. In certain embodiments, the administration of the antibody or antigen-binding fragment thereof that specifically binds to human EL is concurrent. The antibody or antigen-binding fragment thereof that specifically binds to human EL and the inhibitor of PCSK9 are administered in separate pharmaceutical compositions. In certain embodiments, the administration of the antibody or antigen-binding fragment thereof that specifically binds to human EL and the administration of the inhibitor of PCSK9 are sequential.

[0051] In certain embodiments, the inhibitor of PCSK9 is an anti-PCSK9 antibody or antigen-binding fragment thereof. In certain embodiments, the inhibitor of PCSK9 is HS9, evolocumab, alirocumab, or bococizumab.

[0052] 4. Brief description of the drawings [Brief explanation of the drawings]

[0053] [Figure 1A] FIG. 1 shows a dose-dependent increase in exposure to MEDI5584 (see Examples 4 and 5). [Figure 1B] FIG. 1 shows MEDI5884 dose-dependent target association (inhibition of EL) (see Examples 4 and 5). [Figure 2] FIG. 1 shows MEDI5884 dose-dependent increase in high density lipoprotein cholesterol (HDL-C) (see Examples 4 and 5). [Figure 3] FIG. 1 shows MEDI5884 dose-dependent increase in apolipoprotein A1 (ApoA1) (see Examples 4 and 5). [Figure 4] FIG. 1 shows MEDI5884 dose-dependent increase in high density lipoprotein phospholipids (HDL-PL) (see Examples 4 and 5). [Figure 5] FIG. 1 shows MEDI5884 dose-dependent increase in non-ATP-binding cassette transporter A1 (ABCA1) cholesterol efflux (see Example 4). [Figure 6] FIG. 1 shows the dose-response relationships of HDL-C, ApoA1, and HDL-PL based on the area under the effect curve between days 60 and 90 (AUECd60-90) (see Example 5). [Figure 7] Figure 1 shows pharmacokinetic (PK) and pharmacodynamic (PD) modeling for MEDI5884. CLd = intercompartmental clearance; CL = apparent total body clearance; conc = concentration; dHDL(t) / dt = change in HDL over time; HDL = high density lipoprotein cholesterol; HDL(t) = HDL level at time t; IC = concentration at which 50% of the maximum effect is reached (estimated simultaneously with K); IH = inhibitory effect from MEDI5884; I = maximum inhibitory effect; Ka = absorption rate; Kin = rate of HDL production; K = concentration at which 50% of V is reached; K = rate of HDL elimination; SC = subcutaneous; V = maximum contribution of dose-dependent nonlinear clearance (see Example 5). [Figure 8] FIG. 1 shows a PK simulation of MEDI5884 administered at 250 mg monthly (see Example 5). [Figure 9] FIG. 1 shows the study design analyzing the effect of combined inhibition of EL and proprotein convertase subtilisin / kexin type 9 (PCSK9) in cynomolgus monkeys. [Figure 10] 1 shows the effect of combined inhibition of EL and PCSK9 on LDL-C and HDL-C levels in cynomolgus monkeys. Data are expressed as the mean change ± SEM from baseline (day 0) measurements. In each plot, the mean baseline value (n=16) is shown (see Example 6). [Figure 11]

[0023] Figure 1 shows the effect of combined inhibition of EL and PCSK9 on ApoB and ApoA1 levels in cynomolgus monkeys. Data are expressed as the mean change ± SEM from baseline (day 0) measurements. In each plot, the mean baseline value (n = 16) is shown (see Example 6). [Figure 12] FIG. 1 shows the effect of combined inhibition of EL and PCSK9 on cholesterol efflux capacity (total efflux and ABCA1 efflux) in cynomolgus monkeys (see Example 6). [Figures 13A-13C] FIG. 1 shows the time course and dose-response of plasma phosphatidylinositol (PI) species for selected doses in a single ascending dose (SAD) and multiple ascending dose (MAD) study in healthy patients and patients with coronary artery disease (CAD) (see Example 7). [Figures 14A-14C] FIG. 1 shows the time course and dose-response of plasma phosphatidylinositol (PI) species for all doses in a single ascending dose (SAD) and multiple ascending dose (MAD) study in healthy patients and patients with coronary artery disease (CAD) (see Example 7). [Figure 15] FIG. 1 shows the effect of MEDI5884 on plasma phosphatidylinositol (PI) species in healthy volunteers and CAD patients at day 21 (see Example 7). [Figure 16] FIG. 1 shows the effect of MEDI5884 on plasma phosphatidylinositol (PI) species in healthy volunteers and patients with coronary artery disease (CAD) throughout the day (see Example 7). [Figures 17A-17E]FIG. 1 shows the percent change from baseline for various plasma phosphatidylinositol (PI) species in patients with coronary artery disease (CAD) treated with various doses of MEDI5884 or placebo (see Example 7). [Figure 18] FIG. 1 shows the mean percent change from baseline for all measured plasma phosphatidylinositol (PI) species in coronary artery disease (CAD) patients treated with various doses of MEDI5884 or placebo (see Example 7). [Figures 19A-19E] FIG. 1 shows the percent change from baseline for various plasma phosphatidylinositol (PI) species in healthy volunteers treated with various doses of MEDI5884 or placebo (see Example 7). DETAILED DESCRIPTION OF THE INVENTION

[0054] 5. Detailed Description Provided herein are methods for administering antibodies (e.g., monoclonal antibodies) and antigen-binding fragments thereof that specifically bind to endothelial lipase (EL, e.g., human EL). Anti-EL antibodies and antigen-binding fragments thereof can be administered to, for example, treat cardiovascular disease in a subject. Anti-EL antibodies or antigen-binding fragments thereof can increase high-density lipoprotein cholesterol (HDL-C), increase HDL particle number, increase HDL particle size, increase HDL phospholipids, increase ApoA1, and / or increase cholesterol efflux capacity in a subject. In some embodiments of the present disclosure, about 100 mg to about 350 mg (e.g., about 250 mg) of the antibody or antigen-binding fragment thereof is administered to a subject, for example, about once per month (QM).

[0055] 5.1 Terminology As used herein, the term "endothelial lipase" or "EL" refers to mammalian EL polypeptides, including, but not limited to, naturally occurring EL polypeptides and isoforms of EL polypeptides. "EL" encompasses not only full-length, unprocessed EL polypeptides, but also forms of EL polypeptides that result from intracellular processing. As used herein, the term "human EL" refers to a polypeptide comprising the amino acid sequence of SEQ ID NO: 11. "EL polynucleotide," "EL nucleotide," or "EL nucleic acid" refers to a polynucleotide encoding EL.

[0056] The term "antibody" refers to an immunoglobulin molecule that recognizes and specifically binds to a target, such as a protein, polypeptide, peptide, carbohydrate, polynucleotide, lipid, or a combination thereof, via at least one antigen recognition site in the variable region of the immunoglobulin molecule. As used herein, the term "antibody" refers to an intact polyclonal antibody, an intact monoclonal antibody, or an intact polyclonal antibody, as long as the antibody exhibits the desired biological activity. Antibodies include monoclonal antibodies, chimeric antibodies, humanized antibodies, human antibodies, antibody-containing fusion proteins, and any other modified immunoglobulin molecules. Antibodies can be any of the five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, or their subclasses (isotypes), designated alpha, delta, epsilon, gamma, and mu, based on the identity of their heavy chain constant domains (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2). Different classes of immunoglobulins have distinct and well-known subunit structures and three-dimensional configurations. Antibodies can be naked or conjugated to other molecules such as toxins, radioisotopes, etc.

[0057] The term "antibody fragment" refers to a portion of an intact antibody. An "antigen-binding fragment," "antigen-binding domain," or "antigen-binding region" refers to the portion of an intact antibody that binds to an antigen. An antigen-binding fragment may contain the antigen recognition site of the intact antibody (e.g., a complementarity-determining region (CDR) sufficient to specifically bind to the antigen). Examples of antigen-binding fragments of antibodies include, but are not limited to, Fab fragments, Fab' fragments, F(ab')2 fragments, and Fv fragments, linear antibodies, and single-chain antibodies. Antigen-binding fragments of antibodies The antibodies may be derived from any animal species, including rodents (e.g., mouse, rat, or hamster), and humans, or may be artificially produced.

[0058] The terms "anti-EL antibody," "EL antibody," and "antibody that binds to EL" refer to an antibody that can specifically bind to EL with sufficient affinity such that the antibody is useful as a diagnostic and / or therapeutic agent in targeting EL. As used herein, the terms "specifically bind," "immunospecifically bind," "immunospecifically recognize," and "specifically recognize" are similar terms in the context of an antibody or antigen-binding fragment thereof. These terms indicate that the antibody or antigen-binding fragment thereof binds to an epitope via its antigen-binding domain and that the binding involves some complementarity between the antigen-binding domain and the epitope. Thus, an antibody that "specifically binds" to human EL (SEQ ID NO: 11) may also bind to EL from other species (e.g., cynomolgus monkeys) and / or EL proteins produced from other human alleles, but the extent of binding to unrelated non-EL proteins (e.g., other lipases, such as hepatic lipase or lipoprotein lipase) is less than about 10% of the antibody's binding to EL, as measured, for example, by an in vitro neutralization assay.

[0059] A "monoclonal" antibody or antigen-binding fragment thereof refers to a homogeneous population of antibodies or antigen-binding fragments that are involved in highly specific binding of a single antigenic determinant or epitope. This is in contrast to polyclonal antibodies, which typically include different antibodies directed against different antigenic determinants. The term "monoclonal" antibody or antigen-binding fragment thereof encompasses both intact and full-length monoclonal antibodies, as well as antibody fragments (Fab, Fab', F(ab')2, Fv, etc.), single-chain (scFv) variants, fusion proteins containing an antibody portion, and any other modified immunoglobulin molecule containing an antigen-recognition site. Furthermore, a "monoclonal" antibody or antigen-binding fragment thereof refers to such antibodies and antigen-binding fragments thereof produced by any number of methods, including, but not limited to, hybridoma, phage selection, recombinant expression, and transgenic animals.

[0060] As used herein, the terms "variable region" and "variable domain" are used interchangeably and are common in the art. The variable region typically refers to a portion of an antibody, generally a portion of either the light or heavy chain, typically the amino-terminal approximately 110-120 or 110-125 amino acids of the mature heavy chain and approximately 90-115 amino acids of the mature light chain, which differ in sequence between antibodies and are used to determine the binding and specificity of a particular antibody for a particular antigen. The sequence variability is concentrated in regions called complementarity-determining regions (CDRs), although variable regions may also be present. The more highly conserved regions within the variable domains are called framework regions (FRs). Without wishing to be bound by a particular mechanism or theory, it is believed that the CDRs of the light and heavy chains are primarily responsible for antibody-antigen interaction and specificity. In some embodiments of the present disclosure, the variable regions are human variable regions. In some embodiments of the present disclosure, the variable regions comprise rodent or murine CDRs and human framework regions (FRs). In certain embodiments of the present disclosure, the variable regions are primate (e.g., non-human primate) variable regions. In some embodiments of the present disclosure, the variable regions comprise rodent or murine CDRs and primate (e.g., non-human primate) framework regions (FRs).

[0061] The terms "VL" and "VL domain" are used interchangeably to refer to the light chain variable region of an antibody.

[0062] The terms "VH" and "VH domain" are used interchangeably to refer to the heavy chain variable region of an antibody.

[0063] The term "Kabat numbering" and similar terms are recognized in the art and refer to a system for numbering amino acid residues in the variable regions of the heavy and light chains of an antibody, or an antigen-binding fragment thereof. In some embodiments, CDRs can be identified according to the Kabat numbering system (see, e.g., Kabat EA & Wu TT (1971) Ann NY Acad Sci 190: 382-391, and Kabat EA et al., (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No. 91-3242). Using the Kabat numbering system, Using the Kabat numbering system, the CDRs in an antibody heavy chain molecule are typically located at amino acids 31-35 (CDR1), 50-65 (CDR2), and 95-102 (CDR3), which may include 35 followed by optionally one or two additional amino acids (referred to as 35A and 35B in the Kabat numbering scheme). Using the Kabat numbering system, the CDRs in an antibody light chain molecule are typically located at amino acids 24-34 (CDR1), 50-56 (CDR2), and 89-97 (CDR3). In some embodiments of the present disclosure, the CDRs of the antibodies described herein were determined according to the Kabat numbering scheme.

[0064] Instead, Chothia refers to the location of the structural loops (Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987)). The end of the Chothia CDR-H1 loop, when numbered using the Kabat numbering convention, varies between H32 and H34 depending on the length of the loop (this is because the Kabat numbering scheme places insertions at H35A and H35B; if neither 35A nor 35B is present, the loop ends at 32; if only 35A is present, the loop ends at 33; if both 35A and 35B are present, the loop ends at 34). The AbM hypervariable regions represent a compromise between the Kabat CDRs and the Chothia structural loops and are used in Oxford Molecular's AbM antibody modeling software.

[0065] TIFF2025131673000001.tif49170

[0066] As used herein, the terms "constant region" and "constant domain" are used interchangeably and have a common meaning in the art. The constant region is the portion of an antibody, e.g., the carboxyl-terminal portion of the light and / or heavy chain, that is not directly involved in binding the antibody to an antigen, but may exhibit various effector functions, such as interaction with Fc receptors. The constant region of an immunoglobulin molecule generally has a more conserved amino acid sequence compared to the immunoglobulin variable domain.

[0067] As used herein, the term "heavy chain," when used in reference to an antibody, can refer to any of the different types, e.g., alpha (α), delta (δ), epsilon (ε), gamma (γ), and mu (μ), which give rise to antibodies of the IgA, IgD, IgE, IgG, and IgM classes (which include IgG subclasses, e.g., IgG1, IgG2, IgG3, and IgG4), respectively, based on the amino acid sequence of the constant domain. Heavy chain amino acid sequences are well known in the art. In some embodiments of the present disclosure, the heavy chain is a human heavy chain.

[0068] As used herein, the term "light chain," when used in reference to an antibody, can refer to any of different types, such as kappa (κ) or lambda (λ), based on the amino acid sequence of the constant domain. Light chain amino acid sequences are well known in the art. In some embodiments of the present disclosure, the light chain is a human light chain.

[0069] The term "MEDI5884" refers to an anti-EL antibody comprising a heavy chain of SEQ ID NO: 9 and a light chain of SEQ ID NO: 10. MEDI5884 is also known as "S6F1-4P" and comprises the heavy chain variable region of the h55A1-S6 antibody and the light chain variable region of the h55A1-F1 antibody disclosed in U.S. Patent Application Publication No. 2017 / 0260290, which is incorporated herein by reference in its entirety.

[0070] The term "chimeric" antibody or antigen-binding fragment thereof refers to an antibody or antigen-binding fragment thereof whose amino acid sequences are derived from two or more species. Typically, the variable regions of both the light and heavy chains correspond to the variable regions of an antibody or antigen-binding fragment thereof from one mammalian species (e.g., mouse, rat, rabbit, etc.) with the desired specificity, affinity, and capacity, while the constant regions are homologous to the sequences of an antibody or antigen-binding fragment thereof from another species (usually human) to avoid eliciting an immune response in that species.

