Angiopoietin-like protein 4 antibody-based treatment

Abstract

Methods, kits, and compositions are provided herein that can be used to treat diseases, disorders, or conditions using an antibody or antigen-binding fragment thereof that binds specifically to human ANGPTL4.

Description

ANGIOPOIETIN-LIKE PROTEIN 4 ANTIBODY-BASED TREATMENTRELATED APPLICATIONS

[0001] This application claims priority to and the benefit of U.S. Provisional Patent Application No. 63 / 661,011, filed on June 17, 2024; U.S. Provisional Patent Application No. 63 / 744,715, filed on January 13, 2025; U.S. Provisional Patent Application No. 63 / 768,015, filed on March 6, 2025; U.S. Provisional Patent Application No. 63 / 775,940, filed on March 21, 2025; and U.S. Provisional Patent Application No. 63 / 801,047, filed on May 6, 2025, the entire contents of each are hereby incorporated by reference in their entirety.BACKGROUND

[0002] Angiopoietin-like 4 protein (ANGPTL4) is a member of the angiopoietin like family of secreted proteins. It is a homo-oligomeric protein, capable of forming dimers and tetramers that is expressed by cell types including macrophages, adipose, muscle, and liver cells. Lipoprotein lipase (LPL) has a central role in lipoprotein metabolism which includes the maintenance of lipoprotein levels in blood. The coiled-coil region of ANGPTL4 is known to inhibit lipoprotein lipase (LPL)-mediated triglyceride (TG) clearance.

[0003] Elevated plasma triglyceride levels are common and -27% of adults have a fasting plasma triglyceride level >176 mg / dl. Elevated plasma triglyceride levels are associated with an increased risk of myocardial infarction and other adverse cardiovascular events independently of plasma lipoprotein lipase (LDL) cholesterol levels.

[0004] Human carriers of an ANGPTL4 loss-of-function variant (E40K) have lower plasma triglyceride levels and lower atherosclerotic cardiovascular disease (ASCVD) risk, suggesting that ANGPTL4 inhibition could lower triglycerides and ASCVD risk in people with hypertriglyceridemia. Currently, ASCVD-related conditions remain a leading cause of morbidity and mortality globally.

[0005] Accordingly, a need exists for therapeutics that can target ANGPTL4.SUMMARY

[0006] The present disclosure relates, in part, to antibodies or antigen-binding fragments thereof that bind specifically to human angiopoietin-like 4 protein (ANGPTL4), and compositions (such as pharmaceutical compositions) and methods of treatment comprising the same.

[0007] The present disclosure encompasses, the discovery of ANGPTL4 binding antibodies or antigen-binding fragments thereof described herein with novel activity that result in: (i) inhibition of ANGPTL4 binding to lipoprotein lipase (LPL); (ii) lowered circulating triglycerides in human subjects with hypertriglyceridemia; (iii) lowered circulating remnant cholesterol in human subjects with hypertriglyceridemia and / or (iv) lowered circulating fasting insulin levels in human subjects, particularly at the dosage regimens described herein. In some embodiments, an ANGPTL4 binding antibody or antigen-binding fragment thereof described herein blocks ANGPTL4 binding to LPL.

[0008] In a first aspect, disclosed herein is a method of treating a human having a disorder, comprising: subcutaneously administering a dose of about 15 mg to about 1000 mg of an antibody or antigen-binding fragment thereof that binds specifically to human ANGPTL4 about weekly to about annually (e.g., about weekly, about biweekly, or about monthly) to the human, thereby improving at least one sign or symptom of the disorder in the human after administration.

[0009] In some embodiments, a dose is about 15 mg to about 500 mg. In some embodiments, a dose is about 150 mg to about 450 mg. In some embodiments, a dose is about 225 mg to about 450 mg. In some embodiments, a dose is about 150 mg, about 225 mg, about 300 mg, about 450 mg, or about 900 mg. In some embodiments, a dose is about 225 mg or about 450 mg. In some embodiments, a dose is about 225 mg. In some embodiments, a dose is about 450 mg.

[0010] In some embodiments, a dose is administered about weekly to about annually, e.g., about biweekly to about quarterly, about biweekly to monthly or about monthly to about quarterly. In some embodiments, a dose is administered about weekly, about biweekly, about monthly, about quarterly, about biannually, or about annually. In some embodiments, a dose is administered about every 2 weeks to about every 12 weeks, e.g., about every 2 weeks to about every 8 weeks, about every 2 weeks to about every 4weeks, or about every 4 weeks to about every 8 weeks. In some embodiments, a dose is administered about every 2 weeks, about every 3 weeks, about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, or about every 12 weeks. In some embodiments, a dose is administered about every 4 weeks or about every 8 weeks. In some embodiments, a dose is administered about every 4 weeks. In some embodiments, a dose is administered about every 8 weeks.

[0011] In some embodiments, a human is afflicted with a cardiovascular disease. In some embodiments, a cardiovascular disease is or comprises atherosclerotic cardiovascular disease (ASCVD), a myocardial infarction (MI), stroke, coronary revascularization, arteriosclerosis, or any combination thereof.

[0012] In some embodiments, a cardiovascular disease is or comprises ASCVD.

[0013] In some embodiments, a disorder is obesity, fatty liver disease (e.g., nonalcoholic fatty liver disease), insulin resistance, metabolic dysfunction, metabolic syndrome, polycystic ovarian syndrome, chronic kidney disease, Cushing syndrome, hypercorti soli sm, acromegaly, heart failure (e.g., heart failure with preserved ejection fraction), lipodystrophy, diabetes (e.g., type 2 diabetes), or any combination thereof.

[0014] In some embodiments, one or more of fasting plasma triglyceride levels, fasting plasma remnant cholesterol levels, fasting plasma HbAlc levels, fasting plasma insulin levels, and fasting plasma glucose levels are measured prior to administration of one or more doses.

[0015] In some embodiments, one or more of fasting plasma triglyceride levels, fasting plasma remnant cholesterol levels, fasting plasma HbAlc levels, fasting plasma insulin levels, and fasting plasma glucose levels are measured at least about 1 day after administering one or more doses.

[0016] In some embodiments, a step of administering an antibody or antigen-binding fragment thereof described herein results in a reduction in one or more of plasma triglyceride levels, plasma remnant cholesterol levels, plasma HbAlc levels, plasma insulin levels, and plasma glucose levels compared to basal levels.

[0017] In some embodiments, a human is afflicted with lipodystrophy, e.g., familial lipodystrophy or acquired lipodystrophy.

[0018] In some embodiments, a human is afflicted with one or more of hyperlipidemia, hypercholesterolemia, hyperlipoproteinemia, hypertriglyceridemia and dyslipidemia.

[0019] In some embodiments, a human is afflicted with an elevated fasting plasma triglyceride concentration of at least about 150 mg / dl, such as at least about 160 mg / dl, at least about 170 mg / dl, at least about 190 mg / dl, at least about 190 mg / dl, at least about 200 mg / dL, at least about 300 mg / dL, at least about 400 mg / dL, or at least about 500 mg / dL.

[0020] In some embodiments, a human is afflicted with an elevated fasting plasma insulin concentration of at least about 6 plU / ml, such as at least about 7 plU / ml, at least about 8 plU / ml, at least about 9 plU / ml, or at least about 10 plU / ml.

[0021] In some embodiments, an antibody or antigen-binding fragment thereof is administered in combination with a second therapy.

[0022] In some embodiments, a second therapy is an agent described herein.

[0023] In some embodiments, an antibody or antigen-binding fragment thereof is administered to the human in a first cycle followed by a second cycle, wherein the dose administered in the second cycle differs from the dose administered in the first cycle.

[0024] In some embodiments, a first cycle is about 2 weeks to about 6 weeks in duration.

[0025] In some embodiments, a second cycle is about 2 weeks to about 6 weeks in duration.

[0026] In some embodiments, fasting plasma triglyceride levels are measured.

[0027] In some embodiments, fasting remnant cholesterol levels are measured.

[0028] In some embodiments, a reduction in plasma triglyceride levels during and / or after the first cycle compared to basal plasma triglyceride levels results in administering a lower dose in the second cycle compared to the dose administered in the first cycle.

[0029] In some embodiments, comparable or increased plasma triglyceride levels during and / or after the first cycle compared to basal plasma triglyceride level results in administering a higher dose in the second cycle compared to the dose administered in the first cycle.

[0030] In some embodiments, a reduction in plasma remnant cholesterol levels during and / or after the first cycle compared to basal plasma remnant cholesterol levelsresults in administering a lower dose in the second cycle compared to the dose administered in the first cycle.

[0031] In some embodiments, comparable or increased plasma remnant cholesterol levels during and / or after the first cycle compared to basal plasma remnant cholesterol level results in administering a higher dose in the second cycle compared to the dose administered in the first cycle.

[0032] In some embodiments, plasma HbAlc levels are measured.

[0033] In some embodiments, fasting plasma glucose levels are measured.

[0034] In some embodiments, the first cycle comprises administering a dose of about 250 mg to about 500 mg and / or the second cycle comprises administering a dose of about 50 mg to about 250 mg.

[0035] In some embodiments, fasting plasma triglycerides and / or remnant cholesterol are measured at least about 1 day after administering one or more doses, such as at least about 2 days, at least about 3 days, or at least about 4 days after administering one or more doses. In some embodiments, fasting plasma triglycerides and / or remnant cholesterol are measured about 2 days to about 15 days after administering one or more doses.

[0036] In some embodiments, a step of administering an antibody or antigen-binding fragment thereof described herein results in a reduction of fasting plasma triglyceride levels of at least about 10%, such as at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, or at least about 45%, compared to basal fasting plasma triglyceride levels.

[0037] In some embodiments, a step of administering an antibody or antigen-binding fragment thereof described herein results in a reduction of postprandial plasma triglyceride levels of at least about 10%, such as at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, or at least about 45%, compared to basal postprandial plasma triglyceride levels.

[0038] In some embodiments, a step of administering an antibody or antigen-binding fragment thereof described herein results in a reduction of fasting plasma remnant cholesterol levels of at least 10%, such as at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, or at least about 45%, compared to basal fasting plasma remnant cholesterol levels.

[0039] In some embodiments, a step of administering an antibody or antigen-binding fragment thereof described herein results in a reduction of postprandial plasma remnant cholesterol levels of at least about 10%, such as at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, or at least about 45%, compared to basal postprandial plasma remnant cholesterol levels.

[0040] In some embodiments, an antibody or antigen-binding fragment thereof blocks LPL binding to ANGPTL4 protein.

[0041] In some embodiments, an antibody or antigen-binding fragment thereof comprises: (a) a heavy chain variable region (VH) comprising a HCDR1 of SEQ ID NO: 7, a HCDR2 of SEQ ID NO: 8, and a HCDR3 of SEQ ID NO: 9, and a light chain variable region (VL) comprising a LCDR1 of SEQ ID NO: 17, a LCDR2 of SEQ ID NO: 18, and a LCDR3 of SEQ ID NO: 19; or (b) a VH comprising a HCDR1 of SEQ ID NO: 10, a HCDR2 of SEQ ID NO: 11, and a HCDR3 of SEQ ID NO: 12, and a VL comprising a LCDR1 of SEQ ID NO: 20, a LCDR2 of SEQ ID NO: 21, and a LCDR3 of SEQ ID NO: 22.

[0042] In some embodiments, an antibody or antigen-binding fragment thereof is a monoclonal antibody, humanized antibody, single chain antibody, Fab fragment, Fv fragment, F(ab')2 fragment, or scFv fragment. In some embodiments, an antibody or antigen-binding fragment is an IgGl or IgG4 isotype.

[0043] In some embodiments, an antibody or antigen-binding fragment comprises: (i) a VH comprising an amino acid sequence that is at least 90% identical to SEQ ID NO: 13, and (ii) a VL comprising an amino acid sequence that is at least 90% identical to SEQ ID NO: 23. In some embodiments, an antibody or antigen-binding fragment comprises: (i) a VH comprising an amino acid sequence of SEQ ID NO: 13, and (i) a VL comprising an amino acid sequence of SEQ ID NO: 23.

[0044] In some embodiments, an antibody comprises: (i) a heavy chain comprising an amino acid sequence that is at least 90% identical to SEQ ID NO: 28, SEQ ID NO: 156, SEQ ID NO: 157, or SEQ ID NO: 15, and (ii) a light chain comprising an amino acid sequence that is at least 90% identical to SEQ ID NO: 25. In some embodiments, an antibody comprises: (i) a heavy chain comprising an amino acid sequence of SEQ ID NO: 28, SEQ ID NO: 156, SEQ ID NO: 157, or SEQ ID NO: 15, and (ii) a light chain comprising an amino acid sequence of SEQ ID NO: 25.

[0045] In some embodiments, an antibody comprises: (i) a heavy chain comprising an amino acid sequence of SEQ ID NO: 28, and (ii) a light chain comprising an amino acid sequence of SEQ ID NO: 25. In some embodiments, an antibody comprises: (i) a heavy chain comprising an amino acid sequence of SEQ ID NO: 15, and (ii) a light chain comprising an amino acid sequence of SEQ ID NO: 25. In some embodiments, an antibody comprises: (i) a heavy chain comprising an amino acid sequence of SEQ ID NO: 156, and (ii) a light chain comprising an amino acid sequence of SEQ ID NO: 25. In some embodiments, an antibody comprises: (i) a heavy chain comprising an amino acid sequence of SEQ ID NO: 157, and (ii) a light chain comprising an amino acid sequence of SEQ ID NO: 25.

[0046] In some embodiments, an antibody or antigen-binding fragment thereof disclosed herein is formulated in a pharmaceutical composition comprising a pharmaceutically acceptable carrier.BRIEF DESCRIPTION OF THE DRAWING

[0047] The Figures described below, which together make up the Drawing, are for illustration purposes only, not for limitation.

[0048] Figure 1 depicts a clinical human study design. This was a non-confirmatory, first-in-human, randomized, subject-blinded, placebo-controlled, single ascending dose study of LTW980 in healthy subjects. In all parts of the study, subjects were administered a single dose of LTW980 or placebo via subcutaneous (s.c.) injection. Part IA consisted of 4 dose escalation cohorts (15-450 mg LTW980 or placebo, 8 subjects per cohort) in subjects with BMI 18-30 kg / m2. In Part IB, 450 mg LTW980 or placebo was administered to 12 subjects with high BMI in the range of 30-40 kg / m2who were otherwise healthy. In Part IC, 450 mg LTW980 or placebo was administered to 12 subjects weighing at least 59 kg and with triglycerides in the range of 200-500 mg / dL at the screening visit, who were otherwise healthy.

[0049] Figure 2 depicts (A) LTW980 administration lowered fasting plasma triglyceride levels in human subjects with moderate hypertriglyceridemia (part IC of the clinical study). Percent change from baseline of plasma fasting triglyceride concentrationsand (B) LTW980 plasma concentrations in human subjects with moderate hypertriglyceridemia (part IC) following administration of LTW980 (450 mg s.c.).

[0050] Figure 3 depicts that LTW980 administration lowered postprandial triglycerides in subjects with moderate hypertriglyceridemia (part IC of the clinical study). Percent change from baseline of plasma postprandial triglyceride area under the curve (AUC) following LTW980 administration (450 mg s.c.). AUC was measured during a meal tolerance at baseline and on day 5 after LTW980 administration.

[0051] Figure 4 depicts that triglycerides did not decrease in subjects administered placebo in the elevated triglyceride cohort (IC).

[0052] Figure 5 depicts that in cohort IA, a single subcutaneous dose of LTW980 (450 mg) lowered fasting plasma triglyceride levels (A) with placebo shown in (B).

[0053] Figure 6 depicts that in cohort IB, a single subcutaneous dose of LTW980 (450 mg) lowered mean fasting plasma triglyceride levels (A) with placebo shown in (B).

[0054] Figure 7 depicts that in the elevated triglyceride cohort (part IC), LTW980 administration did not significantly affect plasma LDL cholesterol (A) compared to placebo (B)

[0055] Figure 8 depicts that in the elevated triglyceride cohort (part IC), LTW980 administration resulted in a trend toward increased plasma HDL cholesterol (A) with placebo shown in (B).

[0056] Figure 9 depicts that in the elevated triglyceride cohort (part IC), LTW980 administration did not significantly affect glucose and insulin levels during an oral glucose tolerance test. (A) Glucose. (B) Insulin. Note that timepoints for LTW980 and placebo treated subjects were the same; the offset on the graphs between the groups was added so that the data points and error bars are not overlapping.

[0057] Figure 10 depicts that in the elevated triglyceride cohort (part IC), LTW980 administration did not significantly affect circulating inflammatory biomarkers. (A) hsCRP, (B) SAA, (C) haptoglobin (D) IL-6, (E) IL- 18, (F) CXCL-10, and (G) TNFa. Changes in (H) fasting triglycerides, and (I) percent change from baseline in fasting triglycerides are also shown. Note that for parts (A), (B) and (C) the timepoints for LTW980 and placebo treated subjects were the same; the offset on the graphs between the groups was added so that the data points and error bars are not overlapping.

[0058] Figure 11 depicts pharmacokinetics of a single subcutaneous dose of LTW980 in human subjects. (A) Cohort IA (healthy subjects, doses of 15, 75, 150 and 450 mg). (B) Cohort IB (high BMI subjects). (C) Cohort 1C (high triglyceride participants).

[0059] Figure 12 depicts that LTW980 prevents LPL inhibition by human and cynomolgus monkey ANGPTL4. Data are mean values + / - standard deviation from three replicate wells from a single experiment.

[0060] Figure 13 depicts (A) LTW980 lowered plasma triglycerides in hypertriglyceridemic monkeys. Percent change from day 0 in plasma triglyceride concentrations, (B) LTW980 plasma concentrations in hypertriglyceridemic monkeys administered 3 mg / kg LTW980 by subcutaneous injection, (C) total plasma ApoB, (D) increased HDL-C concentrations, (E) ApoC-III concentrations and increased HDL-C concentrations, (F) FPLC separation TG content per fraction (mg / dL). Percent changes in plasma triglyceride concentrations were calculated for each monkey versus its day 0 value, then group means ± SEM were calculated (n = 4).