[0071] The term "humanized" antibody or antigen-binding fragment thereof refers to a form of a non-human (e.g., murine) antibody or antigen-binding fragment thereof that is a specific immunoglobulin chain, chimeric immunoglobulin, or fragment thereof that contains minimal non-human (e.g., murine) sequence. Typically, a humanized antibody or antigen-binding fragment thereof is one in which the remainder from the complementarity-determining regions (CDRs) is derived from a non-human species (e.g., murine, Humanized antibodies or antigen-binding fragments thereof are human immunoglobulins in which residues from the CDRs of non-human species (rat, rabbit, hamster) have been replaced ("CDR-grafted") (Jones et al., Nature 321:522-525 (1986); Riechmann et al., Nature 332:323-327 (1988); Verhoeyen et al., Science 239:1534-1536 (1988)). In some cases, certain Fv framework region (FR) residues of a human immunoglobulin are replaced with corresponding residues in an antibody or fragment from a non-human species having the desired specificity, affinity, and capacity. Humanized antibodies or antigen-binding fragments thereof can be further modified by substitution of additional residues either in the Fv framework regions and / or within the non-human CDR residues to refine and optimize the specificity, affinity, and / or capacity of the antibody or antigen-binding fragment thereof. Generally, a humanized antibody or antigen-binding fragment thereof will contain variable domains that include all or substantially all of the CDR regions corresponding to those of a non-human immunoglobulin, while all or substantially all of the FR regions are of the consensus sequence of a human immunoglobulin. The humanized antibody or antigen-binding fragment thereof may also contain at least a portion of an immunoglobulin constant region or domain (Fc), typically that of a human immunoglobulin. Examples of methods used to make humanized antibodies are described in U.S. Patent No. 5,225,539; Roguska et al., Proc. Natl. Acad. Sci., USA, 91(3):969-973 (1994); and Roguska et al., Protein Eng. 9(10):895-904 (1996). In some embodiments of the present disclosure, the "humanized antibody" is a resurfaced antibody.

[0072] The term "human" antibody or antigen-binding fragment thereof means an antibody or antigen-binding fragment thereof having an amino acid sequence derived from the human immunoglobulin locus, where such antibody or antigen-binding fragment is made using any technique known in the art. This definition of a human antibody or antigen-binding fragment thereof includes intact or full-length antibodies and fragments thereof.

[0073] "Binding affinity" generally refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., an antibody or antigen-binding fragment thereof) and its binding partner (e.g., an antigen). Unless otherwise indicated, as used herein, "binding affinity" refers to the intrinsic binding affinity, reflecting a 1:1 interaction between members of a binding pair (e.g., an antibody or antigen-binding fragment thereof and an antigen). The affinity of a molecule X for its partner Y is generally determined by the dissociation constant (K D Affinity can be expressed by, but not limited to, the equilibrium dissociation constant (K D ) and the equilibrium association constant (K A ) can be measured and / or expressed in a number of ways known in the art. D is k off / k on It is calculated from the quotient of K A is k on / k off It is calculated from the quotient of k on refers to the association rate constant of, for example, an antibody or antigen-binding fragment thereof to an antigen, and k off k refers to, for example, the dissociation of an antibody or antigen-binding fragment thereof from an antigen. on and k off can be determined by techniques known to those skilled in the art such as BIAcore™ or KinExA.

[0074] As used herein, "epitope" is a term used in the art and refers to a localized region of an antigen to which an antibody or antigen-binding fragment thereof can specifically bind. An epitope can be, for example, contiguous amino acids of a polypeptide (a linear or contiguous epitope), or an epitope can be, for example, derived from two or more non-contiguous regions of a polypeptide(s) together (a conformational, non-linear, discontinuous, or non-contiguous epitope). In some embodiments of the present disclosure, the epitope to which an antibody or antigen-binding fragment thereof specifically binds can be identified by, for example, NMR spectroscopy, X-ray diffraction crystallography studies, ELISA assays, hydrogen / deuterium exchange coupled with mass spectrometry (e.g., liquid chromatography electrospray mass spectrometry), array-based oligopeptide scanning assays, and / or mutagenesis mapping (e.g., site-directed mutagenesis mapping). For X-ray crystallography studies, crystallization can be performed using methods known in the art, e.g., For example, this can be accomplished using any of the methods described in Giege R et al. (1994) Acta Crystallogr D Biol Crystallogr 50(Pt 4): 339-350, McPherson A (1990) Eur J Biochem 189: 1-23, Chayen NE (1997) Structure 5: 1269-1274, and McPherson A (1976) J Biol Chem 251: 6300-6303. Antibody / antigen-binding fragment thereof:antigen crystals can be studied using well-known X-ray diffraction techniques, such as X-PLOR (Yale University, 1992, distributed by Molecular Simulations, Inc.; see, e.g., Meth Enzymol (1985) volumes 114 & 115, eds. Wyckoff, J. Am. Soc. Chem ... See HW et al., U.S. Patent Application Publication No. 2004 / 0014194), and Mutagenesis mapping studies can be performed using any method known to those skilled in the art. For example, see Champa M et al., (1995) J Biol Chem 270: 1388-1394, and Cunningham BC & Wells JA (1989) Science 244: 1081-1085 for a description of mutagenesis techniques, including alanine scanning mutagenesis.

[0075] An antibody that "binds to the same epitope" as a reference antibody refers to an antibody that binds to the same amino acid residue as the reference antibody. The ability of an antibody to bind to the same epitope as a reference antibody can be determined using a hydrogen / deuterium exchange assay (see Coales et al. Rapid Commun. Mass Spectrom. 2009; 23:639-647). This can be determined by X-ray crystallography.

[0076] An antibody is said to "competitively inhibit" the binding of a reference antibody to a given epitope if it preferentially binds to that epitope or an overlapping epitope to the extent that it blocks, to some extent, the binding of the reference antibody to the epitope. Competitive inhibition can be determined by any method known in the art, for example, by competitive ELISA assays. An antibody can be said to competitively inhibit the binding of the reference antibody to a given epitope by at least 90%, at least 80%, at least 70%, at least 60%, or at least 50%.

[0077] As used herein, an "inhibitor of PCSK9" is an agent that blocks the interaction of PCSK9 with the LDL receptor.

[0078] An "isolated" polypeptide, antibody, polynucleotide, vector, cell, or composition is a polypeptide, antibody, polynucleotide, vector, cell, or composition in a form not found in nature. Isolated polypeptides, antibodies, polynucleotides, vectors, cells, or compositions include those that have been purified to the extent that they are no longer in the form found in nature. In some embodiments of the present disclosure, an isolated antibody, polynucleotide, vector, cell, or composition is substantially pure. As used herein, "substantially pure" refers to a material that is at least 50% pure (i.e., free from contaminants), at least 90% pure, at least 95% pure, at least 98% pure, or at least 99% pure.

[0079] The terms "polypeptide," "peptide," and "protein" are used interchangeably herein to refer to polymers of amino acids of any length. Polymers can be linear or branched, can comprise modified amino acids, and can be interrupted by non-amino acids. The term also encompasses amino acid polymers that are modified, either naturally or by intervention (e.g., disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation or modification, such as conjugation with a labeling component). Also included within the definition are, for example, polypeptides that contain one or more analogs of an amino acid (including, for example, unnatural amino acids, etc.), as well as other modifications known in the art. Because the polypeptides of the present disclosure are based on antibodies, in some aspects of the present disclosure, the polypeptides are single-chain. It is understood that the amino acid sequence may be present as a single chain or as a related chain.

[0080] As used herein, the term "host cell" can be any type of cell, e.g., a primary cell, a cell in culture, or a cell from a cell line. In some embodiments of the present disclosure, the term "host cell" refers to a cell that has been transfected with a nucleic acid molecule and the progeny or potential progeny of such a cell. The progeny of such a cell may not be identical to the parent cell transfected with the nucleic acid molecule due, for example, to mutations or environmental influences that may occur in subsequent generations or upon integration of the nucleic acid molecule into the host cell genome.

[0081] The term "pharmaceutical formulation" refers to a formulation that is in a form that allows the biological activity of the active ingredient to be effective and that does not contain additional ingredients that are unacceptably toxic to the subject to which the formulation is administered. The formulation may be sterile.

[0082] The terms "administer," "administering," "administration," and the like, as used herein, refer to methods (e.g., intravenous administration) that can be used to enable delivery of a drug, such as an anti-EL antibody or antigen-binding fragment thereof, to a desired site of biological action. Administration techniques that can be used in the agents and methods described herein can be found, for example, in Goodman and Gilman, The Pharmacological Basis of Therapeutics, current edition, Pergamon; and Remington's, Pharmaceutical Sciences, current edition, Mack Publishing Co., Easton, Pa.

[0083] As used herein, the terms "subject" and "patient" are used interchangeably. A subject can be an animal. In some embodiments of the present disclosure, the subject is a mammal, such as a non-human animal (e.g., a cow, pig, horse, cat, dog, rat, mouse, monkey, or other primate, etc.). In some embodiments of the present disclosure, the subject is a cynomolgus monkey. In some embodiments of the present disclosure, the subject is a human.

[0084] The term "therapeutically effective amount" refers to an amount of a drug, e.g., an anti-EL antibody or antigen-binding fragment thereof, effective to treat a disease or disorder in a subject. Terms such as "treat," "treatment," "treating," "alleviate," and "alleviating" refer to therapeutic measures that cure, slow, reduce the symptoms of, and / or halt the progression of a pathological condition or disorder. Thus, subjects in need of treatment include those already diagnosed with or suspected of having the disorder. Administration "in combination with" one or more further therapeutic agents includes simultaneous (concurrent) or consecutive administration in any order.

[0085] As used in this disclosure and in the claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. nothing.

[0086] Whenever an aspect of the present disclosure is described herein with the word "comprising," it is understood that analogous aspects described in terms of "consisting of" and / or "consisting essentially of" are also provided.

[0087] As used herein, the term "or" is understood to be inclusive unless specifically stated otherwise or clear from the context. The term "and / or" when used herein in phrases such as "A and / or B" is intended to include both "A and B," "A or B," "A" and "B." Similarly, the term "and / or" when used in phrases such as "A, B, and / or C" is intended to include A, B, and C; A, B, or C; A or C; A or B; B, or C; A and C; A and B; B, and C; A (alone); It is intended to encompass each of the embodiments B (alone); and C (alone).

[0088] As used herein, the terms "about" and "approximately," when used to modify a numerical value or numerical range, indicate that deviations of 5% to 10% above and below that value or range are also within the intended meaning of the specified value or range.

[0089] Any composition or method provided herein can be combined with one or more of any of the other compositions and methods provided herein.

[0090] 5.2 Therapeutic Methods Using Anti-EL Antibodies or Antigen-Binding Fragments Thereof Provided herein are methods of administering an anti-EL antibody or antigen-binding fragment thereof described herein, or a pharmaceutical composition thereof described herein, to a subject in need thereof.

[0091] As provided herein, administration of an anti-EL antibody or antigen-binding fragment thereof, or a pharmaceutical composition thereof, can treat cardiovascular disease in a subject (e.g., a human subject). The cardiovascular disease can be, for example, coronary artery disease, coronary heart disease, cerebrovascular disease, or peripheral artery disease.

[0092] Administration of an anti-EL antibody or antigen-binding fragment thereof, or a pharmaceutical composition thereof, can reduce or prevent atherosclerosis in a subject (eg, a human subject).

[0093] Administration of an anti-EL antibody or antigen-binding fragment thereof, or a pharmaceutical composition thereof can prevent or reduce the risk of a secondary cardiovascular event in a subject (eg, a human subject).

[0094] Administration of an anti-EL antibody or antigen-binding fragment thereof, or a pharmaceutical composition thereof, can reduce the risk of cardiovascular death, non-fatal myocardial infarction (MI), non-fatal stroke, coronary revascularization, or any combination thereof in a subject (e.g., a human subject). The subject can be, for example, a subject with a history of acute coronary syndrome (ACS).

[0095] Administration of an anti-EL antibody or antigen-binding fragment thereof, or a pharmaceutical composition thereof can prevent or reduce the risk of a major adverse cardiovascular event (MACE) in a subject (eg, a human subject).

[0096] Administration of an anti-EL antibody or antigen-binding fragment thereof, or a pharmaceutical composition thereof, can (i) increase high density lipoprotein cholesterol (HDL-C), (ii) increase high density lipoprotein (HDL) particle number, (iii) increase HDL particle size, (iv) increase HDL phospholipids, (v) increase ApoA1, (vi) increase cholesterol efflux capacity (CEC), or (vii) any combination thereof.

[0097] Administration of an anti-EL antibody or antigen-binding fragment thereof, or a pharmaceutical composition thereof, can also increase plasma phosphatidylinositol (PI) levels.

[0098] In some embodiments, administration of an anti-EL antibody or antigen-binding fragment thereof inhibits EL.

[0099] In some embodiments, administration of an anti-EL antibody or antigen-binding fragment thereof induces a vasoconstriction in the subject. Administration of an anti-EL antibody or antigen-binding fragment thereof can increase HDL-C in the blood. Administration of an anti-EL antibody or antigen-binding fragment thereof can increase HDL-C, for example, by at least 30%, at least 35%, or at least 40%. Thus, administration of an anti-EL antibody or antigen-binding fragment thereof can increase HDL-C by about 30% to about 125%, about 30% to about 100%, about 30% to about 75%, about 30% to about 50%, about 30% to about 45%, or about 30% to about 40%. The increase in HDL-C can occur within 30 days of the first administration, within 60 days of the first administration, or within 90 days of the first administration.

[0100] In some embodiments, administration of an anti-EL antibody or antigen-binding fragment thereof increases the number of HDL particles in a subject. Administration of an anti-EL antibody or antigen-binding fragment thereof can increase HDL particle number by, for example, at least 5%, at least 8%, at least 10%, or at least 15% (e.g., as measured using NMR). Thus, administration of an anti-EL antibody or antigen-binding fragment thereof can increase HDL particle number by about 5% to about 20%, or about 5% to about 18% (e.g., as measured using NMR). The increase in HDL particle number can occur within 30 days of the first administration, within 60 days of the first administration, or within 90 days of the first administration.

[0101] In some embodiments, administration of an anti-EL antibody or antigen-binding fragment thereof increases HDL particle size in a subject. Administration of an anti-EL antibody or antigen-binding fragment thereof can increase HDL particle size, for example, by at least 3%, or at least 5%. Thus, administration of an anti-EL antibody or antigen-binding fragment thereof can increase HDL particle size by about 3% to about 10%, or about 3% to about 7%. The increase in HDL particle size can occur within 30 days of the first administration, within 60 days of the first administration, or within 90 days of the first administration.

[0102] In some embodiments, administration of an anti-EL antibody or antigen-binding fragment thereof increases HDL phospholipids in a subject. Administration of an anti-EL antibody or antigen-binding fragment thereof can increase HDL phospholipids, for example, by at least 50%. The increase in HDL phospholipids can occur within 30 days of the first administration, within 60 days of the first administration, or within 90 days of the first administration.

[0103] In some embodiments, administration of an anti-EL antibody or antigen-binding fragment thereof increases apolipoprotein A1 (apoA1) in a subject. Administration of an anti-EL antibody or antigen-binding fragment thereof can increase apoA1, for example, by at least 30%. Thus, administration of an anti-EL antibody or antigen-binding fragment thereof can increase apoA1 by about 30% to about 40%, or about 30% to about 35%. The increase in apoA1 can occur within 30 days of the first administration, within 60 days of the first administration, or within 90 days of the first administration.

[0104] In some embodiments, administration of an anti-EL antibody or antigen-binding fragment thereof increases cholesterol efflux capacity (CEC) in a subject. CEC can be measured, for example, using the method described in Thacker et al., Journal of Lipid Research 56: 1282-1295 (2015). Administration of an anti-EL antibody or antigen-binding fragment thereof can increase non-ATP-binding cassette transporter A1 (ABCA1) cholesterol efflux capacity, for example, by at least 30%, or at least 35%. Thus, administration of an anti-EL antibody or antigen-binding fragment thereof can increase non-ABCA1 cholesterol efflux capacity by about 30% to about 40%, or about 30% to about 35%.