[0061] Figure 14 depicts foamy macrophage accumulation in the mesenteric lymph nodes (MLN) of an older, metabolically compromised monkey fed a high-fat diet and given weekly doses of LTW980 for 4 weeks: (A) Micrograph of an MLN. Arrows indicate some of the foamy macrophage aggregates. (B) Magnification of a portion of the image in (A) showing inflammatory cell infiltrate (highlighted with a circle) associated with foamy macrophage accumulation. (C) Electron micrograph of a foamy macrophage showing intracellular lipid droplets.

[0062] Figure 15 depicts that 14D12 treatment lowered fasting plasma triglycerides in DBA / 2 mice.

[0063] Figure 16 depicts mesenteric lymphatic of a DBA / 2 mouse after consuming a high-fat diet and being treated with anti-mouse ANGPTL4 antibody 14D12 for 8 weeks, with microscopic findings of moderate, multifocal aggregates of foamy macrophages with multinucleated giant cells, including Teuton giant cells associated with moderate mixed cell inflammation (yellow star) and fibrosis (yellow triangle) as well as marked vacuolation of foamy macrophages (blue arrows) with multifocal pale eosinophilic acellular material. (B) shows a higher magnification of a portion of the image in (A).

[0064] Figure 17 depicts mesenteric lymph node of a DBA / 2 mouse after consuming a high-fat diet and being treated with anti-mouse ANGPTL4 antibody 14D12 for8 weeks, with minimal multifocal foamy macrophage aggregate in the subcapsular sinus with multinucleate giant cells (blue arrow) and mild vacuolation of foamy macrophages (yellow star) (B) shows a higher magnification of a portion of the image in (A).

[0065] Figure 18 depicts the dose-finding human study design in FPLD-adjacent cohorts. Subjects will be administered a single dose of LTW980 or placebo via subcutaneous injection biweekly for 14 weeks.

[0066] Figure 19 depicts the proof-of-concept human study design in FPLD cohorts. Subjects will be administered a single dose of LTW980 or placebo via subcutaneous injection biweekly for 14 weeks.

[0067] Figure 20 show remnant cholesterol concentrations in Part IC (high triglycerides cohort).

[0068] Figure 21 shows remnant cholesterol in hypertriglyceridemic cynomolgus monkeys after a single dose of LTW890 (3 mg / kg subcutaneous (SC)). Remnant cholesterol (RC) is shown as TRL in the figure. HDL = high-density lipoprotein; LDL = low-density lipoprotein; SC = subcutaneous; TRL = triglyceride-rich lipoprotein.

[0069] Figure 22 shows remnant cholesterol level and insulin sensitivity in mice treated with an exemplary mouse anti-ANGPTL4 antibody (14D12). Treatment with the exemplary mouse anti-ANGPTL4 antibody (14D12) reduced remnant cholesterol levels, fasting insulin, and fed fatty acids.

[0070] Figure 23 is a schematic overview of Lp(a), LDL, IDL, VLDL remnants and chylomi cron-Remnants. HDL = high-density-lipoproteins; IDL = intermediate-density lipoproteins; LDL = low-density-lipoproteins; TG = triglyceride; VLDL = very low-density lipoproteins.

[0071] Figure 24 shows triglyceride / high-density lipoprotein-cholesterol ratio (a surrogate marker for insulin sensitivity) in Part IC (high triglyceride cohort).

[0072] Figure 25 depicts that administration of 300 mg and 450 mg doses of LTW980 substantially lowered (A) triglyceride and (B) remnant cholesterol levels versus placebo treated patients with metabolic dysfunction who had baseline triglyceride levels lower than 150 mg / dL. Percent changes from baseline were calculated for each treated patient versus baseline during active dosing at 6 and 12 weeks and during follow up at 18 and 24 weeks, then group means ± SEM were calculated.

[0073] Figure 26 depicts that administration of 300 mg and 450 mg doses of LTW980 substantially lowered (A) triglyceride and (B) remnant cholesterol levels versus placebo at 12 weeks in the full cohort of patients with metabolic dysfunction who had baseline triglyceride levels lower than 150 mg / dL. Percent changes from baseline were calculated for each treated patient versus baseline, then group means ± SEM were calculated.

[0074] Figure 27 depicts that administration of 300 mg and 450 mg doses of LTW980 substantially lowered (A) triglyceride and (B) remnant cholesterol levels versus placebo at 12 weeks in patients with metabolic dysfunction who had baseline triglyceride levels greater than 150 ml / dL. Percent changes from baseline were calculated for each treated patient versus baseline, then group means ± SEM were calculated.

[0075] Figure 28 depicts that administration of 300 mg and 450 mg doses of LTW980 consistently lowered (A) triglyceride and (B) remnant cholesterol levels versus placebo at 12 weeks in patients with metabolic dysfunction. Percent changes from baseline were calculated versus baseline for all patients with available data at baseline.

[0076] Figure 29 depicts that administration of 150 (n=9), 300 (n=10), and 450 (n=19) mg doses of LTW980 lowers remnant cholesterol levels versus placebo (n=10) in patients who had baseline triglyceride levels greater than 150 ml / dL and 200 ml / dL over 12 weeks of treatment. Percent changes from baseline were calculated for each treatment group versus baseline at 1, 6, and 12 weeks, then group means ± SEM were calculated.

[0077] Figure 30. Percent change from baseline in fasting TG for participants with high TG (Part IC) following a single of LTW980 (450 mg SC),

[0078] Figure 31. Effect of LTW980 on triglyceride and remnant cholesterol (RC) levels through 24 weeks in multidose clinical study. LTW980 treatment resulted in reductions in TG (left) and RC (right) levels that were evident 1 week after treatment began and persisted through 12 weeks of dosing, returning toward baseline after dosing, as observed at the 24-week final visit. Error bars indicate standard error of the mean.

[0079] Figure 32. HDL-C, LDL-C, and TC levels with single-dose LTW980 treatment in Part IC cohort in first-in-human clinical study.

[0080] Figure 33. Serum inflammatory biomarker levels with single-dose LTW980 treatment in Part IC cohort of first-in-human clinical study.

[0081] Figure 34. VLDL, HDL-C, non-HDL-C, and LDL-C levels with LTW980 treatment in multidose clinical study.

[0082] Figure 35. Serum inflammatory biomarker levels with LTW980 treatment in multi-dose clinical study.

[0083] Figure 36. HbAlc, HOMA-IR fasting glucose and fasting insulin levels with LTW980 treatment in multidose clinical study.

[0084] Figure 37. LTW980 lowers lipoprotein TG and cholesterol content derived from FPLC. Traces showing FPLC-derived lipoprotein TG and cholesterol content from remaining 3 animals treated with 3 mg / kg LTW980. Cholesterol content in TRL (b) is RC. (FPLC = fast performance liquid chromatography; min = minute; TG = triglycerides; TRL = triglyceride-rich lipoprotein; RC = remnant cholesterol).

[0085] Figure 38. Serum inflammatory biomarker levels in NHPs on HSFD administered LTW980 for 15 weeks a) hsCRP in males b) hsCRP in females c) SAA in males d) SAA in females e) fibrinogen in males f) fibrinogen in females. Figures represent plotted values of Supplementary Table 2. Values below the lower limit of quantification (BLoQ) were assigned as 0 5 x LLoQ (lower limit of quantification) and those ALoQ (above lower limit of quantification) were assigned as 1 -5 x ULoQ (upper limit of quantification) for calculations. Accordingly, some biomarkers may have 0 SD if all samples were BLoQ or ALoQ. indicates biomarker was not measured at the timepoint. (M = Male; F = Female; SD = standard deviation; hsCRP = high-sensitivity C-reactive Protein; SAA = Serum amyloid A; FIB = Fibrinogen; HSFD = high saturated fat diet; Pl = pre-dose timepoint 1; P2 = pre-dose timepoint 2; D28,56,98 = Days 28, 56, and 98, respectively).

[0086] Figure 39. Serum concentrations of LTW980 7 days after administration of the first dose (Week 1) and at the end of the dose interval (Week 12). Bars represent mean serum LTW980 concentrations for each LTW980 dosing group (150, 300, or 450 mg). Error bars represent standard deviation.

[0087] Figure 40. Evaluation of LTW980 target-mediated clearance in NHPs. A) serum concentration-time profiles of LTW980 at doses of 1 mg / kg, 3 mg / kg, or 30 mg / kg in cynomolgus monkeys. B) LTW980 clearance rate in cynomolgus monkeys. C) Curve fitting of Figure 40 A based on a compartmental model that includes linear and nonlinear clearance. D) Predicted serum concentration-time profiles for LTW980 at doses of 1, 3, and 10 mg / kg over 30 days. E) Simulated serum concentration-time curves of LTW980 at doses of 1, 3, 6,10, 15, or 20 mg / kg. F) Schematic depicting that target-mediated clearance diminishes at higher doses as linear clearance dominates. G) Two-compartment model to depict total clearance as sum of target and linear clearance components.

[0088] Figure 41. Comparison of LTW980 clearance in humans and NHPs. A) Estimates value comparison of dose-normalized AUC for LTW980 and clearance (CL / F) in healthy subjects. B) Human clearance rate. C) Comparison of human and NHP clearance rates.

[0089] Figure 42. Comparison of pharmacokinetics of LTW980 between NHP (A) and human (B) based on dose-normalized AUC (AUC / Dose) and (C) species-specific saturation thresholds following subcutaneous dosing.

[0090] Figure 43. Proposed pharmacokinetic study design for LTW980-HLE (halflife extended (HLE) version of LTW980 described herein) in naive cynomolgus monkeys.

[0091] Figure 44. Simulated serum concentration-time profiles of LTW980-HLE compared to the parent construct (LTW980) at 1 mg / kg and 10 mg / kg doses over 60 days.

[0092] Figure 45. Schematic of Phase 2b Randomized, Double-Blind, Parallel - Group, Placebo-Controlled study design.DEFINITIONS

[0093] In order for the present disclosure to be more readily understood, certain terms are first defined below. Additional definitions for the following terms and other terms are set forth throughout the specification. The publications and other reference materials referenced herein to describe the background of the disclosure and to provide additional detail regarding its practice are hereby incorporated by reference.

[0094] The articles “a” and “an” are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an agent” means one agent or more than one agent.

[0095] About'. As used herein, the term “about,” as applied to one or more values of interest, refers to a value that is similar to a stated reference value. In some embodiments, the term “about” refers to a range of values that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1 %, or less ineither direction (greater than or less than) of the stated reference value unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value).

[0096] Agent'. As used herein, the term “agent” refers to a biological entity and / or compound including, for example, an antibody or antigen-binding fragment thereof, an organic molecule (e.g., a small molecule), a peptide (e.g., a fusion protein), an aptamer, a nucleic acid, a chimeric antigen receptor, a glycoprotein, a saccharide, a lipid, a growth factor, an enzyme, a synthetic molecule, a carbohydrate, a lipid, a hormone, a polymer, or a derivative, variation, complex, or any combination thereof. In appropriate circumstances, as will be clear from context to those skilled in the art, the term may be utilized to refer to an entity that is or comprises a cell or organism, or a fraction, extract, or component thereof. Alternatively or additionally, as context will make clear, the term may be used to refer to a natural product. In some instances, again as will be clear from context, the term may be used to refer to one or more entities that is man-made in that it is designed, engineered, and / or produced through human action and / or is not found in nature. In some embodiments, an agent may be utilized in isolated or pure form. In some embodiments, an agent may be utilized in crude form. In some embodiments, agents are provided as collections or libraries, which may be screened to identify or characterize active agents within them. An agent may bind any cell moiety, such as a receptor, an antigenic determinant, or other binding site present on a target or target cell. Various agents are useful in the compositions and methods described herein.

[0097] ANGPTL4, ANGPTL4 protein or ANGPTL4 antigen'. As used herein, the terms “ANGPTL4”, “ANGPTL4 protein” or “ANGPTL4 antigen” are used interchangeably, and refer to the Angiopoietin-like 4 (ANGPTL4) protein in different species. For example, human ANGPTL4 has the sequence as set out in Table 1 (SEQ ID NO: 1). It is a homooligomeric protein, capable of forming dimers and tetramers. In addition, in the context of this disclosure, the term “ANGPTL4” includes mutants of the natural Angiopoietin-like 4 (ANGPTL4) protein, which have substantially the same amino acid sequence as that of the native primary structure (amino acid sequence) described in the above-mentioned reports. Herein, the term “mutants of the natural human Angiopoietin-like 4 (ANGPTL4) protein having substantially the same amino acid sequence” refers to such mutant proteins.

[0098] ANGPTL4-associated disease, disorder, or condition'. As used herein, the term “ANGPTL4-associated disease, disorder, or condition” refers to any number of diseases, disorders or conditions in which a reduction of ANGPTL4-mediated LPL inhibition and lipoprotein modulation is sought. These conditions include but are not limited to those involving lipid metabolism, such as hypercholesterolemia, hypertriglyceridemia, hyperlipidemia, hyperlipoproteinemia and dyslipidemia or lipodystrophy. In some embodiments, lipodystrophy includes, but is not limited to, familial lipodystrophy or acquired lipodystrophy. In some embodiments, dyslipidemia includes, but is not limited to, atherogenic dyslipidemia, diabetic dyslipidemia, mixed dyslipidemia, hypertriglyceridemia, hypercholesterolemia, chylomicronemia, and / or other conditions caused by, e.g., decreased LPL activity and / or LPL deficiency, decreased LDL receptor activity and / or LDL receptor deficiency, altered ApoC2, ApoE deficiency, increased ApoB, increased production and / or decreased elimination of very low-density lipoprotein (VLDL), certain drug treatment (e.g., glucocorticoid treatment-induced dyslipidemia), any genetic predisposition, diet, lifestyle, and the like. In some embodiments, hypertriglyceridemia includes, but is not limited to, severe hypertriglyceridemia (e.g., with plasma triglyceride concentration >500 mg / dL), hypertriglyceridemia associated with obesity, and type V hypertriglyceridemia.

[0099] In some embodiments, ANGPTL4-associated disease, disorder, or condition may be associated with or resulting from one or more additional disease, disorder, or condition. Other ANGPTL4-associated diseases, disorders, or conditions associated with or resulting from e.g., hyperlipidemia, hyperlipoproteinemia, and / or dyslipidemia, include, but are not limited to, obesity, fatty liver disease (e.g., nonalcoholic fatty liver disease), insulin resistance, metabolic syndrome, metabolic dysfunction, polycystic ovarian syndrome, chronic kidney disease, Cushing syndrome, hypercorti soli sm, acromegaly, heart failure, cardiovascular diseases or disorders, acute pancreatitis, nonalcoholic steatohepatitis (NASH); blood sugar disorders;, and the like. In some embodiments, cardiovascular diseases or disorders include, but are not limited to, atherosclerosis, aneurysm, hypertension, angina, stroke, cerebrovascular diseases, congestive heart failure (e.g., heart failure with preserved ejection fraction), coronary artery diseases, myocardial infarction, peripheral vascular diseases, and the like. In some embodiments, bloods sugar disorders include, but are not limited to, diabetes (e.g., type 2 diabetes).

[0100] Antibody. As used herein, the term “antibody” refers to a polypeptide that includes canonical immunoglobulin sequence elements sufficient to confer specific binding to a particular target antigen. As is known in the art, intact antibodies as produced in nature are approximately 150 kD tetrameric agents comprising two identical heavy chain polypeptides (about 50 kD each) and two identical light chain polypeptides (about 25 kD each) that associate with each other into what is commonly referred to as a “Y-shaped” structure. Each heavy chain comprises at least four domains (each about 110 amino acids long) - an amino-terminal variable (VH) domain (located at the tips of the Y structure), followed by three constant domains: CHI, CH2, and the carboxy-terminal CH3 (located at the base of the Y’s stem). A short region, known as the “switch,” connects the heavy chain variable and constant regions. The “hinge” connects CH2 and CH3 domains to the rest of the antibody. Two disulfide bonds in this hinge region connect the two heavy chain polypeptides to one another in an intact antibody. Each light chain comprises two domains - an amino-terminal variable (VL) domain, followed by a carboxy-terminal constant (CL) domain, separated from one another by another “switch”. Intact antibody tetramers comprise two heavy chain-light chain dimers in which the heavy and light chains are linked to one another by a single disulfide bond; two other disulfide bonds connect the heavy chain hinge regions to one another so that the dimers are connected to one another and a tetramer is formed. Naturally-produced antibodies are also glycosylated, typically on the CH2 domain. Each domain in a natural antibody has a structure characterized by an “immunoglobulin fold” formed from two beta sheets (e.g., 3-, 4-, or 5-stranded sheets) packed against each other in a compressed antiparallel beta barrel.

[0101] Each variable domain contains three hypervariable loops known as “complementarity determining regions” (CDR1, CDR2, and CDR3) and four somewhat invariant “framework” regions (FR1, FR2, FR3, and FR4). In some embodiments, an ANGPTL4 binding antibody or antigen-binding fragment thereof is or comprises a polypeptide whose amino acid sequence includes at least one CDR (e.g., at least one heavy chain CDR and / or at least one light chain CDR) that is substantially identical to one found in a reference antibody. In some embodiments, an included CDR substantially identical to a reference CDR in that it is either identical in sequence or contains between 1-5 amino acid substitutions as compared with the reference CDR. In some embodiments, an included CDR is substantially identical to a reference CDR in that at least one amino acid within theincluded CDR is deleted, added, or substituted as compared with the reference CDR, but the included CDR has an amino acid sequence that is otherwise identical with that of the reference CDR. In some embodiments an included CDR is substantially identical to a reference CDR in that 1-5 amino acids within the included CDR are deleted, added, or substituted as compared with the reference CDR, but the included CDR has an amino acid sequence that is otherwise identical to the reference CDR. In some embodiments, an included CDR is substantially identical to a reference CDR in that at least one amino acid within the included CDR is substituted as compared with the reference CDR, but the included CDR has an amino acid sequence that is otherwise identical with that of the reference CDR. In some embodiments, an included CDR is substantially identical to a reference CDR in that 1-5 amino acids within the included CDR are deleted, added, or substituted as compared with the reference CDR, but the included CDR has an amino acid sequence that is otherwise identical to the reference CDR.