[0105] In some embodiments, administration of an anti-EL antibody or antigen-binding fragment thereof increases plasma phosphatidylinositol (PI) levels in the subject. Therefore, the increase in PI levels was due to the following: PI(14:2 / 20:0) levels, PI(14:2 / 22:0) levels, PI(14:2 / 22:1) levels, PI(14:2 / 22:2) levels, PI(16:0 / 16:1) levels, PI(16:0 / 18:0) levels, PI(16:0 / 18:2) levels, PI(16:0 / 20:2) levels, PI(16:0 / 20:3) levels, PI(16:0 / 20:4) levels, PI(16:0 / 22:4) levels, PI(16:1 / 18:0) levels, PI(16:1 / 18:1) levels, PI(18:0 / 18:0) levels, PI(18:0 / 18:1) levels , increased PI(18:0 / 18:2) levels, PI(18:0 / 18:3) levels, PI(18:0 / 20:2) levels, PI(18:0 / 20:3) levels, PI(18:0 / 20:4) levels, PI(18:0 / 22:4) levels, PI(18:0 / 22:5) levels, PI(18:0 / 22:6) levels, PI(18:1 / 16:0) levels, PI(18:1 / 18:1) levels, PI(18:1 / 18:2) levels, PI(18:1 / 20:2) levels, PI(18:1 / 20:3) levels, PI(18:1 / 20:4) levels, and / or PI(18:2 / 18:2) levels. In some embodiments, the increase in PI levels is an increase in PI(14:2 / 22:2) levels, PI(16:0 / 16:1) levels, PI(16:0 / 18:2) levels, PI(16:0 / 20:3) levels, PI(16:0 / 20:4) levels, PI(18:0 / 18:1) levels, PI(18:0 / 18:2) levels, PI(18:0 / 20:2) levels, PI(18:0 / 20:3) levels, PI(18:0 / 20:4) levels, PI(18:0 / 22:6) levels, and / or PI(18:1 / 16:0) levels. Administration of an anti-EL antibody or antigen-binding fragment thereof can increase plasma PI levels, for example, by at least several hundred percent for more abundant PI species, and by several hundred to several thousand percent change from baseline for less abundant PI species. Thus, administration of anti-EL antibodies or antigen-binding fragments thereof can increase plasma PI by approximately 100% to 1000%, depending on the PI species.In some cases, administration of an anti-EL antibody or antibody-binding fragment thereof increases the levels of at least 10 plasma PI species, for example, by at least 100% or between 100% and 1000%. In some cases, administration of an anti-EL antibody or antigen-binding fragment thereof increases the levels of at least 12 plasma PI species (e.g., PI(14:2 / 22:2), PI(16:0 / 16:1), PI(16:0 / 18:2), PI(16:0 / 20:3), PI(16:0 / 20:4), PI(18:0 / 18:1), PI(18:0 / 18:2), PI(18:0 / 20:2), PI(18:0 / 20:3), PI(18:0 / 20:4), PI(18:0 / 22:6), and PI(18:1 / 16:0)), e.g., by at least 100%, or between 100% and 1000%. In some cases, administration of an anti-EL antibody or antigen-binding fragment thereof may be inducible by administration of at least 30 plasma PI species (e.g., PI(14:2 / 20:0), PI(14:2 / 22:0), PI(14:2 / 22:1), PI(14:2 / 22:2), PI(16:0 / 16:1), PI(16:0 / 18:0), PI(16:0 / 18:2), PI(16:0 / 20:2), PI(16:0 / 20:3), PI(16:0 / 20:4), PI(16:0 / 22:4), PI(16:1 / 18:0), PI(16:1 / 18:1), PI(18:0 / 18:0), PI(18:0 / 20:4), PI(16:0 / 22:4), PI(16:1 / 18:0), PI(16:1 / 18:1), PI(18:0 / 18:0), PI(18:0 / 20:2), PI(16:0 / 20:3), PI(16:0 / 20:4), PI(16:0 / 22:4), PI(16:1 / 18:0), PI(16:1 / 18:1), PI(18:0 / 18:0), PI(18:0 / 20:2), PI(16 ... The levels of PI(18:1), PI(18:0 / 18:2), PI(18:0 / 18:3), PI(18:0 / 20:2), PI(18:0 / 20:3), PI(18:0 / 20:4), PI(18:0 / 22:4), PI(18:0 / 22:5), PI(18:0 / 22:6), PI(18:1 / 16:0), PI(18:1 / 18:1), PI(18:1 / 18:2), PI(18:1 / 20:2), PI(18:1 / 20:3), PI(18:1 / 20:4), and PI(18:2 / 18:2) are increased, for example, by at least 100% or 100% to 1000%.

[0106] In some cases, administration of an anti-EL antibody or antigen-binding fragment thereof increases the levels of at least 10 plasma PI species by at least 250%, or between 250% and 1000%. In some cases, administration of an anti-EL antibody or antigen-binding fragment thereof increases the levels of at least 12 plasma PI species by at least 250%, or between 250% and 1000%. Increase levels of plasma PI species (e.g., PI(14:2 / 22:2), PI(16:0 / 16:1), PI(16:0 / 18:2), PI(16:0 / 20:3), PI(16:0 / 20:4), PI(18:0 / 18:1), PI(18:0 / 18:2), PI(18:0 / 20:2), PI(18:0 / 20:3), PI(18:0 / 20:4), PI(18:0 / 22:6), and PI(18:1 / 16:0)) by at least 250% or between 250% and 1000%. In some cases, administration of an anti-EL antibody or antigen-binding fragment thereof may be inducible by administration of at least 30 plasma PI species (e.g., PI(14:2 / 20:0), PI(14:2 / 22:0), PI(14:2 / 22:1), PI(14:2 / 22:2), PI(16:0 / 16:1), PI(16:0 / 18:0), PI(16:0 / 18:2), PI(16:0 / 20:2), PI(16:0 / 20:3), PI(16:0 / 20:4), PI(16:0 / 22:4), PI(16:1 / 18:0), PI(16:1 / 18:1), PI(18:0 / 18:0), PI(18: Increase levels of PI(18:1 / 16:0), PI(18:1 / 18:1), PI(18:0 / 18:2), PI(18:0 / 18:3), PI(18:0 / 20:2), PI(18:0 / 20:3), PI(18:0 / 20:4), PI(18:0 / 22:4), PI(18:0 / 22:5), PI(18:0 / 22:6), PI(18:1 / 16:0), PI(18:1 / 18:1), PI(18:1 / 18:2), PI(18:1 / 20:2), PI(18:1 / 20:3), PI(18:1 / 20:4), and PI(18:2 / 18:2) by at least 250% or between 250% and 1000%.

[0107] Increases in plasma PI may occur within 90 days of the first dose.

[0108] In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 100 mg to about 350 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 100 mg to about 250 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 100 mg to about 200 mg. The dose of the antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, can be administered parenterally, for example, subcutaneously. The dose of the antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof can be administered using an attached pre-filled syringe (APFS) or an autoinjector. The dose of the antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, can be administered about once a month.

[0109] In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 200 mg to about 350 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 200 mg to about 300 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 200 mg to about 250 mg. The dose of the antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, can be administered parenterally, for example, subcutaneously. The dose of the antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof can be administered using an attached pre-filled syringe (APFS) or an autoinjector. The dose of the antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, can be administered about once a month.

[0110] In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 250 mg to about 300 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 250 mg to about 350 mg. The dose of the antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, can be administered parenterally, for example, subcutaneously. The dose of the antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, can be administered using an attached pre-filled syringe (APFS) or an auto-injector. Antibody or an antigen-binding fragment thereof, or a pharmaceutical composition thereof, can be administered about once a month.

[0111] In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 100 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 110 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 120 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 125 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 130 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 140 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 150 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 160 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 170 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 175 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 180 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 190 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 200 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 210 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 220 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 225 mg.In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 230 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 240 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 250 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 260 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 270 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 275 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 280 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 290 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 300 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 310 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 320 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 325 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 330 mg. In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 340 mg. In some embodiments, the anti-EL antibody or The antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 350 mg. The antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, dose can be administered parenterally, for example, subcutaneously. The antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, dose can be administered using an attached pre-filled syringe (APFS) or auto-injector. The antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, dose can be administered about once a month.

[0112] In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 125 mg or 125 mg. The antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof at a dose of about 125 mg or 125 mg, can be administered parenterally, for example, subcutaneously. The antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof at a dose of about 125 mg or 125 mg, can be administered using a pre-filled syringe (APFS) or auto-injector.

[0113] In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 250 mg or 250 mg. The antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof at a dose of about 250 mg or 250 mg, can be administered parenterally, for example, subcutaneously. The antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof at a dose of about 250 mg or 250 mg, can be administered using an attached pre-filled syringe (APFS) or an auto-injector.

[0114] In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered about once a month or once a month. The antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, administered about once a month or once a month, can be administered parenterally, for example, subcutaneously. The antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, administered about once a month or once a month, can be administered using an attached pre-filled syringe (APFS) or an auto-injector.

[0115] In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 125 mg about once per month. The dose of about 125 mg administered about once per month can be administered parenterally, for example, subcutaneously. The dose of about 125 mg administered about once per month can be administered using a pre-filled syringe (APFS) or an auto-injector.

[0116] In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of about 250 mg about once per month. The dose of about 250 mg administered about once per month can be administered parenterally, for example, subcutaneously. The dose of about 250 mg administered about once per month can be administered using a pre-filled syringe (APFS) or an auto-injector.

[0117] In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of 125 mg once a month. The 125 mg dose administered once a month can be administered parenterally, for example, subcutaneously. The approximately 125 mg dose administered approximately once a month can be administered using a pre-filled syringe (APFS) or auto-injector.

[0118] In some embodiments, the anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof, is administered at a dose of 250 mg once per month. The 250 mg dose administered once per month can be administered parenterally, e.g., subcutaneously. About once per month The approximately 250 mg dose can be administered using the provided pre-filled syringe (APFS) or auto-injector.

[0119] According to the methods provided herein, an anti-EL antibody or antigen-binding fragment thereof, or a pharmaceutical composition comprising an anti-EL antibody or antigen-binding fragment thereof, can be administered parenterally. In some aspects of the present disclosure, an anti-EL antibody or antigen-binding fragment thereof, or a pharmaceutical composition comprising an anti-EL antibody or antigen-binding fragment thereof, is administered subcutaneously.

[0120] In some embodiments of the present disclosure, the anti-EL antibody or antigen-binding fragment thereof is administered for at least 3 months (e.g., about once per month). In some embodiments of the present disclosure, the anti-EL antibody or antigen-binding fragment thereof is administered for at least 12 months (e.g., about once per month). In some embodiments of the present disclosure, the anti-EL antibody or antigen-binding fragment thereof is administered for at least 24 months (e.g., about once per month).

[0121] In some aspects of the present disclosure, the present disclosure relates to an anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition provided herein for use as a medicament administered at about 100 mg to about 350 mg or about 200 mg to about 350 mg (e.g., 250 mg). In some aspects of the present disclosure, the present disclosure relates to an anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition provided herein for use as a medicament administered at about 100 mg to about 350 mg or about 200 mg to about 350 mg (e.g., about 250 mg).

[0122] In some aspects of the disclosure, the present disclosure relates to an anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition provided herein for use as a medicament administered at about 100 mg to about 350 mg or about 200 mg to about 350 mg (e.g., 250 mg) approximately once per month. In some aspects of the disclosure, the present disclosure relates to an anti-EL antibody or antigen-binding fragment thereof, or pharmaceutical composition provided herein for use as a medicament administered at about 100 mg to about 350 mg or about 200 mg to about 350 mg (e.g., about 250 mg) approximately once per month.

[0123] According to the methods provided herein, an anti-EL antibody or an antigen-binding fragment thereof, or a pharmaceutical composition containing an anti-EL antibody or an antigen-binding fragment thereof, can be administered in combination with a PCSK9 inhibitor. PCSK9 inhibitors are described, for example, in Chaudhary et al., World J. Cardiol. 9: 76-91 (2017) and Chodorge et al., Sci. Rep. 8: 17545 (2018) (each of which is incorporated herein by reference). The PCSK9 inhibitor can be, for example, an antibody or antigen-binding fragment thereof that binds to PCSK9. Examples of antibodies or antigen-binding fragments thereof that inhibit PCSK9 include HS9, alirocumab, evolocumab, and bococizumab.

[0124] The HS9 antibody (in the context of a GLP-1 fusion protein called MEDI4166) is disclosed in Chodorge et al., Sci. Rep. 8: 17545 (2018) and PCT International Publication No. WO 2015 / 127273, each of which is incorporated herein by reference in its entirety. The HS9 antibody comprises the following variable heavy and light chain sequences: HS9 variable heavy chain sequence: QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGEISPSGGSTSYNQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARERPLYASDLWGQGTTVTVSS (SEQ ID NO: 14) HS9 variable light chain sequence: DIQMTQSPSSLSASVGDRVTITCQASQDVKTAVAWYQQKPGKAPKLLIYSASYRYTGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQRYSLWRTFGQGTKLEIK (SEQ ID NO: 15).

[0125] In some embodiments of the disclosure, the antibody or antigen-binding fragment thereof that inhibits PCSK9 comprises the variable heavy chain sequence of SEQ ID NO: 14. In some embodiments of the disclosure, the antibody or antigen-binding fragment thereof that inhibits PCSK9 comprises the variable light chain sequence of SEQ ID NO: 15. In some embodiments of the disclosure, the antibody or antigen-binding fragment thereof that inhibits PCSK9 comprises the variable heavy chain sequence of SEQ ID NO: 14 and the variable light chain sequence of SEQ ID NO: 15.

[0126] In some embodiments of the present disclosure, the antibody that inhibits PCSK9 comprises a human IgG1 heavy chain. In some embodiments of the present disclosure, the antibody that inhibits PCSK9 comprises a human IgG1 heavy chain comprising the triple mutation L234F / L235E / P331S ("IgG1-TM"). In some embodiments of the present disclosure, the antibody that inhibits PCSK9 comprises a human kappa light chain. In some embodiments of the present disclosure, the antibody that inhibits PCSK9 comprises a) an IgG1-TM heavy chain comprising the variable heavy chain sequence of SEQ ID NO: 14, and b) a kappa light chain comprising the variable light chain sequence of SEQ ID NO: 15.

[0127] In some embodiments of the present disclosure, inhibitors of PCSK9 can promote LDL-C uptake in HepG2 cells treated with recombinant PCSK9 (e.g., as disclosed in Chodorge et al., Sci. Rep. 8: 17545 (2018)).

[0128] As provided herein, the inhibitor of PCSK9 can be administered simultaneously (in the same pharmaceutical composition or in separate pharmaceutical compositions) or sequentially with the anti-EL antibody or antigen-binding fragment thereof.

[0129] 5.3 EL Antibodies and Antigen-Binding Fragments Thereof Provided herein are methods of treating cardiovascular disease in a subject (e.g., a human subject), comprising administering to the subject an antibody (e.g., a monoclonal antibody, such as a chimeric antibody, a humanized antibody, or a human antibody) and antigen-binding fragments thereof that specifically bind to EL (e.g., human EL). Exemplary EL antibodies and antigen-binding fragments thereof that can be used in the methods provided herein are known in the art. The amino acid sequence for human EL is known in the art, and the mature form of the protein (lacking the leader sequence) is provided herein as the sequence of SEQ ID NO: 11.

[0130] Mature human EL (lacking leader sequence): SPVPFGPEGRLEDKLHKPKATQTEVKPSVRFNLRTSKDPEHEGCYLSVGHSQPLEDCSFNMTAKTFFIIHGWTMSGIFENWLHKLVSALHTREKDANVVVVDWLPLAHQLYTDAVNNTRVVG HSIARMLDWLQEKDDFSLGNVHLIGYSLGAHVAGYAGNFVKGTVGRITGLDPAGPMFEGADIHKRLSPDADFVDVLHTYTRSFGLSIGIQMPVGHIDIYPNGGDFQPGCGLNDVLGSIAYG TITEVVKCEHERAVHLFVDSLVNQDKPSFAFQCTDSNRFKKGICLSCRKNRCNSIGYNAKKMRNKRNSKMYLKTRAGMPFRVYHYQMKIHVFSYKNMGEIEPTFYVTLYGTNADSQTLPLEIVERIEQNATNTFLVYTEEDLGDLLKIQLTWEGASQSWYNLWKEFRSYLSQPRNPGRELNIRRIRVKSGETQRKLTFCTEDPENTSISPGRELWFRKCRDGWRMKNETSPTVELP (SEQ ID NO: 11).

[0131] In some embodiments of the present disclosure, the antibodies or antigen-binding fragments thereof used in the methods described herein specifically bind to human EL. In some embodiments of the present disclosure, the antibodies or antigen-binding fragments thereof used in the methods described herein specifically bind to human EL and cynomolgus EL.

[0132] In some embodiments of the present disclosure, the anti-EL antibody or antigen-binding fragment thereof binds to human EL with an equilibrium dissociation constant (KD) of about 4.06 nM (e.g., as measured using surface plasmon resonance). The fragment binds to cynomolgus monkey EL with a KD of about 1.56 nM (e.g., measured using surface plasmon resonance). In some embodiments of the present disclosure, the anti-EL antibody or antigen-binding fragment thereof binds to human EL with a KD constant of about 4.06 nM and binds to cynomolgus monkey EL with a KD of about 1.56 nM (e.g., measured using surface plasmon resonance).

[0133] In some aspects of the present disclosure, the anti-EL antibody or antigen-binding fragment thereof is capable of neutralizing or inhibiting EL.

[0134] The ability of anti-EL antibodies or their antigen-binding fragments to neutralize EL can be determined using the following protocol: Conditioned medium is incubated in a half-area 96-well microplate containing assay buffer (20 mM Tris-HCl, 150 mM NaCl, 4 mM CaCl, 0.5% BSA) and HDL (e.g., human HDL) at concentrations ranging from 1000 nM to 31.6 pM in the presence or absence of anti-EL antibodies or their antigen-binding fragments for 2 hours at 37°C. Following this incubation, free fatty acid release is measured in each well using the NEFA-HR(2) assay kit (Wako Diagnostics, Mountain View, CA). Absorbance at both 550 nm and 660 nm is measured using a SpectraMax M5 plate reader. The value obtained by subtracting the 660 nm reading from the 550 nm reading is used for further analysis. The percent activity "Ax" at antibody concentration "x" is calculated using the following equation: Ax = [(Ex-V0) / (E0-V0)] × 100 (where "Ex" is the average absorbance units in the presence of enzyme with inhibitor, and "E0" and "V0" are the average absorbance units without inhibitor and in the absence of enzyme, respectively. The 50% inhibitory concentration (IC50) is calculated and plotted using GraphPad Prism.

[0135] The same assay can be performed using VLDL (e.g., human VLDL) instead of HDL (e.g., human HDL) as the substrate to show that anti-EL antibodies do not neutralize hepatic lipase or lipoprotein lipase.

[0136] In some embodiments of the present disclosure, the anti-EL antibody or antigen-binding fragment thereof has a half maximal inhibitory concentration (IC) of about 1.3 nM against human EL. 50 In some embodiments of the present disclosure, the anti-EL antibody or antigen-binding fragment thereof has an IC of about 1.7 nM against cynomolgus EL. 50 In some embodiments of the present disclosure, the anti-EL antibody or antigen-binding fragment thereof has an IC of about 1.3 nM against human EL. 50 and an IC of about 1.7 nM against cynomolgus monkey EL 50 It has.

[0137] In some embodiments of the present disclosure, the anti-EL antibody or antigen-binding fragment thereof does not inhibit very low density lipoprotein (VLDL) lipolysis mediated by either hepatic lipase or lipoprotein lipase.

[0138] In some embodiments of the present disclosure, the anti-EL antibody or antigen-binding fragment thereof does not block the exchange of cholesterol from HDL particles to LDL particles.

[0139] In some embodiments of the present disclosure, the anti-EL antibody or antigen-binding fragment thereof increases the delivery of low density lipoprotein (LDL) to the low density lipoprotein receptor (LDLR).

[0140] In some aspects of the present disclosure, the antibodies or The antigen-binding fragment specifically binds to human EL and contains the six CDRs of the MEDI5884 antibody, listed as shown in Tables 1 and 2.

[0141] [Table 1]

[0142] [Table 2]

[0143] In some embodiments of the present disclosure, the antibody or antigen-binding fragment thereof used in the methods described herein specifically binds to human EL and comprises the VH of the MEDI5884 antibody listed in Table 3.

[0144] [Table 3]

[0145] In some embodiments of the present disclosure, the antibody or antigen-binding fragment thereof used in the methods described herein specifically binds to human EL and comprises the VL of the MEDI5884 antibody listed in Table 4.

[0146] [Table 4]

[0147] In some embodiments of the present disclosure, the antibody or antigen-binding fragment thereof used in the methods described herein specifically binds human EL and comprises the VH and VL of the MEDI5884 antibody listed in Tables 3 and 4.

[0148] In some embodiments of the present disclosure, the antibody or antigen-binding fragment thereof used in the methods described herein specifically binds to human EL and comprises the heavy chain sequence of the MEDI5884 antibody listed in Table 5.

[0149] [Table 5]

[0150] In some embodiments of the present disclosure, the antibody or antigen-binding fragment thereof used in the methods described herein specifically binds to human EL and comprises the light chain sequence of the MEDI5884 antibody listed in Table 6.

[0151] [Table 6]

[0152] In some embodiments of the present disclosure, the antibody or antigen-binding fragment used in the methods described herein specifically binds to human EL and comprises the heavy and light chain sequences of the MEDI5884 antibody listed in Tables 5 and 6.

[0153] In some embodiments of the present disclosure, an antibody or antigen-binding fragment thereof used in the methods described herein is described by its VL domain alone or its VH domain alone, or by its three VL CDRs alone or its three VH CDRs alone. For example, see Rader C et al., (1998) PNAS 95: 8910-8915 (cited herein), which describes the humanization of a murine anti-αvβ3 antibody by identifying complementary light or heavy chains from a human light or heavy chain library, respectively, thereby obtaining humanized antibody variants with affinities as high as or higher than those of the original antibody. See Clackson T et al., (1991) Nature 352: 624-628 (herein incorporated by reference in its entirety), which describes a method for generating antibodies that specifically bind to a particular antigen by using a particular VL domain (or VH domain) and screening a library for complementary VH or (VL domain). See also Kim SJ & Hong HJ, (2007) J Microbiol 45: 572-577 (incorporated herein in its entirety), which describes a method for generating antibodies that specifically bind to a particular antigen by using a particular VH domain and screening a library (e.g., a human VL library) for complementary VL domains; the selected VL domains could in turn be used to guide the selection of additional complementary (e.g., human) VH domains.

[0154] In some embodiments, the CDRs of an antibody or antigen-binding fragment thereof can be determined according to the Chothia numbering scheme, which refers to the positions of the structural loops of an immunoglobulin (see, e.g., Chothia C & Lesk AM, (1987), J Mol Biol 196: 901-917; Al-Lazik ani B et al., (1997) J Mol Biol 273: 927-948, Chothia C et al., (1992) J Mol Biol 227: 799-817, Tramontano A et al., (1990) J Mol Biol 215(1): 175-82, and the United States (See U.S. Patent No. 7,709,226.) Typically, using the Kabat numbering convention, the Chothia CDR-H1 loop is located at amino acids 26 to 32, 33, or 34 of the heavy chain, the Chothia CDR-H2 loop is located at amino acids 52 to 56 of the heavy chain, and the Chothia CDR-H3 loop is located at amino acids 95 to 102 of the heavy chain, while the Chothia CDR-L1 loop is located at amino acids 24 to 34 of the light chain, the Chothia CDR-L2 loop is located at amino acids 50 to 56 of the light chain, and the Chothia CDR-L3 loop is located at amino acids 89 to 97 of the light chain. The end of the Chothia CDR-H1 loop, when numbered using the Kabat numbering convention, varies between H32 and H34 depending on the length of the loop (this is because the Kabat numbering scheme places the insertion at H35A and H35B; if neither 35A nor 35B is present, the loop ends at 32; if only 35A is present, the loop ends at 33; if both 35A and 35B are present, the loop ends at 34).

[0155] In some embodiments, provided herein are methods of administering antibodies and antigen-binding fragments thereof that specifically bind to EL (e.g., human EL) and comprise the Chothia VH CDRs and VL CDRs of the MEDI5884 antibody listed in Tables 3 and 4. In some embodiments of the present disclosure, provided herein are methods of administering antibodies or antigen-binding fragments thereof that specifically bind to EL (e.g., human EL) and comprise one or more CDRs, wherein the Chothia CDR and Kabat CDR have the same amino acid sequence. In some embodiments of the present disclosure, provided herein are methods of administering antibodies and antigen-binding fragments thereof that specifically bind to EL (e.g., human EL) and comprise a combination of Kabat CDRs and Chothia CDRs.

[0156] In some embodiments of the present disclosure, the CDRs of an antibody or antigen-binding fragment thereof are determined according to the IMGT numbering system described in Lefranc MP, (1999) The Immunologist 7: 132-136 and Lefranc MP et al., (1999) Nucleic Acids Res 27: 209-212. According to the IMGT numbering scheme, VH-CDR1 is located at positions 26-35, VH-CDR2 is located at positions 51-57, VH-CDR3 is located at positions 93-102, VL-CDR1 is located at positions 27-32, VL-CDR2 is located at positions 50-52, and VL-CDR3 is located at positions 89-97. In some embodiments of the present disclosure, provided herein are methods of administering antibodies and antigen-binding fragments thereof that specifically bind to EL (e.g., human EL) and include the IMGT VH CDRs and VL CDRs of the MEDI5884 antibody listed in Tables 3 and 4, e.g., as described in Lefranc MP (1999), supra, and Lefranc MP (1999), supra.

[0157] In some embodiments, the CDRs of the antibody or antigen-binding fragment thereof are synthesized using the methods described in MacCallum RM et al., (1996) J Mol Biol 262: 732-745. See also, for example, "Protein Sequence and Structure Analysis of Antibody Variable Domains" by Martin A. in Antibody Engineering, Kontermann and Duebel (eds.), Chapter 31, pp. 422-439, Springer-Verlag, Berlin (2001). In some embodiments of the present disclosure, antibodies that specifically bind to EL (e.g., human EL) and are listed in Tables 3 and 4 as determined by the method in MacCallum RM et al. Provided herein are methods of administering antibodies or antigen-binding fragments thereof comprising the VH and VL CDRs of the recited MEDI5884 antibody.

[0158] In some embodiments, the CDRs of the antibody or antigen-binding fragment thereof are selected from the group consisting of CDRs that correspond to intermediates between the Kabat CDRs and the Chothia structural loops and ... The AbM numbering scheme, which refers to the AbM hypervariable regions used by the AbM antibody modeling software (Oxford Molecular Group, Inc.), can be determined according to the AbM numbering scheme. In some aspects of the present disclosure, provided herein are methods of administering an antibody or antigen-binding fragment thereof that specifically binds to EL (e.g., human EL) and comprises the VH and VL CDRs of the MEDI5884 antibody listed in Tables 3 and 4 as determined by the AbM numbering scheme.

[0159] In some aspects of the disclosure, provided herein are methods of administering an antibody comprising a heavy chain and a light chain.

[0160] With respect to the heavy chain, in some embodiments of the present disclosure, the heavy chain is a gamma heavy chain. The constant region of the human IgG4P heavy chain may comprise the following amino acid sequence: ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 12).

[0161] In some embodiments of the present disclosure, an antibody that immunospecifically binds to EL (e.g., human EL) used in the methods described herein comprises a heavy chain whose amino acid sequence of the VH domain comprises the CDR amino acid sequences set forth in Table 1, and wherein the constant region of the heavy chain comprises the amino acid sequence of a human gamma (γ) heavy chain constant region, e.g., human IgG4P.

[0162] In some embodiments of the present disclosure, an antibody that immunospecifically binds to EL (e.g., human EL) used in the methods described herein comprises a heavy chain in which the amino acid sequence of the VH domain comprises the amino acid sequence set forth in Table 3, and wherein the constant region of the heavy chain comprises the amino acid sequence of a human gamma (γ) heavy chain constant region, e.g., human IgG4P.

[0163] With respect to the light chain, in some embodiments of the present disclosure, the light chain of the antibody described herein is a kappa light chain. The constant region of a human C kappa light chain can comprise the following amino acid sequence: RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 13).

[0164] In some embodiments of the present disclosure, an antibody that immunospecifically binds to EL (e.g., human EL) used in the methods described herein comprises a light chain whose amino acid sequence of the VL domain comprises the CDR amino acid sequences set forth in Table 2, and wherein the constant region of the light chain comprises the amino acid sequence of a human Ckappa light chain constant region.

[0165] In some embodiments of the present disclosure, an antibody that immunospecifically binds to EL (e.g., human EL) used in the methods described herein comprises a light chain whose VL domain amino acid sequence comprises a sequence set forth in Table 4, wherein the constant region of the light chain comprises the amino acid sequence of a human Ckappa light chain constant region.

[0166] In some embodiments of the present disclosure, antibodies that immunospecifically bind to EL (e.g., human EL) used in the methods described herein comprise a VH domain and a VL domain comprising the amino acid sequence of any of the VH domains and VL domains described herein, wherein the constant region comprises the amino acid sequence of the constant region of an IgG (e.g., human IgG) immunoglobulin molecule. In some embodiments of the present disclosure, antibodies that immunospecifically bind to EL (e.g., human EL) used in the methods described herein comprise a VH domain and a VL domain comprising the amino acid sequence of any of the VH domains and VL domains described herein. wherein the constant region comprises the amino acid sequence of the constant region of an IgG4Pkappa (e.g., human IgG4Pkappa) immunoglobulin molecule.

[0167] As shown herein, antibodies or antigen-binding fragments thereof that immunospecifically bind to EL (e.g., human EL) used in the methods described herein can have reduced effector function, for example, compared to antibodies or antigen-binding fragments having wild-type IgG1 sequences. Reduced effector function can be the result of, for example, the sequence of the constant region of the antibody or antigen-binding fragment thereof.

[0168] As provided herein, antibodies or antigen-binding fragments thereof that immunospecifically bind to EL (e.g., human EL) used in the methods described herein can lack CDC activity and / or ADCC activity, for example, as a result of the sequence of their constant regions.