[0102] In some embodiments, an ANGPTL4 binding antibody or antigen-binding fragment thereof is or comprises a polypeptide whose amino acid sequence includes structural elements recognized by those skilled in the art as an immunoglobulin variable domain. In some embodiments, an ANGPTL4 binding antibody or antigen-binding fragment thereof is a polypeptide protein having a binding domain that is homologous or largely homologous to an immunoglobulin-binding domain. When natural antibodies fold, the FR regions form the beta sheets that provide the structural framework for the domains, and the CDR loop regions from both the heavy and light chains are brought together in three-dimensional space so that they create a single hypervariable antigen binding site located at the tip of the Y structure. The Fc region of naturally-occurring antibodies binds to elements of the complement system, and also to receptors on effector cells, including, for example, effector cells that mediate cytotoxicity. Affinity and / or other binding attributes of Fc regions for Fc receptors can be modulated through glycosylation or other modification. In some embodiments, antibodies produced and / or utilized in accordance with the present disclosure include glycosylated Fc domains, such as Fc domains with modified or engineered glycosylation. In some embodiments, any polypeptide or complex of polypeptides that includes sufficient immunoglobulin domain sequences as found in natural antibodies can be referred to and / or used as an “antibody”, whether such polypeptide is naturally produced (e.g., generated by an organism reacting to an antigen) or produced byrecombinant engineering, chemical synthesis, or other artificial system or methodology. In some embodiments, an antibody is polyclonal. In some embodiments, an antibody is monoclonal. In some embodiments, an antibody has constant region sequences characteristic of mouse, rabbit, primate, or human antibodies. In some embodiments, antibody sequence elements are humanized, primatized, or chimeric as is known in the art.

[0103] Moreover, the term “antibody,” as used herein, can refer in appropriate embodiments (unless otherwise stated or clear from context) to any of the art-known or developed constructs or formats for utilizing antibody structural and functional features in alternative presentation. For example, in some embodiments, an antibody utilized in accordance with the present disclosure is in a format selected from, but not limited to, intact IgA, IgG, IgE or IgM antibodies; bispecific or multi- specific antibodies (e.g., Zybodies®, etc); and / or antibody fragments (preferably antibody fragments that exhibit desired antigenbinding activity). An antibody described herein can be an immunoglobulin, heavy chain antibody, light chain antibody, LRR-based antibody, or other protein scaffold with antibodylike properties, as well as any other immunological binding moiety known in the art, e.g., a Fab, Fab', Fab'2, Fab2, Fab3, F(ab’)2 , Fd, Fv, Feb, scFv, SMIP, antibody, diabody, triabody, tetrabody, minibody, maxibody, tandab, DVD, BiTe, TandAb, or any combination thereof. The subunit structures and three-dimensional configurations of different classes of antibodies are known in the art. In some embodiments, an antibody may lack a covalent modification (e.g., attachment of a glycan) that it would have if produced naturally. In some embodiments, an antibody may contain a covalent modification (e.g., attachment of a glycan, a payload (e.g., a detectable moiety, a therapeutic moiety, a catalytic moiety, etc), or other pendant group (e.g., poly-ethylene glycol, etc.).

[0104] Antibody heavy chain'. As used herein, the term “antibody heavy chain” refers to the larger of the two types of polypeptide chains present in all antibodies in their naturally occurring conformations.

[0105] Antibody light chain'. As used herein, the term “antibody light chain” refers to the smaller of the two types of polypeptide chains present in all antibodies in their naturally occurring conformations.

[0106] Antigen'. As used herein, the term “antigen” or “Ag” refers to a molecule that is capable of provoking an immune response. This immune response may involve either antibody production, the activation of specific immunologically-competent cells, or both. Askilled artisan will understand that any macromolecule, including virtually all proteins or peptides, can serve as an antigen. Furthermore, antigens can be derived from recombinant or genomic DNA. A skilled artisan will understand that any DNA that comprises a nucleotide sequence or a partial nucleotide sequence encoding a protein that elicits an immune response encodes an “antigen” as that term is used herein. Furthermore, one skilled in the art will understand that an antigen need not be encoded solely by a full length nucleotide sequence of a gene. It is readily apparent that the present disclosure includes, but is not limited to, the use of partial nucleotide sequences of more than one gene and that these nucleotide sequences are arranged in various combinations to elicit the desired immune response. Moreover, a skilled artisan will understand that an antigen need not be encoded by a “gene” at all. It is readily apparent that an antigen can be generated synthesized or can be derived from a biological sample. Such a biological sample can include, but is not limited to a tissue sample, a tumor sample, a cell, or a biological fluid.

[0107] Antigen-binding fragment. As used herein, the term “antigen-binding fragment” refers to a portion of an intact antibody that binds the antigen to which the intact antibody binds. An antigen-binding fragment of an antibody includes any naturally occurring, enzymatically obtainable, synthetic, or genetically engineered polypeptide or glycoprotein that specifically binds an antigen to form a complex. Exemplary antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab')2; diabodies; linear antibodies; single-chain antibody molecules (e.g., scFv, VHH, cam elid, or VH or VL domains only); or multispecific antibodies formed from antibody fragments. In some embodiments, the antigen-binding fragments of the antibodies described herein are scFvs. In some embodiments, the antigen-binding fragments of the antibodies described herein are VHH domains only. As with full antibody molecules, antigen-binding fragments may be mono-specific or multispecific (e.g., bispecific). A multispecific antigen-binding fragment of an antibody may comprise at least two different variable domains, wherein each variable domain is capable of specifically binding to a separate antigen or to a different epitope of the same antigen. An antigen-binding fragment may be produced by any means. For example, in some embodiments, an antigen-binding fragment is enzymatically or chemically produced by fragmentation of an intact antibody. Alternatively, in some embodiments, an antigenbinding fragment is recombinantly produced. In some embodiments, an antigen-binding fragment is wholly or partially synthetically produced. In some embodiments, an antigen-binding fragment has a length of at least about 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 amino acids or more.

[0108] Associated. Two events or entities are “associated” with one another, as that term is used herein, if the presence, level, degree, type and / or form of one is correlated with that of the other. For example, a particular entity (e.g., polypeptide, genetic signature, metabolite, or microbe) is considered to be associated with a particular disorder, if its presence, level and / or form correlates with incidence of and / or susceptibility to the disorder (e.g., across a relevant population). In some embodiments, two or more entities are physically “associated” with one another if they interact, directly or indirectly, so that they are and / or remain in physical proximity with one another. In some embodiments, two or more entities physically associated with one another are covalently linked to one another. In some embodiments, two or more entities physically associated with one another are not covalently linked to one another but are non-covalently associated, for example, by means of hydrogen bonds, van der Waals interaction, hydrophobic interactions, magnetism, and combinations thereof.

[0109] Binding'. As used herein, the term “binding” refers to a non-covalent association between or among two or more entities. “Direct” binding involves physical contact between entities or moieties. Indirect binding involves physical interaction by way of physical contact with one or more intermediate entities. Binding between two or more entities can typically be assessed in any of a variety of contexts - including where interacting entities or moieties are studied in isolation or in the context of more complex systems (e.g., while covalently or otherwise associated with a carrier entity and / or in a biological system or cell).

[0110] Carrier, as used herein, “carrier” refers to a diluent, adjuvant, excipient, and / or vehicle with which a composition is administered. In some exemplary embodiments, carriers include sterile liquids, such as, for example, water and oils, including oils of petroleum, animal, vegetable or synthetic origin, such as, for example, peanut oil, soybean oil, mineral oil, sesame oil and the like. In some embodiments, carriers are or include one or more solid components.

[0111] CDR. As used herein, “CDR” refers to a complementarity determining region within an antibody variable region. There are three CDRs in each of the variable regions of the heavy chain and the light chain, which are designated CDR1, CDR2 andCDR3, for each of the variable regions. A “set of CDRs” or “CDR set” refers to a group of three or six CDRs that occur in either a single variable region capable of binding the antigen or the CDRs of cognate heavy and light chain variable regions capable of binding the antigen. In general, there are three CDRs in each heavy chain variable region (HCDR1, HCDR2, HCDR3) and three CDRs in each light chain variable region (LCDR1, LCDR2, LCDR3). The exact definitional CDR boundaries and lengths are subject to different classification and numbering systems. Certain systems have been established in the art for defining CDR boundaries (e.g., Kabat, IMGT, Chothia, or a combination thereof). CDRs may therefore be referred to by Kabat, Chothia, IMGT, or any other boundary definitions known in the art. Despite differing boundaries, each of these systems has some degree of overlap in what constitutes the “hypervariable regions” within the variable sequences. CDR definitions according to these systems may therefore differ in length and boundary areas with respect to the adjacent framework region (see, e.g., Kabat et al., in “Sequences of Proteins of Immunological Interest,” 5th Edition, U.S. Department of Health and Human Services, 1992; Chothia et al. (1987) J. Mol. Biol. 196, 901; and MacCallum et al., J. Mol. Biol. (1996) 262, 732, each of which is incorporated by reference in its entirety). Those skilled in the art appreciate the differences between and among these systems and are capable of understanding CDR boundaries to the extent required to understand and to practice the claims and disclosure herein.

[0112] Composition'. Those skilled in the art will appreciate that the term “composition” may be used to refer to a discrete physical entity that comprises one or more specified components. In general, unless otherwise specified, a composition may be of any form - e.g., gas, gel, liquid, or solid.

[0113] Comprising'. A composition or method described herein as “comprising” one or more named elements or steps is open-ended, meaning that the named elements or steps are essential, but other elements or steps may be added within the scope of the composition or method. To avoid prolixity, it is also understood that any composition or method described as "comprising” (or which “comprises”) one or more named elements or steps also describes the corresponding, more limited composition or method “consisting essentially of’ (or which "consists essentially of') the same named elements or steps, meaning that the composition or method includes the named essential elements or steps and may also include additional elements or steps that do not materially affect the basic and novel characteristic(s)of the composition or method. It is also understood that any composition or method described herein as “comprising” or “consisting essentially of’ one or more named elements or steps also describes the corresponding, more limited, and closed-ended composition or method “consisting of’ (or “consists of’) the named elements or steps to the exclusion of any other unnamed element or step. In any composition or method disclosed herein, known or disclosed equivalents of any named essential element or step may be substituted for that element or step.

[0114] Combination therapy. The term “combination therapy”, as used herein, refers to those situations in which a subject is simultaneously exposed to two or more therapeutic regimens (e.g., two or more therapeutic agents). In some embodiments, a therapeutic regimen is a specific diet (e.g., low carbohydrate diet) exercise schedule etc. In some embodiments, it refers to those situations in which two or more different therapeutic agents are administered in overlapping regimens so that the subject is simultaneously exposed to both agents. When used in combination therapy, two or more different therapeutic agents may be administered simultaneously or separately. This administration in combination can include simultaneous administration of the two or more therapeutic agents in the same dosage form, simultaneous administration in separate dosage forms, and separate administration. That is, two or more therapeutic agents can be formulated together in the same dosage form and administered simultaneously. Alternatively, two or more therapeutic agents can be simultaneously administered, wherein the agents are present in separate formulations. In another alternative, a first therapeutic agent can be administered followed by one or more additional therapeutic agents. In the separate administration protocol, two or more therapeutic agents may be administered a few minutes apart, or a few hours apart, a few days apart, or a few weeks apart. In some embodiments, two or more therapeutic agents may be administered within hours (e.g., less than about 1 hour, about 2 hours, about 3 hours, about 4 hours, or about 5 hours) apart.

[0115] Dosage unit or dose'. As used herein, a “dosage unit” or “dose” refers to physically discrete units suited as unitary dosages for the particular subject to be treated, e.g., human subject. Each unit can contain a predetermined quantity of active compound(s) calculated to produce the desired therapeutic effect(s) in association with a pharmaceutical carrier. The specification for the dosage unit forms can be dictated by (a) the unique characteristics of the active compound(s) and the particular therapeutic effect(s) to beachieved, and (b) the limitations inherent in the art of compounding such active compound(s).

[0116] Fragment'. As used herein, the term “fragment” refers to a structure that includes a discrete portion of the whole, but lacks one or more moieties found in the whole structure. In some embodiments, a fragment consists of such a discrete portion. In some embodiments, a fragment consists of or comprises a characteristic structural element or moiety found in the whole. In some embodiments, an antigen-binding fragment comprises or consists of at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, or more monomeric units (e.g., amino acids) as found in a whole antibody. In some embodiments, an antigen-binding fragment comprises or consists of at least about 5%, 10%, 15%, 20%, 25%, 30%, 25%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more of the monomeric units (e.g., residues) found in a whole antibody. In some embodiments, a nucleotide fragment comprises or consists of at least about 5%, 10%, 15%, 20%, 25%, 30%, 25%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more of the monomeric units (e.g., residues) found in the whole nucleotide.

[0117] Identity. As used herein, the term “identity” refers to the subunit sequence identity between two polymeric molecules, particularly between two amino acid molecules, such as between two polypeptide molecules. When two amino acid sequences have the same residues at the same positions; e.g., if a position in each of two polypeptide molecules is occupied by an Arginine, then they are identical at that position. The identity or extent to which two amino acid sequences have the same residues at the same positions in an alignment is often expressed as a percentage. The identity between two amino acid sequences is a direct function of the number of matching or identical positions; e.g., if half of the positions (e.g., five positions in a polymer of 10 amino acids in length) in two sequences are identical, the two sequences are 50% identical; if 90% of the positions (e.g., nine positions in a polymer of 10 amino acids in length) are identical, the two amino acids sequences are 90% identical.

[0118] “Improve, ” “increase ”, “inhibit” o “reduce ”'. As used herein, the terms “improve,” “increase,” “inhibit,” “reduce,” or grammatical equivalents thereof, indicatevalues that are relative to a baseline or other reference measurement. In some embodiments, an appropriate reference measurement is or comprises a measurement in a particular system (e.g., in a single subject) under otherwise comparable conditions absent presence of (e.g., prior to and / or after) a particular agent or treatment, or in presence of an appropriate comparable reference agent. In some embodiments, an appropriate reference measurement is or comprises a measurement in comparable system known or expected to respond in a particular way, in presence of the relevant agent or treatment.

[0119] Nucleic acid. As used herein, the term “nucleic acid” refers to a polymer of at least three nucleotides. In some embodiments, a nucleic acid comprises DNA. In some embodiments, a nucleic acid comprises RNA. In some embodiments, a nucleic acid is single stranded. In some embodiments, a nucleic acid is double stranded. In some embodiments, a nucleic acid comprises both single and double stranded portions. In some embodiments, a nucleic acid comprises a backbone that comprises one or more phosphodiester linkages. In some embodiments, a nucleic acid comprises a backbone that comprises both phosphodiester and non-phosphodiester linkages. For example, a nucleic acid may comprise a backbone that comprises one or more phosphorothioate or 5'-N-phosphoramidite linkages and / or one or more peptide bonds, e.g., as in a peptide nucleic acid. In some embodiments, a nucleic acid comprises one or more, or all, natural residues (e.g., adenine, cytosine, deoxyadenosine, deoxycytidine, deoxyguanosine, deoxythymidine, guanine, thymine, and / or uracil). In some embodiments, a nucleic acid comprises one or more, or all, non-natural residues. In some embodiments, a non-natural residue comprises a nucleoside analog (e.g., 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3 -methyl adenosine, 5- methylcytidine, C-5 propynyl-cytidine, C-5 propynyl-uridine, 2-aminoadenosine, C5- bromouridine, C5-fluorouridine, C5-iodouridine, C5-propynyl-uridine, C5-propynyl- cytidine, C5-methylcytidine, 2-aminoadenosine, 7-deazaad enosine, 7-deazaguanosine, 8- oxoadenosine, 8-oxoguanosine, O(6)-methylguanine, 2-thiocytidine, methylated bases, intercalated bases, or combinations thereof). In some embodiments, a non-natural residue comprises one or more modified sugars (e.g., 2'-fluororibose, ribose, 2'-deoxyribose, arabinose, and / or hexose) as compared to those in natural residues. In some embodiments, a nucleic acid has a nucleotide sequence that encodes a functional gene product, such as an RNA or polypeptide. In some embodiments, a nucleic acid has a nucleotide sequence that comprises one or more introns. In some embodiments, a nucleic acid may be prepared byisolation from a natural source, enzymatic synthesis (e.g., by polymerization based on a complementary template, e.g., in vivo or in vitro, reproduction in a recombinant cell or system, or chemical synthesis. In some embodiments, a nucleic acid is at least 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 20, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 600, 700, 800, 900, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000 or more residues long.

[0120] Pharmaceutically acceptable '. As used herein, the term “pharmaceutically acceptable” refers to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable carrier '. As used herein, the term “pharmaceutically acceptable carrier” means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, or solvent encapsulating material, involved in carrying or transporting the subject compound from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulation and not injurious to the subject. Some examples of materials which can serve as pharmaceutically acceptable carriers include sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, com oil and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer’s solution; ethyl alcohol; pH buffered solutions; polyesters, polycarbonates and / or polyanhydrides; and other non-toxic compatible substances employed in pharmaceutical formulations.Substantially. As used herein, the term “substantially” refers to the qualitative condition of exhibiting total or near-total extent or degree of a characteristic or property of interest. One of ordinary skill in the biological arts will understand that biological and chemicalphenomena rarely, if ever, go to completion and / or proceed to completeness or achieve or avoid an absolute result. The term “substantially” is therefore used herein to capture the potential lack of completeness inherent in many biological and chemical phenomena.

[0121] Treat'. As used herein, the terms “treat,” “treatment,” or “treating” refer to partial or complete alleviation, amelioration, delay of onset of, inhibition, prevention, relief, and / or reduction in incidence and / or severity of one or more symptoms or features of a disease, disorder, and / or condition. In some embodiments, treatment is administered to a subject who does not exhibit signs or features of a disease, disorder, and / or condition (e.g., may be prophylactic). In some embodiments, treatment is administered to a subject who exhibits only early or mild signs or features of the disease, disorder, and / or condition, for example for the purpose of decreasing the risk of developing pathology associated with the disease, disorder, and / or condition. In some embodiments, treatment is administered to a subject who exhibits established, severe, and / or late-stage signs of the disorder. In some embodiments, treating comprises administering at least one ANGPTL4 binding antibody or antigen-binding fragment thereof described herein to a subject.