[0169] In some embodiments of the present disclosure, the antibodies or antigen-binding fragments thereof described herein that immunospecifically bind to EL (e.g., human EL) comprise a heavy chain and a light chain, wherein (i) the heavy chain comprises a VH domain comprising the amino acid sequences of the VH CDR1, VL CDR2, and VL CDR3 of the MEDI5884 antibody listed in Table 1, and (ii) the light chain comprises the VL CDR1, VH CDR2, and VH CDR3 of the MEDI5884 antibody listed in Table 2. (iii) the heavy chain further comprises a constant heavy chain domain comprising the amino acid sequence of the constant domain of a human IgG4P heavy chain, and (iv) the light chain further comprises a constant light chain domain comprising the amino acid sequence of the constant domain of a human kappa light chain.

[0170] In some embodiments of the present disclosure, the antibodies or antigen-binding fragments thereof described herein that immunospecifically bind to EL (e.g., human EL) comprise a heavy chain and a light chain, wherein (i) the heavy chain comprises a VH domain comprising the amino acid sequence of the VH domain of the MEDI5884 antibody listed in Table 3; (ii) the light chain comprises a VL domain comprising the amino acid sequence of the VL domain of the MEDI5884 antibody listed in Table 4; (iii) the heavy chain further comprises a constant heavy chain domain comprising the amino acid sequence of the constant domain of a human IgG4P heavy chain; and (iv) the light chain further comprises a constant light chain domain comprising the amino acid sequence of the constant domain of a human kappa light chain.

[0171] In certain embodiments, the antigen-binding fragment described herein that immunospecifically binds to EL (e.g., human EL) is selected from the group consisting of Fab, Fab', F(ab')2, and scFv, where the Fab, Fab', F(ab')2, or scFv comprises the heavy chain variable region sequence and the light chain variable region sequence of an anti-EL antibody or antigen-binding fragment thereof described herein. The Fab, Fab', F(ab')2, or scFv can be generated by any technique known to those of skill in the art. In some embodiments of the present disclosure, the Fab, Fab', F(ab')2, or scFv further comprises a moiety that extends the half-life of the antibody in vivo. This moiety is also referred to as a "half-life extending moiety." Any moiety known to those of skill in the art that extends the half-life of the Fab, Fab', F(ab')2, or scFv in vivo can be used. For example, the half-life extending moiety can include an Fc region, a polymer, albumin, or an albumin-binding protein or compound. The polymer may include natural or synthetic, optionally substituted, linear or branched polyalkylenes, polyalkenylenes, polyoxylalkylenes, polysaccharides, polyethylene glycols, polypropylene glycols, polyvinyl alcohols, methoxypolyethylene glycols, lactose, amylose, dextran, glycogen, or derivatives thereof. Substituents may include one or more hydroxy, methyl, or methoxy groups. In some embodiments of the present disclosure, Fab, Fab', F(ab')2, or scFv may be prepared by adding one or more C-terminal amino acids. It can be modified by attaching a half-life extending moiety. In some embodiments of the present disclosure, the half-life extending moiety is polyethylene glycol or human serum albumin. In some embodiments of the present disclosure, the Fab, Fab', F(ab')2, or scFv is fused to an Fc region.

[0172] 5.4 Pharmaceutical Compositions Provided herein are methods for administering compositions containing an anti-EL antibody or antigen-binding fragment thereof having a desired purity in a physiologically acceptable carrier, excipient, or stabilizer (Remington's Pharmaceutical Sciences (1990) Mack Publishing Co., Easton, PA). Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations used (see, e.g., Gennaro, Remington: The Science and Practice of Pharmacy with Facts and Comparisons: Drugfacts Plus, 20th Edition (2003); Ansel et al., Pharmaceutical Dosage Forms and Drug Delivery Systems, 7th Edition, Lippencott Williams and Wilkins (2004); Kibbe et al., Handbook of Pharmaceutical (See Excipients, 3rd ed., Pharmaceutical Press (2000)). The compositions employed can be sterile, which is readily accomplished, for example, by filtration through sterile filtration membranes.

[0173] In some embodiments of the present disclosure, methods are provided for administering (i) an isolated antibody or antigen-binding fragment thereof that specifically binds to human EL, comprising: (a) the sequences of heavy chain variable region (VH) complementarity-determining region (CDR) 1, VH CDR2, VH CDR3, and light chain variable region (VL) CDR1, CDR2, and CDR3, respectively, of SEQ ID NOs: 1 to 6; (b) a variable heavy chain region comprising the amino acid sequence of SEQ ID NO: 7, and / or a variable light chain region comprising the amino acid sequence of SEQ ID NO: 8; or (c) a heavy chain comprising the amino acid sequence of SEQ ID NO: 9, and / or a light chain comprising the amino acid sequence of SEQ ID NO: 10; and (ii) a pharmaceutical composition comprising a pharmaceutically acceptable excipient.

[0174] In some embodiments of the present disclosure, the pharmaceutical composition comprising the isolated antibody or antigen-binding fragment thereof that specifically binds to human EL also comprises an inhibitor of PCSK9. In some embodiments of the present disclosure, the pharmaceutical composition comprising the isolated antibody or antigen-binding fragment thereof that specifically binds to human EL is administered in combination with an inhibitor of PCSK9.

[0175] 5.5 Antibody Production and Polynucleotides Antibodies and antigen-binding fragments thereof that immunospecifically bind to EL (e.g., human EL) can be produced by any method known in the art for the synthesis of antibodies and antigen-binding fragments thereof, for example, by chemical synthesis or recombinant expression techniques. The methods described herein employ, unless otherwise indicated, conventional techniques in molecular biology, microbiology, genetic analysis, recombinant DNA, organic chemistry, biochemistry, PCR, oligonucleotide synthesis and modification, nucleic acid hybridization, and related fields that are within the skill of the art. These techniques are described, for example, in the references cited herein and are fully explained in the literature. See, for example, Sambrook J et al., (2001) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Ausubel FM et al., Current Protocols in Molecular See Biology, John Wiley & Sons (1987 and annually updated), Current Protocols in Immunology, John Wiley & Sons (1987 and annually updated), Gait (ed.) (1984) Oligonucleotide Synthesis: A Practical Approach, IRL Press, Eckstein (ed.) (1991) Oligonucleotides and Analogues: A Practical Approach, IRL Press, Birren B et al., (eds.) (1999) Genome Analysis: A Laboratory Manual, Cold Spring Harbor Laboratory Press.

[0176] In some aspects, provided herein are methods of administering an anti-EL antibody or antigen-binding fragment thereof, or a pharmaceutical composition comprising such an antibody or fragment, wherein the antibody or fragment is produced by recombinant expression of a polynucleotide comprising a nucleotide sequence in a host cell.

[0177] In some embodiments, the anti-EL antibody or antigen-binding fragment administered in accordance with the methods provided herein is encoded by a polynucleotide encoding an anti-EL antibody or antigen-binding fragment thereof, or a domain thereof, that has been optimized, e.g., by codon / RNA optimization, replacement with a heterologous signal sequence, and removal of mRNA instability elements. Methods for generating nucleic acids encoding anti-EL antibodies or antigen-binding fragments thereof, or domains thereof (e.g., heavy chain, light chain, VH domain, or VL domain) that have been optimized for recombinant expression by introducing codon changes (e.g., codon changes that encode the same amino acid due to the degeneracy of the genetic code) and / or by removing inhibitory regions within the mRNA can be performed, e.g., by correspondingly adapting the optimization methods described in U.S. Patent Nos. 5,965,726, 6,174,666, 6,291,664, 6,414,132, and 6,794,498.

[0178] The polynucleotide may be, for example, in the form of RNA or in the form of DNA. DNA includes cDNA, genomic DNA, and synthetic DNA. DNA may be double-stranded or single-stranded. If single-stranded, the DNA may be the coding strand or the non-coding (antisense) strand. In some embodiments of the present disclosure, the polynucleotide is a cDNA or a DNA lacking one or more introns. In some embodiments of the present disclosure, the polynucleotide is a non-naturally occurring polynucleotide. In some embodiments of the present disclosure, the polynucleotide is recombinantly produced. In some embodiments of the present disclosure, the polynucleotide is isolated. In some embodiments of the present disclosure, the polynucleotide is substantially pure. In some embodiments of the present disclosure, the polynucleotide is purified from natural components.

[0179] In some embodiments, a vector (e.g., an expression vector) comprises a nucleotide sequence encoding an anti-EL antibody and its antigen-binding fragment or domain thereof for recombinant expression in a host cell, preferably a mammalian cell. In some embodiments, a cell, e.g., a host cell, comprises such a vector that recombinantly expresses an anti-EL antibody or antigen-binding fragment thereof (e.g., a human antibody or humanized antibody or antigen-binding fragment thereof) described herein. Thus, methods of producing an antibody or antigen-binding fragment thereof described herein can include expressing such an antibody or antigen-binding fragment thereof in a host cell.

[0180] The expression vector can be transferred into cells (e.g., host cells) by conventional techniques, and the resulting cells can then be cultured by conventional techniques to produce an antibody or antigen-binding fragment thereof described herein (e.g., an antibody or antigen-binding fragment thereof comprising the six CDRs, VH, VL, VH and VL, heavy chain, light chain, or heavy and light chain of MEDI5884) or a domain thereof (e.g., VH, VL, VH and VL, heavy chain, or light chain of MEDI5884).

[0181] In some embodiments of the present disclosure, an anti-EL antibody or antigen-binding fragment thereof (e.g., an antibody or antigen-binding fragment thereof comprising the CDRs of MEDI5884) is administered according to the methods provided herein and produced in a host cell. In some embodiments of the present disclosure, the host cell is a CHO cell.

[0182] In some embodiments of the present disclosure, the antibodies or antigen-binding fragments thereof administered in accordance with the methods provided herein are isolated or purified. Generally, an isolated antibody or antigen-binding fragment thereof is substantially free of other antibodies or antigen-binding fragments thereof that have antigenic specificities different from those of the isolated antibody or antigen-binding fragment thereof. For example, in some embodiments of the present disclosure, preparations of antibodies or antigen-binding fragments thereof described herein are substantially free of cellular material and / or chemical precursors.

[0183] The following examples are offered by way of illustration and not by way of limitation. [Example]

[0184] 6. Working Example The examples in this section (ie, Section 6) are offered by way of illustration and not by way of limitation.

[0185] 6.1 Example 1: Nonclinical Pharmacology of MEDI5884 A nonclinical in vivo pharmacology study demonstrated that administration of a single subcutaneous (SC) dose of MEDI5884 (0.5 mg / kg, 6 mg / kg, or 30 mg / kg) in normal male cynomolgus monkeys resulted in a dose-dependent increase in plasma HDL-C. The increases in HDL-C from baseline to maximum effect for the 0.5 mg / kg, 6 mg / kg, and 30 mg / kg doses were 63±14 mg / dL to 96±29 mg / dL, 60±3.8 mg / dL to 111±5.6 mg / dL, and 54±7.4 mg / dL to 122±17 mg / dL, respectively. Smaller increases in total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), and non-HDL-C (TC minus HDL-C) were also observed.

[0186] Apolipoprotein A1 (ApoA1), the major lipoprotein component of HDL, was also increased by 75 ± 5.5% at the 30 mg / kg dose. A dose-dependent increase in serum phospholipids was observed, indicating that MEDI5884 inhibited the hydrolysis of phospholipids contained in HDL particles.

[0187] Increases in ATP-binding cassette transporter A1 (ABCA1), cholesterol efflux capacity (CEC), overall cholesterol efflux, and total number of HDL particles were observed at the highest dose (30 mg / kg). Increases in the mean number of total and large low-density lipoprotein (LDL) particles were also observed. Cholesterol efflux was measured using HDL obtained from animals in the 0.5 mg / kg and 30 mg / kg MEDI5884 treatment groups at days 0, 0.5, 1, 2, 3, 7, and 14 after dosing. ABCA1 efflux was transiently reduced below baseline (days 0.5 and 1) after 30 mg / kg MEDI5884 treatment but increased by 6%, 26%, and 114% above baseline levels on days 3, 7, and 14, respectively. Similarly, overall withdrawals temporarily decreased below baseline (-6%) on day 0.5 and increased to 6%, 5%, 41%, and 53% on days 2, 3, 7, and 14, respectively.

[0188] HDL particles were characterized using NMR spectroscopy in samples obtained from cynomolgus monkeys treated with MEDI5884. Treatment with 30 mg / kg MEDI5884 increased the total number of HDL particles by up to 49 ± 6.3%, and the numbers of both small and large particles increased by up to 182 ± 85% and 104 ± 10%, respectively. Medium HDL particles decreased by up to 48 ± 14% on day 2 and returned to baseline by day 49, suggesting speciation or interconversion of HDL particles as a new equilibrium was established. HDL size increased from baseline to plateau levels for the 0.5 mg / kg, 6 mg / kg, and 30 mg / kg doses from 10.0 ± 0.033 nm to 10.9 ± 0.17 nm, 10 ± 0.23 nm to 10.7 ± 0.12 nm, and 9.9 ± 0.033 nm to 10.7 ± 0.13 nm, respectively.

[0189] These data support the use of MEDI5884 in human patients, for example, for the prevention of secondary cardiovascular events.

[0190] 6.2 Example 2: Nonclinical Pharmacokinetics and Safety of MEDI5884 MEDI5884 was administered to cynomolgus monkeys in a single-dose non-Good Laboratory Practice (GLP) PK / pharmacodynamics (PD) study and two repeat-dose GLP toxicity studies. Single-dose administration of MEDI5884 SC at doses up to 30 mg / kg was well tolerated. No unscheduled deaths, adverse clinical findings, injection site reactions, or adverse effects on body weight were observed. After repeated SC administration of MEDI5884 at doses of 10 mg / kg, 30 mg / kg, or 100 mg / kg (one dose every 2 weeks) for 1 month or 6 months, all animals survived until scheduled sacrifice. No MEDI5884-related changes were observed in clinical observations, ophthalmological evaluations, body weight, behavior, neurophysiology, respiratory rate, injection site irritation scoring, heart rate, electrocardiogram (ECG), or blood pressure. Treatment-related findings were limited to pharmacologically mediated minimal to moderate increases in TC, HDL-C, LDL-C, and phospholipids at all dose levels. Microscopy revealed the presence of minimal to moderate perivascular lymphocytic / mixed leukocyte infiltrates at the SC injection site in a small number of MEDI5884-treated animals; no treatment-related histopathological findings were observed in other tissues. Based on these results, the no-observed-adverse-effect level (NOAEL) was determined to be 100 mg / kg per dose.

[0191] 6.3 Example 3: Phase 1 Clinical Evaluation of MEDI5884 MEDI5884 was evaluated in a phase 1, first-in-human, blinded, placebo-controlled, single-ascending-dose (SDE) study to assess the safety, PK, and PD of subcutaneously (SC) administered MEDI5884 in healthy subjects not receiving statin therapy. Subjects were randomized in a 3:1 ratio to receive 30 mg, 100 mg, 300 mg, or 600 mg of MEDI5884 or placebo. Initial cohorts included six MEDI5884 subjects and two placebo subjects per dose level. These cohorts were then replicated using subjects of Japanese ancestry to provide data supporting the conduct of clinical studies in Japan. A total of 64 subjects were enrolled at a single site in the United States (US). Follow-up periods varied between cohorts, ranging from 28 days post-dose to 90 days post-dose.

[0192] Subjects received a single injection of MEDI5884 (or placebo) for the 30 mg and 100 mg doses, three SC injections (100 mg each) for the 300 mg dose, or six SC injections (100 mg each) for the 600 mg dose.

[0193] subject The median (SD) age of enrolled subjects was 35.9 (9) years, 93.9% were male, 53.1% were Asian, 28.1% were Caucasian, 14.1% were Black or African American, 4.7% reported multiple ethnicities, and 90.6% were not of Hispanic ethnicity.

[0194] Pharmacokinetics Following single SC administration of MEDI5884 at doses of 30 mg, 100 mg, 300 mg, and 600 mg, MEDI5884 exhibited nonlinear PK, likely due to target-mediated drug elimination, with greater than dose-proportional Cmax and AUC observed. PK parameters are summarized in Table 7.

[0195] [Table 7]

[0196] The mean PK profiles observed between the Japanese American and general US populations (Western cohort) were nearly overlapping. Higher exposure was observed in Japanese American subjects at the 300 mg dose, likely due to differences in body weight between the Japanese American and general US populations. A moderate effect of body weight on the PK profile was observed in both Western and Japanese American subjects. The CL / F at the 600 mg dose was 0.378 L / day and 0.254 L / day in the general US population (Western cohort) and Japanese American populations. The data suggest that there are no substantial ethnic differences in the PK of MEDI5884.

[0197] Safety and immunogenicity Safety data were evaluated for 64 subjects, including 48 who received MEDI5884 (four dose cohorts of 12 subjects each [30 mg, 100 mg, 300 mg, or 600 mg]; 24 were of Japanese ancestry) and 16 who received placebo (four dose cohorts of four subjects each; 8 were of Japanese ancestry). There were no related treatment-emergent adverse events (TEAEs), treatment-emergent serious adverse events (TESAEs), or deaths leading to withdrawal from the study. TEAEs occurred in similar rates in subjects receiving MEDI5884 (16 / 48, 33.3%) or placebo (5 / 16, 31.3%).

[0198] Although there were no positive ADA results in subjects in the general US population, positive anti-drug antibody (ADA) responses were detected in 6 of 32 Japanese American subjects, including 1 placebo recipient. No AEs were reported to be associated with ADA. Lower exposures were observed in some ADA-positive Japanese American subjects compared with other Japanese American subjects without ADA. Exposures in these ADA-positive subjects were calculated based on the pooled data. Within the range of exposures of subjects at the same dose within the cohort, no effects on PD or safety were observed. In summary, ADAs were rare, were not associated with adverse events (AEs) or effects on PD, and had no clinically relevant effects on PK.

[0199] Pharmacodynamics Overall baseline lipid levels were similar between placebo and MEDI5884-treated subjects.

[0200] In these healthy subjects not receiving statin therapy, substantial increases in HDL-C were observed after MEDI5884 administration. The mean (SD) percent change from baseline in HDL-C at Day 28 was 4.1% (17.4), 42.0% (26.9), 39.7% (22.5), and 49.8% (17.3) compared with 15.9% (16.8) for placebo in subjects receiving MEDI5884 at 30 mg, 100 mg, 300 mg, or 600 mg, respectively.

[0201] Increases in apoA1 were also observed. Small increases in LDL-C and apoB were observed in the pooled population. These increases did not appear to be dose-related and occurred primarily late after dosing. No clear effect on triglyceride levels was observed.

[0202] 6.4 Example 4: Phase 2a Clinical Evaluation of MEDI5884 MEDI5884 was also evaluated in a Phase 2a, randomized, double-blind, placebo-controlled, parallel-design study to assess the safety, PK, PD, and immunogenicity of MEDI5884 in subjects with stable coronary heart disease (CHD) receiving concomitant high-intensity statin therapy and with triglyceride levels ≤500 mg / dL and LDL-C ≤100 mg / dL.

[0203] A total of 132 subjects received three monthly SC doses of placebo or MEDI5884 at doses of 50 mg, 100 mg, 200 mg, 350 mg, and 500 mg.

[0204] subject The study enrolled primarily Caucasian (90.8%) male (87.0%) subjects, with a median age at enrollment of 67 years. Overall baseline characteristics were similar between placebo and MEDI5584-treated subjects. Subjects who received a total of three doses of study drug were: 22 of 23 (95.7%) in the placebo group, and 18 of 20 (90%), 22 of 24 (91.7%), 20 of 22 (90.9%), 20 of 21 (95.2%), and 21 of 22 (95.5%) in the MEDI5584 50 mg, MEDI5584 100 mg, MEDI5584 200 mg, MEDI5584 350 mg, and MEDI5584 500 mg groups, respectively. One subject in the MEDI5884 50 mg group was included as a treatment completer despite only receiving two doses due to a missed Dose 2 visit.

[0205] Pharmacokinetics An interim PK analysis was performed based on data through Day 111. Mean MEDI5884 concentration-time profiles after three monthly SC doses of MEDI5884 are shown by dose cohort in Figure 1A. PK parameters based on noncompartmental analysis are summarized in Table 8.

[0206] [Table 8]

[0207] MEDI5884 exhibited nonlinear PK, likely due to target-mediated drug elimination; however, at 350 mg and 500 mg MEDI5884 doses, PK in the linear range was observed 30 days after administration. Cmax and AUC were more than dose-proportional. Wide intersubject variability and slight drug accumulation were observed. The mean estimated CL / F at 500 mg was 0.389 L / day.

[0208] Safety and immunogenicity There were no deaths or related TESAEs, and AEs were fairly balanced between the MEDI5884-treated (59 of 109 [54.1%]) and placebo (17 of 23 [73.9%]) groups, were not dependent on MEDI5884 dose, and were representative of events expected to occur in the enrolled population. Self-reported injection site reactions occurred in 15 of 109 (14%) MEDI5884-treated subjects and 3 of 23 (13%) placebo-treated subjects. Investigator-reported injection site reactions occurred in 9 of 109 (8%) MEDI5884-treated subjects and 3 of 23 (13%) placebo-treated subjects. These reactions were mild to moderate in severity. Eight subjects discontinued treatment (one placebo recipient and one MEDI5884 recipient withdrew consent for treatment, and six others withdrew for protocol-mandated laboratory observations [some documented as AEs, four documented as elevated apoB, and two documented as elevated triglycerides]).

[0209] ADAs were rare, low potency, not associated with AEs, and had no effect on PK. ADAs were similar in placebo and MEDI5884 recipients. Therefore, observed ADAs may represent false positives.

[0210] Pharmacodynamic effects Overall baseline lipid levels were similar between placebo and MEDI5884-treated subjects.

[0211] The inhibition of hEL by MEDI5884 was found to be dose-dependent (Figure 1B). In particular, the amount of hEL bound by MEDI5884 in human plasma was measured using an immunoassay platform based on Meso Scale Diagnostics (MSD). Briefly, wells on a 96-well plate were coated with MEDI5884 and incubated overnight at 4°C. The next day, the wells were washed with PBS containing 0.05% Tween 20 wash buffer and blocked with I-Block buffer (Applied Biosystems) for 1 hour at room temperature. The plate was washed. Recombinant EL protein standards (Origene Technologies) and human plasma samples were then added to the corresponding wells and incubated for 1 hour at room temperature. After washing, biotinylated EL detection antibody (Origene Technologies) was added to the corresponding wells and incubated for 1 hour at room temperature. After washing the plate, streptavidin-sulfo-TAG antibody (MSD) was added to the corresponding wells and incubated for 1 hour at room temperature. After washing, the wells were inked with reading buffer (MSD). Plates were read using a MESO Sector S 600 plate reader and data were analyzed using the MSD Discovery Workbench software analysis program.

[0212] In these patients, a dose-dependent increase in HDL-C from baseline was observed in the MEDI5884-treated group. The mean (SD) percent change from baseline in HDL-C at Day 91 (using last observation carried forward (LOCF) imputation) was 2.88% (14.86), 21.82% (30.66), 34.41% (34.89), 43.29% (31.09), and 48.31% (25.63) compared with -3.01% (13.60) for placebo in subjects receiving MEDI5884 at 50 mg, 100 mg, 200 mg, 350 mg, or 500 mg, respectively. Figure 2 and Tables 9A and 9B show the observed (non-LOCF) percent change from baseline in HDL-C over the 90-day period and the observed (non-LOCF) change from baseline in HDL-C at Day 91.

[0213] [Table 9A]

[0214] [Table 9B]

[0215] Dose-dependent increases from baseline in HDL particle number and HDL particle size (Table 10) were also observed in the MEDI5884-treated group versus the placebo group.

[0216] [Table 10] TIFF2025131673000013.tif53170

[0217] Dose-dependent increases from baseline in apoA1 (FIG. 3 and Tables 11A and 11B) and high-density lipoprotein phospholipids (HDL-PL) (FIG. 4 and Table 12) were also observed in the MEDI5884-treated group versus the placebo group. In particular, FIG. 3 and Tables 11A and 11B show the observed (non-LOCF) percent change from baseline in ApoA1 over the 90-day period, and the observed (non-LOCF) change from baseline in ApoA1 at day 91. The mean (SD) percent change from baseline in ApoA1 (using LOCF imputation) at Day 91 was 1.32% (14.81), 15.88% (19.66), 24.82% (21.92), 36.26% (27.36), and 36.85% (18.03) compared with 1.42% (11.20) for placebo in subjects receiving MEDI5884 at 50 mg, 100 mg, 200 mg, 350 mg, or 500 mg, respectively.

[0218] [Table 11A]

[0219] [Table 11B]

[0220] [Table 12]

[0221] A dose-dependent increase from baseline in ABCA1-mediated and overall efflux was also observed in the MEDI5884-treated group versus the placebo group. The effect of MEDI5884 on non-ABCA1 cholesterol efflux is shown in Figure 5.

[0222] Slight increases in triglycerides, LDL-C, and apoB were also observed in the MEDI5884-treated group. In the lower dose groups, no clear dose-dependent relationship was observed in the increases in triglycerides, LDL-C, and apoB in the MEDI5884-treated group. Changes in ApoB were significant only at the 500 mg dose. ApoB levels by dose observed at Day 91 are shown in Table 13.

[0223] [Table 13]

[0224] In three subjects treated with MEDI5884, triglycerides increased to near or above 1000 mg / dL. These subjects had additional risk factors for hypertriglyceridemia. No dose-dependent effect of MEDI5884 on triglyceride levels was observed.

[0225] MEDI5884 at the 200 mg dose produced a mean (median) change from baseline of 15.2 (11.0) mg / dL in HDL-C, 6.1 (5.0) mg / dL in LDL-C (direct), and 2 (-1.0) mg / dL in apoB at day 91 compared with placebo (-1.6 [-3.0] mg / dL for HDL-C, -2.6 [-1.0] mg / dL for LDL-C, and -0.5 [0.0] mg / dL for apoB).

[0226] The estimated parameters of the main PK / PD models are summarized in Table 14.

[0227] [Table 14]

[0228] 6.5 Example 5: Phase 2B Clinical Evaluation of MEDI5884 Following the analysis of the data discussed above, a dose of 250 mg SC monthly MEDI5884 was selected for further evaluation.

[0229] More specifically, a closer examination of the data revealed that administration of five doses of MEDI5884 demonstrated a clear dose-dependent increase in exposure in patients (Figure 1A), which resulted in dose-dependent target engagement (i.e., inhibition of EL levels) (Figure 1B). EL levels at the 200 mg dose were not consistently inhibited over approximately 30 days, whereas EL levels at the 350 mg dose maintained complete inhibition during the 30-day dosing interval. Considering the monthly dosing interval, the optimal dose was between the studied doses of 200 mg and 350 mg. It seemed to be between 0 mg.

[0230] Biomarkers such as HDL-C, ApoA1, and HDL-PL increased in a dose-dependent manner after inhibiting EL activity (Figures 2-4). The time course of the biomarkers indicated that the effective dose should be greater than 200 mg. Safety biomarkers within the pathway, such as LDL-C, ApoB, and TG, increased with a less clear dose-dependence than efficacy biomarkers. Increases in ApoB and TG at the 500 mg dose were identified as a concern, suggesting that a dose less than 500 mg should be selected as the optimal dose.

[0231] The multiple comparison procedure modeling (MCP Mod) method was applied to evaluate the relationship of the area under the effect curve (AUEC) of HDL-C, ApoA1, HDL-PL, LDL, ApoB, and TG to dose during the period from day 60 to day 90. The desired biomarkers of HDL-C, ApoA1, and HDL-PL levels reached a maximum with increasing dose (Figures 2-4), thereby determining the dose achieving 90% of the maximum biomarker levels (ED 90 ) can be estimated. 90 The doses were 205 mg, 270 mg, and 265 mg, respectively (Figure 6). Undesirable biomarkers of LDL, ApoB, and TG did not plateau but showed the following trends: (1) LDL levels continued to increase within the range of 50 mg to 500 mg, (2) ApoB levels were independent of dose except at 500 mg, and (3) TG levels remained constant regardless of the administered dose. Thus, results from the MCP Mod approach confirm that monthly administration of 250 mg is likely to achieve 90% of the maximum efficacy levels of desired biomarkers without causing high levels of undesirable biomarkers.

[0232] A mathematical model was developed to describe the pharmacokinetics (PK) of MEDI5884 and the HDL and ApoA1 biomarker profiles following MEDI5884 administration. The PK model portion used a two-compartment PK model with parallel linear and nonlinear elimination pathways, whereas the PD model portion for biomarker modeling followed a typical indirect response model, with inhibition of each biomarker's elimination pathway resulting in an increase in biomarker levels after administration (Figure 7). While this model does not include HDL-P, changes in HDL-P are addressed indirectly through assessment of HDL-C and ApoA1. An increase in HDL-C without an increase in ApoA1 implies larger HDL particles, but not greater particle numbers.

[0233] When PK data from a wide dose range (50 mg to 500 mg) were analyzed simultaneously, MEDI5884 exhibited nonlinear PK due to target-mediated drug elimination that likely saturated at low doses. Therefore, for the simulated PK profile at 250 mg, which was an interpolation between the PK profile observed after the 200 mg dose and the PK profile observed after the 350 mg dose, it is reasonable to assume that PK is linear at higher doses.

[0234] Observed PK / PD data from a Phase 2a study of MEDI5884 in subjects with CHD receiving high-intensity statin therapy were well characterized by this model. Although large intersubject variability was observed in post-dose PK, HDL, and ApoA1, a clear relationship was observed between MEDI5884 exposure and corresponding increases in HDL-C and ApoA1. The model's estimated 50% inhibitory concentration (IC 50 ) value to calculate the IC 90 were calculated (i.e., 3.03 μg / mL and 3.43 μg / mL for HDL-C and ApoA1, respectively). The estimated median trough concentrations after QM at 250 mg were approximately 3.46 μg / mL based on the following simulations, which is the IC for HDL-C and ApoA1. 90 MEDI5884 at a monthly dose of 250 mg The estimated time courses of MEDI5884, HDL, and ApoA1 after administration are shown in FIG.

[0235] Taken together, these data suggest that a 250 mg monthly dose of MEDI5884 demonstrates: (i) linear PK and target association (EL inhibition) over 30 days post-dose, (ii) median trough levels above the IC90 for maximal HDL-C and ApoA1 increases based on PK / PD modeling, and (iii) minimal undesirable LDL-C and apoB increases based on a multiple comparison procedure modeling approach.

[0236] These data therefore support the evaluation of MEDI5884 in a Phase 2b, randomized, double-blind, placebo-controlled study in adults with a history of myocardial infarction (MI) receiving high-intensity statin therapy. These studies, administered at a dose of 250 mg MEDI5884 or placebo once monthly for 24 months, can demonstrate that MEDI5884 reduces the rates of cardiovascular death, MI, stroke, and coronary revascularization.