[0122] Throughout this disclosure, various aspects of the invention can be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the breadth of the range.DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS

[0123] Disclosed herein are methods of treating a subject (e.g., a human) having a disorder with an antibody or antigen-binding fragment thereof that binds specifically to human ANGPTL4. In some embodiments, a disorder is an ANGPTL4-associated disorder.Disclosed are also compositions comprising an antibody or antigen-binding fragment thereof that binds specifically to human ANGPTL4 (e.g., an anti-ANGPTL4 antibody).

[0124] Before the present methods and compositions are described, it is to be understood that this disclosure is not limited to particular method or composition described, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present disclosure will be limited only by the appended claims.ANGPTL4

[0125] The present disclosure provides, among other things, antibodies or antigenbinding fragments thereof that specifically bind to ANGPTL4 (e.g., ANGPTL4 binding antibodies and antigen-binding fragments) and inhibit its biological activities, including but not limited to ability to activate lipoprotein lipase (LPL).

[0126] Angiopoietin-like 4 protein (ANGPTL4) is a member of the angiopoietin family of secreted proteins. ANGPTL4 is expressed mainly in adipose tissue, liver and macrophages, and increases triglyceride and remnant cholesterol (RC) levels by binding to and inhibiting LPL (LPL-mediated processing of triglyceride-rich lipoproteins (TRL) is a rate-limiting step in the clearance of triglycerides and TRL from circulation). ANGPTL4 is also known as hepatic fibrinogen / angiopoietin-related protein (HF ARP) (Kim et al. (2000) Biochem. J. 346:603-610); PPAR gamma angiopoietin related protein (PGAR) (Yoon, et al. (2000) Mol. Cell Biol., 20:5343-5349), and fasting induced adipose factor (FIAF) (Kerten et al. (2000) J. Biol. Chem., 275:28488-28493). ANGPTL4 contains an N-terminal coiled-coil domain and a C-terminal fibrinogen (FBN)-like domain (Kim et al. (2000) Biochem. J. 346:603-610).

[0127] Human carriers of an ANGPTL4 loss-of-function variant (E40K) have lower plasma triglyceride levels and lower atherosclerotic cardiovascular disease (ASCVD) risk, suggesting that ANGPTL4 inhibition could lower triglycerides and ASCVD risk in people with hypertriglyceridemia. Furthermore, E40K loss-of-function alleles in the human population greatly decrease or abolish the ability of ANGPTL4 to inhibit LPL and have been associated with beneficial metabolic effects and reduction of cardiometabolic outcomes. People who are homozygous carriers of ANGPTL4 E40K appear healthy, which suggeststhat the mesenteric inflammatory response seen in mice following ANGPTL4 loss or inhibition (together with a high-fat diet) would not occur in humans following treatment with an ANGPTL4 inhibitor. To assess the therapeutic potential of ANGPTL4 inhibition, we developed a humanized ANGPTL4 antibody (LTW980) and set out to determine its ability to lower plasma triglycerides in human subjects with hypertriglyceridemia.

[0128] Lipoprotein lipase (LPL) has a central role in lipoprotein metabolism to maintain normal lipoprotein levels in blood and, through tissue specific regulation of its activity, to determine when and in what tissues triglycerides (TG) and / or remnant cholesterol are unloaded. The coiled-coil region of ANGPTL4 is known to inhibit lipoprotein lipase (LPL) -mediated triglyceride (TG) and remnant cholesterol clearance. Therefore, ANGPTL4 loss-of-function mutations (e.g., as seen in human subjects), genetic deletions (e.g., as seen in transgenic mice), and antibody inhibition (e.g., as seen in mice and cynomolgus monkeys) are all observed to decrease plasma triglycerides and reduction in cardiovascular risk. Furthermore, ANGPTL4 antibodies are also known to activate LPL.

[0129] The ANGPTL4 antibodies and antigen-binding fragments thereof provided in this disclosure initiate, promote, or enhance activation of LPL, e.g., by blocking ANGPTL4 inhibition of LPL, thereby decreasing plasma triglycerides. In some embodiments, ANGPTL4 antibodies or antigen-binding fragments thereof inhibits ANGPTL4 activity leading to increased metabolism of triglyceride-rich lipoparticles (TRL) and reduction in remnant cholesterol (RC) levels. In some embodiments, antibodies or antigen-binding fragments thereof described herein prevent and ameliorate the acute and chronic manifestations of diseases characterized by elevated triglyceride levels, e.g., lipodystrophy, primary dyslipidemia, hypertriglyceridemia, metabolic dysfunction, metabolic syndrome, type II diabetes, and the like. In some embodiments, antibodies or antigen-binding fragments thereof described herein prevent and ameliorate cardiovascular diseases, such as atherosclerotic cardiovascular disease.ANGPTL4 antibody or antigen-binding fragment thereof

[0130] The present disclosure provides, among other things, antibodies or antigenbinding fragments thereof that specifically bind to ANGPTL4. In some embodiments, the present disclosure provides antibodies or antigen-binding fragments thereof that specifically bind to human ANGPTL4. Antibodies or antigen-binding fragments thereof of thedisclosure include, but are not limited to, humanized ANGPTL4 binding antibodies or antigen-binding fragments thereof as described in the Examples.

[0131] The present disclosure provides antibodies or antigen-bindings fragments thereof that specifically bind to ANGPTL4, wherein the antibodies or antigen-binding fragments thereof comprise a VH CDR having an amino acid sequence of any one of the VH CDRs listed in Table 1, infra. In particular, the disclosure provides antibodies or antigenbinding fragments thereof that specifically bind to ANGPTL4, wherein the antibodies or antigen-binding fragments thereof comprise (or alternatively, consist of) one, two, three, or more VH CDRs having an amino acid sequence of any of the VH CDRs listed in Table 1, infra. The present disclosure also provides antibodies or antigen-bindings fragment thereof that specifically bind ANGPTL4, wherein the antibodies or antigen-binding fragments thereof comprise a VH domain having an amino acid sequence of SEQ ID NOs: 13, 38, 58, 78, 98, 118, and 138.

[0132] The present disclosure provides antibodies or antigen-binding fragments thereof that specifically bind to an ANGPTL4, said antibodies or antigen-binding fragments thereof comprising a VL CDR having an amino acid sequence of any one of the VL CDRs listed in Table 1, infra. In particular, the disclosure provides antibodies or antigen-binding fragments thereof that specifically bind to ANGPTL4, said antibodies or antigen-binding fragments thereof comprising (or alternatively, consisting of) one, two, three or more VL CDRs having an amino acid sequence of any of the VL CDRs listed in Table 1, infra. The present disclosure also provides antibodies or antigen-binding fragments thereof that specifically bind to ANGPTL4, said antibodies or antigen-binding fragments thereof comprising a VL domain having an amino acid sequence of SEQ ID NOs: 23, 48, 68, 88, 108, 128, and 148.

[0133] The present disclosure provides antibodies or antigen-binding fragments thereof that specifically bind to ANGPTL4, wherein the antibodies or antigen-binding fragments thereof comprise a VH CDR having an amino acid sequence of any one of the VH CDRs of SEQ ID NOs: 7, 8, 9, 10, 11, and / or 12. In particular, the disclosure provides antibodies or antigen-binding fragments thereof that specifically bind to ANGPTL4 (e.g., human ANGPTL4), wherein the antibodies or antigen-binding fragments thereof comprise (or alternatively, consist of) one, two, three, or more VH CDRs having an amino acid sequence of any of the VH CDRs of SEQ ID NOs: 7, 8, 9, 10, 11, and / or 12. The presentdisclosure also provides antibodies or antigen-binding fragments thereof that specifically bind ANGPTL4 (e.g., human ANGPTL4), wherein the antibodies or antigen-binding fragments thereof comprise a VH domain having an amino acid sequence of SEQ ID NOs: 13 and / or 25.

[0134] The present disclosure provides antibodies or antigen-binding fragments thereof that specifically bind to ANGPTL4 (e.g., human and cynomolgus monkey ANGPTL4), said antibodies or antigen-binding fragments thereof comprising a VL CDR having an amino acid sequence of any one of the VL CDRs of SEQ ID NOs: 17, 18, 19, 20, 21, and / or 22. In particular, the disclosure provides antibodies or antigen-binding fragments thereof that specifically bind to ANGPTL4 (e.g., human and cynomolgus monkey ANGPTL4), said antibodies or antigen-binding fragments thereof comprising (or alternatively, consisting of) one, two, three or more VL CDRs having an amino acid sequence of any of the VL CDRs of SEQ ID NOs: 17, 18, 19, 20, 21, and / or 22. The present disclosure also provides antibodies or antigen-binding fragments thereof that specifically bind to ANGPTL4, said antibodies or antigen-binding fragments thereof comprising a VL domain having an amino acid sequence of SEQ ID NOs: 23 and / or 15.

[0135] Other antibodies or antigen-binding fragments thereof of the disclosure include amino acids that have been mutated, yet have at least 60%, 70%, 80%, 85%, 90% or 95% identity in the CDR regions with the CDR regions depicted in the amino acid sequences described in Table 1. In some embodiments, it includes mutant amino acid sequences wherein no more than 1, 2, 3, 4 or 5 amino acids have been mutated in the CDR regions when compared with the CDR regions depicted in the amino acid sequences described in Table 1.