[0237] 6.6 Example 6: Inhibition of EL and PCSK9 in Cynomolgus Monkeys A combination pharmacology study was conducted to evaluate the impact of proprotein convertase subtilisin / kexin type 9 (PCSK9) inhibition on lipoprotein metabolism following MEDI5884 treatment. The study protocol is shown in Figure 9. To better mimic a hypothetical patient population already medicated with LDL-C-lowering medication, healthy cynomolgus monkeys were first treated with weekly subcutaneous injections of a PCSK9-neutralizing monoclonal antibody (mAb) (10 mg / kg, n=8) or vehicle for 4 weeks, starting on day 0, to establish a low baseline LDL-C. The PCSK9 monoclonal antibody used was HS9 (comprising the VH and VL sequences of SEQ ID NO: 14 and SEQ ID NO: 15, respectively). The HS9 antibody (in the context of a GLP-1 fusion protein called MEDI4166) is disclosed in Chodorge et al., Sci. Rep. 8: 17545 (2018) and PCT International Publication No. WO 2015 / 127273, each of which is incorporated herein by reference in its entirety.

[0238] Four animals from each group were then administered a subcutaneous dose of MEDI5884 (10 mg / kg, n=4) or vehicle (n=4) on days 28 (vertical dashed lines in Figures 10 and 11) and 42. LDL-C, HDL-C, ApoB, and ApoA1 were measured in plasma samples collected at the indicated time points. Overall and ABCA1 efflux were also assessed.

[0239] An increase in LDL-C was observed after EL neutralization. Compared to vehicle-treated animals, PCSK9 inhibition reduced LDL-C by 75% (Figure 10, left graph) and maintained that level of reduction during the 4-week induction phase, but had no apparent effect on HDL-C (Figure 10, right graph). In vehicle-treated animals during the induction phase, MEDI5884 caused increases in both HDL-C and LDL-C. When added in addition to the PCSK9 inhibitor during a second 4-week period, MEDI5884 maintained its ability to raise HDL-C to a similar extent, but the magnitude of the LDL-C increase was significantly slowed, indicating that LDL particles continued to be taken up by the LDL receptor (Figure 10). The patterns of LDL-C and HDL-C paralleled the changes in ApoB and ApoA1, respectively (Figure 11). The effects on overall efflux and ABCA1 efflux are shown in Figure 12. Notably, the observed increases in withdrawal associated with MEDI5884 administration (including MEDI5884+HS9 administration) generally reflect the observed increases in HDL-C.

[0240] These results provide evidence of cholesterol uptake by the LDL receptor and demonstrate that These results demonstrate that the increase in LDL-C observed with MEDI5884 treatment in patients with CKD can be mitigated by mechanisms that upregulate LDL receptors. These results further demonstrate that MEDI5884 can be administered in combination with PCSK9 inhibitors. This combination therapy can exploit the combined action of two complementary mechanisms that both target different aspects of reverse cholesterol transport.

[0241] 6.7 Example 7: Effects of Increasing Doses of MEDI5884 on Plasma Phosphatidylinositol (PI) Levels in Subjects with Stable Coronary Heart Disease The effect of MEDI5884 on plasma phosphatidylinositol (PI) levels in patients with stable coronary heart disease was quantified using a high-throughput multiplex method combining hydrophilic interaction chromatography (HILIC) separation with negative-mode multiple reaction monitoring (MRM). A total of 31 endogenous PI species were monitored.

[0242] reagent All lipid standards were purchased from Avanti Polar Lipids (Alabaster, AL). Three PI standards were used in the study: PI (12:0 / 13:0) was used as the internal standard (IS), and PI (17:0 / 14:1) and PI (21:0 / 22:6) were used as surrogate analytes. High-performance liquid chromatography (HPLC) grade water was purchased from Honeywell (Charlotte, NC). HPLC-grade isopropanol was used. Inool (IPA), acetonitrile (ACN), and ammonia were all purchased from Sigma-Aldrich (St. Louis, MO). Ammonium acetate and bovine serum albumin (BSA) were purchased from MilliporeSigma (Burlington, MA). PBS was purchased from Lonza BioWhittaker (Morristown, NJ). Special glass-coated 96-well extraction plates and glass vials with PTFE (polytetrafluoroethylene) linings were purchased from Thermo-Fisher (Waltham, MA). Human plasma (pooled and individual) was purchased from BioIVT (Westbury, NY).

[0243] Extraction Procedure for Plasma Samples An internal standard (IS) was spiked into isopropanol (IPA) at 60 nM (final concentration) to create an IS-IPA solution. One pooled lot of human plasma was used as a quality control (QC) and prepared at two levels: low QC (LQC) (8-fold dilution with 40 mg / mL bovine serum albumin (BSA) in PBS) and high QC (HQC) (undiluted plasma). All test samples were diluted 8-fold with 40 mg / mL BSA. Eight-fold dilutions of both the HQC and test samples were prepared using an Agilent Bravo automated liquid handling system (Santa Clara, CA) equipped with a Series III 96LT disposable tip head, an automated liquid handling platform. According to the pre-specified plate map, 20 μL of the LQC, HQC, or test sample was transferred to a separate extraction plate and then precipitated with 180 μL of IS-IPA using the automated device described above. Samples were shaken vigorously (900 rpm to 1200 rpm) on an IKA MTS 2 / 4 digital microtiter shaker (Wilmington, NC) for approximately 10 min.

[0244] The samples were then centrifuged at 2500 g for 5 minutes. Using automated liquid handling again, 20 μL of IS-IPA-extracted plasma was transferred to 140 μL of reconstitution solution (90.25% (v / v) acetonitrile (ACN), 9.75% (v / v) water, 10 mM ammonium acetate). The samples were then shaken at 600 rpm for 5 minutes.

[0245] Hydrophilic interaction chromatography separation Chromatographic separations were performed on a Nexera X2 UHPLC system (Shimadzu, Kyoto, Japan) with an Acquity ultra-high performance liquid chromatography (UPLC) BEH (ethylene bridged hybrid) hydrophilic interaction chromatography (HILIC) column (130 Å, 1.7 μm, 2.1 mm × 100 mm, Waters, Milford, MA). The mobile phases used were mobile phase A (MPA) (5% water, 95% ACN, vol / vol) and mobile phase B (MPB) (50% water, 50% ACN, vol / vol), both containing 10 mM ammonium acetate (pH 8.0–8.5). For each sample or QC, 10 μL of the reconstituted IPA extract was injected onto the column. Separations were performed at 37 °C with a flow rate of 0.5 mL / min. The separation gradient was 5% to 13% MPA over 4 min, followed by a 2-min wash period and a 4-min equilibration period.

[0246] mass spectrometry Mass spectrometric detection was achieved using a 6500+ quadrupole ion trap (QTRAP) mass spectrometer (Sciex, Framingham, MA) operated in negative electrospray ionization (ESI) multiple reaction monitoring (MRM) mode. The MS conditions were adjusted and retention times were defined using synthetic reference standard PI species. Structurally distinctive MRM transitions with the highest signal intensity in negative ESI mode, allowing for the identification of the two acyl chains, were selected during the preparation of the synthetic reference standards (surrogate analytes and IS). Structurally similar MRMs, allowing for the identification of the two acyl chains of each endogenous PI species, were then predicted using LipidView (Sciex, Redwood Shores, CA). These are shown in Table 15. The same source parameters were used for all PI species after adjusting the synthetic reference standard. Details of the acquisition method can be found in Table 16.

[0247] [Table 15]

[0248] [Table 16]

[0249] Data collection, analysis, and reporting After acquisition, samples from each acquisition batch were analyzed using the MultiQuant software. The samples were analyzed according to a specified quantification method (.qmethod). Details of the quantification method, including component and outlier settings, can be found in Table 17. Integration and regression settings were optimized for each individual batch based on the peaks of interest generated. However, the same integration and regression parameters were applied to all samples within the same batch. The peak areas of the internal standard and endogenous PI species were calculated for QCs and unknown samples within the batch. The peak area ratios of endogenous PI species were calculated based on the peak area integration in MultiQuant. The MultiQuant quantification result files (.qsession) were then exported to .txt files for further data analysis using Excel (Microsoft Office 2016) and Spotfire (TIBCO™ Spotfire™ Analyst 7.9.2 HF-011 build version 7.9.2.0.12). To assess the linearity of instrument response and measurement precision for each endogenous PI species, the following were evaluated: HQC / LQC ratio, % difference of HQC / LQC ratio from the nominal value, and %CV for HQC and LQC. Most species had %CV values ​​of 30% or less for HQC and LQC values, and the % difference of HQC / LQC ratio from the nominal value was within 70%-130%.

[0250] [Table 17] TIFF2025131673000022.tif238170

[0251] result A total of 978 plasma samples from subjects with stable coronary heart disease were tested as described above. Samples were analyzed in a total of 15 assays. All 15 assays met the acceptance criteria for all PI species, except for PI (16:0 / 16:0), which had consistently unacceptable variability and HQC / LQC ratios. No assays were considered invalid.

[0252] Clinical samples were also obtained and analyzed from healthy volunteers administered MEDI5884. Some batch-to-batch variability was observed. To correct for batch-to-batch variability and explore the possibility of bridging data between clinical studies on CAD patients and healthy volunteers, the SERRF normalization algorithm (Fan S., et al. Anal Chem Mar 5;91 5:3590-6 (2019)) was evaluated and compared favorably with other normalization methods evaluated for this dataset. The results demonstrated that the MEDI5884-treated CAD method outperformed the conventional method. The results, shown in Figures 13A-13C, 14A-14C, 15, and 16, demonstrate that PI levels were significantly higher across all PI species in healthy subjects compared with MEDI5884-untreated CAD patients. This difference was determined to be statistically significant by a two-tailed unequal variance t-test (p<0.0005 for 12 PI species). In the placebo arms of both clinical studies, PI levels remained relatively stable over a period of several months, with lower levels maintained in CAD patients versus healthy volunteers across the 12 PI species tested.

[0253] The levels of various PI species over time relative to visit day in healthy patients and patients with CAD are shown in Figures 13A-13C and 14A-14C. A comparison of median PI species levels in healthy volunteers and CAD patients on day 21 is shown in Figure 15, and a comparison of median PI species levels across days is shown in Figure 16. As shown in Figures 13A-13C, 14A-14C, 15, and 16, after three monthly subcutaneous (SC) doses of MEDI5884, most plasma PI species increased in a dose-dependent manner relative to placebo. The duration of plasma PI increases appeared to correlate with MEDI5884 exposure. For most PI species, increases in PI levels reached saturation at the 350 mg dose level of MEDI5884, and no further increases in PI levels relative to baseline occurred at the 500 mg dose level of MEDI5884. However, at the 200 mg dose level of MEDI5884, increases in PI approached the saturation levels observed in higher-dose cohorts at day 91. The percent change from baseline for each PI species varied from approximately 1000% for less abundant species to approximately 100%-200% for more abundant species for both CAD and healthy volunteer subjects (Figures 17A-17E, 18, and 19A-19E; see Table 18, which summarizes data obtained across all PI species at the indicated doses and visit days, and Tables 19A-19D, which present data obtained for individual PI species). The mean change across PI species for CAD patients reached a maximum increase of approximately 250%-300% for doses of 200 mg and above.

[0254] [Table 18] TIFF2025131673000024.tif35170

[0255] [Table 19A]

[0256] [Table 19B]

[0257] [Table 19C]

[0258] [Table 19D]

[0259] The present invention is not limited in scope by the specific embodiments described herein. Indeed, various modifications of the invention in addition to those described will become apparent to those skilled in the art from the foregoing detailed description and accompanying drawings. Such modifications are intended to be included within the scope of the appended claims.

[0260] All references (e.g., publications, or patents, or patent applications) cited in this specification are incorporated by reference in their entirety for all purposes to the same extent as if each individual reference (e.g., publication, or patent, or patent application) was specifically and individually indicated to be incorporated by reference in its entirety for all purposes.

[0261] Other embodiments are within the scope of the following claims.

Claims

1. A method of treating cardiovascular disease in a subject, comprising administering to the subject about 100 mg to about 350 mg of an antibody or antigen-binding fragment thereof that specifically binds to human endothelial lipase (EL).

2. A method of reducing atherosclerosis in a subject, comprising administering to the subject about 100 mg to about 350 mg of an antibody or antigen-binding fragment thereof that specifically binds to human EL.

3. 1. A method of treating cardiovascular disease or reducing atherosclerosis in a subject, comprising administering to the subject an antibody or antigen-binding fragment thereof that specifically binds to EL, wherein administering the antibody or antigen-binding fragment thereof comprises: (a) increasing high density lipoprotein cholesterol (HDL-C) in said subject; (b) increasing the number of high density lipoprotein (HDL) particles in said subject; (c) increasing HDL particle size in said subject; (d) increasing HDL phospholipids in said subject; (e) increasing apolipoprotein A1 (ApoA1) in said subject; and / or (f) increasing cholesterol efflux capacity (CEC) in said subject.

4. 4. The method of claim 3, wherein the administration reduces the risk of cardiovascular death, non-fatal myocardial infarction (MI), non-fatal stroke, and / or coronary revascularization in subjects with a history of acute coronary syndrome (ACS).

5. 5. The method of claim 3 or 4, wherein the administration prevents a secondary cardiovascular event in the subject.

6. 6. The method of any one of claims 3 to 5, wherein said administering reduces the risk of a major adverse cardiovascular event (MACE) in the subject.

7. A method for reducing the risk of cardiovascular death, non-fatal myocardial infarction (MI), non-fatal stroke, and / or coronary revascularization in a subject with a history of acute coronary syndrome (ACS), comprising administering to the subject an antibody or antigen-binding fragment thereof that specifically binds to human EL.

8. A method of preventing a secondary cardiovascular event in a subject, comprising administering to said subject an antibody or antigen-binding fragment thereof that specifically binds to human EL.

9. A method of reducing the risk of a serious adverse cardiovascular event (MACE) in a subject, comprising administering to the subject an antibody or antigen-binding fragment thereof that specifically binds to human EL.

10. administering the antibody or antigen-binding fragment thereof (a) increasing high density lipoprotein cholesterol (HDL-C) in said subject; (b) increasing the number of high density lipoprotein (HDL) particles in said subject; (c) increasing HDL particle size in said subject; (d) increasing HDL phospholipids in said subject; (e) increasing ApoA1 in said subject; and / or (f) increasing cholesterol efflux capacity (CEC) in the subject.

11. HDL-C, HDL particle number, HDL particle size, HDL phospholipids, ApoE in subjects A method for increasing A1 and / or CEC, the method comprising administering to the subject an antibody or antigen-binding fragment thereof that specifically binds to EL.

12. The method of any one of claims 3 to 10, comprising administering about 100 mg to about 350 mg of the antibody or antigen-binding fragment thereof.

13. 13. The method of any one of claims 1-12, comprising administering about 100 mg, about 110 mg, about 120 mg, about 125 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 175 mg, about 180 mg, about 190 mg, about 200 mg, about 210 mg, about 220 mg, about 225 mg, about 230 mg, about 240 mg, about 250 mg, about 260 mg, about 270 mg, about 275 mg, about 280 mg, about 290 mg, about 300 mg, about 310 mg, about 320 mg, about 325 mg, about 330 mg, about 340 mg, or about 350 mg of the antibody or antigen-binding fragment thereof.

14. The method of any one of claims 1 to 12, comprising administering about 250 mg of the antibody or antigen-binding fragment thereof.

15. 13. The method of any one of claims 1 to 12, comprising administering 250 mg of the antibody or antigen-binding fragment thereof.

16. The method of any one of claims 1 to 12, comprising administering about 200 mg of the antibody or antigen-binding fragment thereof.

17. 13. The method of any one of claims 1 to 12, comprising administering 200 mg to 250 mg of the antibody or antigen-binding fragment thereof.

18. The method of any one of claims 1 to 12, comprising administering about 125 mg of the antibody or antigen-binding fragment thereof.

19. 13. The method of any one of claims 1 to 12, comprising administering 125 mg of the antibody or antigen-binding fragment thereof.

20. 20. The method of any one of claims 1 to 19, wherein the antibody or antigen-binding fragment thereof is administered once a month.

21. 21. The method of claim 20, wherein the antibody or antigen-binding fragment thereof is administered once a month for at least three months.

22. 22. The method of claim 21, wherein the antibody or antigen-binding fragment thereof is administered once a month for at least 12 months or at least 24 months.

23. The method of any one of claims 1 to 22, wherein the antibody or antigen-binding fragment thereof is administered parenterally.

24. 24. The method of claim 23, wherein the antibody or antigen-binding fragment thereof is administered subcutaneously.

25. 25. The method of any one of claims 1 to 24, wherein the antibody or antigen-binding fragment thereof is administered via an attached pre-filled syringe (APFS) or auto-injector.

26. 26. The method of any one of claims 1 to 25, wherein administration of the antibody or antigen-binding fragment thereof inhibits EL in the subject for 30 days.

27. 27. The method of any one of claims 1 to 26, wherein administration of the antibody or antigen-binding fragment thereof increases HDL-C in the subject by at least 30%.

28. 28. The method of claim 27, wherein administration of the antibody or antigen-binding fragment thereof increases HDL-C in the subject by at least 35%.

29. 29. The method of claim 28, wherein administration of the antibody or antigen-binding fragment thereof increases HDL-C in the subject by at least 40%.

30. 30. The method of any one of claims 26 to 29, wherein administration of the antibody or antigen-binding fragment thereof increases HDL-C in the subject within 30 days of the first administration.

31. 30. The method of any one of claims 26 to 29, wherein administration of the antibody or antigen-binding fragment thereof increases HDL-C in the subject within 90 days of the first administration.

32. 32. The method of any one of claims 1 to 31, wherein administration of the antibody or antigen-binding fragment thereof increases ApoA1 in the subject by at least 30%.

33. 33. The method of claim 32, wherein administration of the antibody or antigen-binding fragment thereof increases ApoA1 in the subject by at least 35%.

34. 34. The method of claim 32 or 33, wherein administration of the antibody or antigen-binding fragment thereof increases ApoA1 in the subject within 30 days of the first administration.

35. 34. The method of claim 32 or 33, wherein administration of the antibody or antigen-binding fragment thereof increases ApoA1 in the subject within 90 days of the first administration.

36. 36. The method of any one of claims 1 to 35, wherein administration of the antibody or antigen-binding fragment thereof increases non-ATP-binding cassette transporter A1 (ABCA1) cholesterol efflux capacity in the subject by at least 30%.

37. 37. The method of claim 36, wherein administration of the antibody or antigen-binding fragment thereof increases non-ABCA1 cholesterol efflux capacity in the subject by at least 35%.

38. 38. The method of claim 36 or 37, wherein administration of the antibody or antigen-binding fragment thereof increases non-ABCA1 cholesterol efflux capacity in the subject within 30 days of the first administration.

39. 38. The method of claim 36 or 37, wherein administration of the antibody or antigen-binding fragment thereof increases non-ABCA1 cholesterol efflux capacity in the subject within 90 days of the first administration.

40. 40. The method of any one of claims 1 to 39, wherein administration of the antibody or antigen-binding fragment thereof increases HDL particle number in the subject by at least 5%.

41. 41. The method of claim 40, wherein administration of the antibody or antigen-binding fragment thereof increases HDL particle number in the subject by at least 8%.

42. 42. The method of claim 40 or 41, wherein administration of the antibody or antigen-binding fragment thereof increases HDL particle numbers in the subject within 30 days of the first administration.

43. 42. The method of claim 40 or 41, wherein administration of the antibody or antigen-binding fragment thereof increases HDL particle numbers in the subject within 90 days of the first administration.

44. 44. The method of any one of claims 1 to 43, wherein administration of the antibody or antigen-binding fragment thereof increases HDL particle size in the subject by at least 3%.

45. 45. The method of claim 44, wherein administration of the antibody or antigen-binding fragment thereof increases HDL particle size in the subject by at least 5%.

46. 46. ​​The method of claim 44 or 45, wherein administration of the antibody or antigen-binding fragment thereof increases HDL particle size in the subject within 30 days of the first administration.

47. 46. ​​The method of claim 44 or 45, wherein administration of the antibody or antigen-binding fragment thereof increases HDL particle size in the subject within 90 days of the first administration.

48. 48. The method of any one of claims 1 to 47, wherein administration of the antibody or antigen-binding fragment thereof increases HDL phospholipids in the subject by at least 50%.

49. 48. The method of claim 47, wherein administration of the antibody or antigen-binding fragment thereof increases HDL phospholipids in the subject within 30 days of the first administration.

50. 48. The method of claim 47, wherein administration of the antibody or antigen-binding fragment thereof increases HDL phospholipids in the subject within 90 days of the first administration.

51. 51. The method of any one of claims 1 to 50, wherein administration of the antibody or antigen-binding fragment thereof increases plasma phosphatidylinositol (PI) levels in the subject by at least 100% or at least 250%.

52. The increase in plasma PI levels can be PI (14:2 / 20:0) levels, PI (14:2 / 22:0) levels, PI (14:2 / 22:1) levels, PI (14:2 / 22:2) levels, PI (16:0 / 16:1) levels, PI (16:0 / 18:0) levels, PI (16:0 / 18:2) levels, PI (16:0 / 20:2) levels, PI (16:0 / 20:3) levels, PI (16:0 / 20:4) levels, PI (16:0 / 22:4) levels, PI (16:1 / 18:0) levels, PI (16:1 / 18:1) levels, PI (18:0 / 18:0) levels, PI (18:0 / 18:1) levels, PI (1 52. The method of claim 51 , wherein the PI(18:0 / 18:2) level, PI(18:0 / 18:3) level, PI(18:0 / 20:2) level, PI(18:0 / 20:3) level, PI(18:0 / 20:4) level, PI(18:0 / 22:4) level, PI(18:0 / 22:5) level, PI(18:0 / 22:6) level, PI(18:1 / 16:0) level, PI(18:1 / 18:1) level, PI(18:1 / 18:2) level, PI(18:1 / 20:2) level, PI(18:1 / 20:3) level, PI(18:1 / 20:4) level, and / or PI(18:2 / 18:2) level.

53. 53. The method of claim 51 or 52, wherein administration of the antibody or antigen-binding fragment thereof increases plasma phosphatidylinositol (PI) levels in the subject within 90 days of the first administration.

54. 54. The method of any one of claims 2 to 53, wherein the subject suffers from cardiovascular disease.

55. 55. The method of claim 1 or 54, wherein the cardiovascular disease is coronary artery disease, coronary heart disease, chronic arterial disease, cerebrovascular disease, atherosclerotic cardiovascular disease, or peripheral arterial disease.

56. 54. The method of any one of claims 1 to 53, wherein the subject has stable coronary artery disease or stable coronary heart disease.

57. 54. The method of any one of claims 1 to 53, wherein the subject has a history of acute coronary syndrome (ACS).

58. 58. The method of any one of claims 1 to 57, wherein the subject is undergoing statin therapy.

59. 58. The method of any one of claims 1 to 57, wherein the subject is naive to statin therapy.

60. 60. The method of any one of claims 1-59, wherein the subject has a triglyceride level of 500 mg / dL or less prior to said administering.

61. 61. The method of any one of claims 1-60, wherein the subject has an LDL-C of 100 mg / dL or less prior to said administering.

62. The method of any one of claims 1 to 61, wherein the subject is a human.

63. The method of any one of claims 1 to 62, wherein the antibody or antigen-binding fragment thereof neutralizes EL activity.

64. 64. The method of any one of claims 1 to 63, wherein the antibody or antigen-binding fragment thereof has reduced effector function.

65. 65. The method of any one of claims 1 to 64, wherein the antibody or antigen-binding fragment thereof does not have antibody-dependent cell-mediated cytotoxicity (ADCC) activity.

66. The method of any one of claims 1 to 65, wherein the antibody does not have complement dependent cytotoxicity (CDC) activity.

67. The method of any one of claims 1 to 66, wherein the antibody binds to cynomolgus monkey EL.

68. The method of any one of claims 1 to 67, wherein the antibody or antigen-binding fragment thereof competitively inhibits the binding to EL of an antibody comprising a VH comprising the amino acid sequence set forth in SEQ ID NO: 7 and a VL comprising the amino acid sequence set forth in SEQ ID NO:

8.

69. The method of any one of claims 1 to 68, wherein the antibody or antigen-binding fragment thereof binds to the same epitope of EL as an antibody comprising a VH comprising the amino acid sequence set forth in SEQ ID NO: 7 and a VL comprising the amino acid sequence set forth in SEQ ID NO:

8.

70. 70. The method of any one of claims 1 to 69, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) CDR1, VH CDR2, VH CDR3, a light chain variable region (VL) CDR1, VL CDR2, and VL CDR3 of the sequence of MEDI5884.

71. 71. The method of claim 70, wherein the CDR is a Kabat CDR, a Chothia CDR, or an AbM CDR.

72. 70. The method of any one of claims 1 to 69, wherein the antibody or antigen-binding fragment thereof comprises a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 3, a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a VL CDR3 comprising the amino acid sequence of SEQ ID NO:

6.

73. The method of any one of claims 1 to 72, wherein the antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 7, and / or a VL comprising the amino acid sequence set forth in SEQ ID NO:

8.

74. 77. The method of any one of claims 1 to 76, wherein the antibody or antigen-binding fragment comprises an IgG heavy chain constant region.

75. 75. The method of claim 74, wherein the IgG heavy chain constant region is an IgG4 heavy chain constant region.

76. 76. The method of claim 75, wherein the IgG4 heavy chain constant region is an IgG4P heavy chain constant region.

77. 77. The method of any one of claims 1 to 76, wherein the antibody or antigen-binding fragment thereof comprises a kappa light chain constant region.

78. 77. The method of any one of claims 1 to 76, wherein the antibody or antigen-binding fragment comprises a heavy chain constant region and / or a light chain constant region.

79. 79. The method of claim 78, wherein the heavy chain constant region is a human IgG4P heavy chain constant region and / or the light chain constant region is a human IgGκ light chain constant region.

80. 80. The method of any one of claims 1 to 79, wherein the antibody or antigen-binding fragment thereof is a humanized antibody or antigen-binding fragment thereof.

81. 81. The method of any one of claims 1 to 80, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain constant region comprising the amino acid sequence set forth in SEQ ID NO: 12 and / or a light chain constant region comprising the amino acid sequence set forth in SEQ ID NO:

13.

82. 82. The method of any one of claims 1 to 81, wherein the antibody comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 9 and a light chain comprising the amino acid sequence set forth in SEQ ID NO:

10.

83. 83. The method of any one of claims 1 to 82, wherein the antibody or antigen-binding fragment thereof is a full-length antibody.

84. 81. The method of any one of claims 1 to 80, wherein the antibody or antigen-binding fragment thereof is an antigen-binding fragment.

85. The antigen-binding fragments include Fab, Fab', F(ab') ２ , single-chain Fv (scFv), disulfide-linked Fv, V-NAR domain, IgNar, intrabody, IgGΔCH2, minibody, F(ab') ３ , tetrabodies, triabodies, diabodies, single domain antibodies, DVD-Ig, Fcab, mAb ２ , (scFv) ２ 85. The method of claim 84, comprising:

86. A method for treating cardiovascular disease in a subject, comprising subcutaneously administering 250 mg of an antibody or antigen-binding fragment thereof to the subject once a month, wherein the antibody or antigen-binding fragment specifically binds to human EL and comprises a VH comprising the amino acid sequence set forth in SEQ ID NO:7 and a VL comprising the amino acid sequence set forth in SEQ ID NO:

8.

87. A method for reducing the risk of cardiovascular death, non-fatal myocardial infarction (MI), non-fatal stroke, and / or coronary revascularization in a subject with a history of acute coronary syndrome (ACS), comprising subcutaneously administering 250 mg of an antibody or antigen-binding fragment thereof to the subject once a month, wherein the antibody or antigen-binding fragment specifically binds to human EL and comprises a VH comprising the amino acid sequence set forth in SEQ ID NO:7 and a VL comprising the amino acid sequence set forth in SEQ ID NO:

8.

88. A method for treating cardiovascular disease in a subject, comprising subcutaneously administering approximately 200 mg of an antibody or antigen-binding fragment thereof to the subject once a month, wherein the antibody or antigen-binding fragment specifically binds to human EL and comprises a VH comprising the amino acid sequence set forth in SEQ ID NO:7 and a VL comprising the amino acid sequence set forth in SEQ ID NO:

8.

89. A method for reducing the risk of cardiovascular death, non-fatal myocardial infarction (MI), non-fatal stroke, and / or coronary revascularization in a subject with a history of acute coronary syndrome (ACS), comprising subcutaneously administering approximately 200 mg of an antibody or antigen-binding fragment thereof to the subject once a month, wherein the antibody or antigen-binding fragment specifically binds to human EL and comprises a VH comprising the amino acid sequence set forth in SEQ ID NO:7 and a VL comprising the amino acid sequence set forth in SEQ ID NO:

8.

90. A method for treating cardiovascular disease in a subject, comprising subcutaneously administering to the subject 200 mg to 250 mg of an antibody or antigen-binding fragment thereof once a month, wherein the antibody or antigen-binding fragment specifically binds to human EL and comprises a VH comprising the amino acid sequence set forth in SEQ ID NO:7 and a VL comprising the amino acid sequence set forth in SEQ ID NO:

8.

91. A method for reducing the risk of cardiovascular death, non-fatal myocardial infarction (MI), non-fatal stroke, and / or coronary revascularization in a subject with a history of acute coronary syndrome (ACS), comprising subcutaneously administering to the subject 200 mg to 250 mg of an antibody or antigen-binding fragment thereof once per month, wherein the antibody or antigen-binding fragment specifically binds to human EL and comprises a VH comprising the amino acid sequence set forth in SEQ ID NO:7 and a VL comprising the amino acid sequence set forth in SEQ ID NO:

8.

92. The method of any one of claims 86 to 91, wherein the antibody comprises the amino acid sequence of a heavy chain constant region shown in SEQ ID NO: 9 and the amino acid sequence of a light chain constant region shown in SEQ ID NO:

10.

93. 93. The method of any one of claims 1 to 92, further comprising administering an inhibitor of PCSK9.

94. 94. The method of claim 93, wherein the administration of the antibody or antigen-binding fragment thereof that specifically binds to human EL and the administration of the inhibitor of PCSK9 are simultaneous.

95. 95. The method of claim 94, wherein the antibody or antigen-binding fragment thereof that specifically binds to human EL and the inhibitor of PCSK9 are administered in separate pharmaceutical compositions.

96. 94. The method of claim 93, wherein the administration of the antibody or antigen-binding fragment thereof that specifically binds to human EL and the administration of the inhibitor of PCSK9 are sequential.

97. 97. The method of any one of claims 93 to 96, wherein the inhibitor of PCSK9 is an anti-PCSK9 antibody or an antigen-binding fragment thereof.

98. 98. The method of claim 97, wherein the inhibitor of PCSK9 is HS9, evolocumab, alirocumab, or bococizumab.

Images