[0136] The present disclosure also provides nucleic acid sequences that encode VH, VL, full-length heavy chain, and full-length light chain of antibodies or antigen-binding fragments thereof described herein that specifically bind to ANGPTL4 (e.g., human ANGPTL4). Such nucleic acid sequences can be optimized for expression in mammalian cells (for example, Table 1 shows the optimized nucleic acid sequences for heavy chain and light chain of antibodies or antigen-binding fragments thereof of the disclosure).Table 1. Examples of ANGPTL4 Antibodies, Antigen-Binding Fragments and ANGPTL4 Proteins

[0137] Other antibodies or antigen-binding fragments thereof of the disclosure include those where the amino acids or nucleic acids encoding the amino acids have been mutated, yet have at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% identity to the sequences described in Table 1. Some embodiments include amino acid sequences wherein no more than 1, 2, 3, 4 or 5 amino acids have been mutated in the variable regions when compared with the variable regions depicted in the amino acid sequences described in Table 1, while retaining substantially the same antigen binding activity.

[0138] Since each of these antibodies or antigen-binding fragments thereof described herein can bind to ANGPTL4, the VH, VL, full-length light chain, and full-length heavy chain sequences (amino acid sequences and the nucleotide sequences encoding the amino acid sequences) can be “mixed and matched” to create other ANGPTL4-binding antibodies or antigen-binding fragments thereof of the disclosure. Such “mixed and matched” ANGPTL4-binding antibodies or antigen-binding fragments thereof can be tested using binding assays known in the art (e.g., ELISAs, and other assays (e.g., SET assays) as described in e.g., Example 1). When these chains are mixed and matched, a VH sequence from a particular VH / VL pairing can be replaced with a structurally similar VH sequence. Likewise, a full-length heavy chain sequence from a particular full-length heavy chain / full length light chain pairing can be replaced with a structurally similar full-length heavy chain sequence. Likewise, a VL sequence from a particular VH / VL pairing can be replaced with a structurally similar VL sequence. Likewise, a full-length light chain sequence from a particular full-length heavy chain / full-length light chain pairing can be replaced with a structurally similar full-length light chain sequence.

[0139] Accordingly, in one aspect, the disclosure provides an antibody or antigen binding fragment thereof having: a heavy chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 13, 38, 58, 78, 98, 118, and 138, and a light chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23, 48, 68, 88, 108, 128, and 148, wherein the antibody specifically binds to ANGPTL4 (e.g., human ANGPTL4).

[0140] More specifically, in certain aspects, the disclosure provides an antibody or antigen binding fragment thereof having a heavy chain variable domain and a light chain variable domain comprising amino acid sequences selected from SEQ ID NOs: 13 and 23; 38 and 48; 58 and 68; 78 and 88; 98 and 108, 118 and 128, or 138 and 148, respectively.

[0141] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain variable domain and a light chain variable domain comprising amino acid sequences found in Table 1 for use as a medicament.

[0142] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain variable domain and a light chain variable domain comprising amino acid sequences found in Table 1 for use in the treatment of a disorder.

[0143] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain variable domain and a light chain variable domain comprising amino acid sequences found in Table 1 for the manufacture of a medicament for the treatment of a disorder.

[0144] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain variable domain and a light chain variable domain comprising amino acid sequences selected from SEQ ID NOs: 13 and 23 for use as a medicament.

[0145] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain variable domain and a light chain variable domain comprising amino acid sequences selected from SEQ ID NOs: 13 and 23 for use in the treatment of a disorder.

[0146] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chainvariable domain and a light chain variable domain comprising amino acid sequences selected from SEQ ID NOs: 13 and 23 for the manufacture of a medicament for the treatment of a disorder.

[0147] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain variable domain and a light chain variable domain comprising amino acid sequences selected from SEQ ID NOs: 38 and 48 for use as a medicament.

[0148] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain variable domain and a light chain variable domain comprising amino acid sequences selected from SEQ ID NOs: 38 and 48 for use in the treatment of a disorder.

[0149] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain variable domain and a light chain variable domain comprising amino acid sequences selected from SEQ ID NOs: 38 and 48 for the manufacture of a medicament for the treatment of a disorder.

[0150] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain variable domain and a light chain variable domain comprising amino acid sequences selected from SEQ ID NOs: 58 and 68 for use as a medicament.

[0151] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain variable domain and a light chain variable domain comprising amino acid sequences selected from SEQ ID NOs: 58 and 68 for use in the treatment of a disorder.

[0152] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain variable domain and a light chain variable domain comprising amino acid sequences selected from SEQ ID NOs: 58 and 68 for the manufacture of a medicament for the treatment of a disorder.

[0153] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain variabledomain and a light chain variable domain comprising amino acid sequences selected from SEQ ID NOs: 78 and 88 for use as a medicament.

[0154] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain variable domain and a light chain variable domain comprising amino acid sequences selected from SEQ ID NOs: 78 and 88 for use in the treatment of a disorder.

[0155] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain variable domain and a light chain variable domain comprising amino acid sequences selected from SEQ ID NOs: 78 and 88 for the manufacture of a medicament for the treatment of a disorder.

[0156] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain variable domain and a light chain variable domain comprising amino acid sequences selected from SEQ ID NOs: 98 and 108 for use as a medicament.

[0157] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain variable domain and a light chain variable domain comprising amino acid sequences selected from SEQ ID NOs: 98 and 108 for use in the treatment of a disorder.

[0158] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain variable domain and a light chain variable domain comprising amino acid sequences selected from SEQ ID NOs: 98 and 108 for the manufacture of a medicament for the treatment of a disorder.

[0159] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain variable domain and a light chain variable domain comprising amino acid sequences selected from SEQ ID Nos: 118 and 128 for use as a medicament.

[0160] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain variable domain and a light chain variable domain comprising amino acid sequences selected from SEQ ID Nos: 118 and 128 for use in the treatment of a disorder.

[0161] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain variable domain and a light chain variable domain comprising amino acid sequences selected from SEQ ID NOs: 118 and 128 for the manufacture of a medicament for the treatment of a disorder.

[0162] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain variable domain and a light chain variable domain comprising amino acid sequences selected from SEQ ID Nos: 138 and 148 for use as a medicament.

[0163] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain variable domain and a light chain variable domain comprising amino acid sequences selected from SEQ ID Nos: 138 and 148 for use in the treatment of a disorder.

[0164] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain variable domain and a light chain variable domain comprising amino acid sequences selected from SEQ ID NOs: 138 and 148 for the manufacture of a medicament for the treatment of a disorder.

[0165] In another aspect, the disclosure provides (i) an antibody having: a full-length heavy chain comprising an amino acid sequence that has been optimized for expression in a mammalian cell selected from the group consisting of SEQ ID NOs: 15, 28, 40, 60, 80, 100, 120, 140, 156, and 157, and a full-length light chain comprising an amino acid sequence that has been optimized for expression in a mammalian cell selected from the group consisting of SEQ ID NOs: 25, 50, 70, 90, 110, 130, and 150; or (ii) a functional protein comprising an antigen binding portion thereof. More specifically, in certain aspects, the disclosure provides an antibody or antigen binding fragment thereof having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 15 and 25; 28 and 25; 40 and 50; 60 and 70; 80 and 90; 100 and 110; 120 and 130; or 140 and 150, respectively.

[0166] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences found in Table 1 for use as a medicament.

[0167] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences found in Table 1 for use in the treatment of a disorder.

[0168] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences found in Table 1 for the manufacture of a medicament for the treatment of a disorder.

[0169] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 15 and 25 for use as a medicament.

[0170] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 15 and 25 for use in the treatment of a disorder.

[0171] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 15 and 25 for the manufacture of a medicament for the treatment of a disorder.

[0172] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 28 and 25 for use as a medicament.

[0173] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 28 and 25 for use in the treatment of a disorder.

[0174] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 28 and 25 for the manufacture of a medicament for the treatment of a disorder.

[0175] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 156 and 25 for use as a medicament.

[0176] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 156 and 25 for use in the treatment of a disorder.

[0177] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 156 and 25 for the manufacture of a medicament for the treatment of a disorder.

[0178] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 157 and 25 for use as a medicament.

[0179] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 157 and 25 for use in the treatment of a disorder.

[0180] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 157 and 25 for the manufacture of a medicament for the treatment of a disorder.

[0181] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 40 and 50 for use as a medicament.

[0182] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 40 and 50 for use in the treatment of a disorder.

[0183] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 40 and 50 for the manufacture of a medicament for the treatment of a disorder.

[0184] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 60 and 70 for use as a medicament.

[0185] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 60 and 70 for use in the treatment of a disorder.

[0186] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 60 and 70 for the manufacture of a medicament for the treatment of a disorder.

[0187] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 80 and 90 for use as a medicament.

[0188] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 80 and 90 for use in the treatment of a disorder.

[0189] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 80 and 90 for the manufacture of a medicament for the treatment of a disorder.

[0190] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 100 and 110 for use as a medicament.

[0191] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 100 and 110 for use in the treatment of a disorder.

[0192] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 100 and 110 for the manufacture of a medicament for the treatment of a disorder.

[0193] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 120 and 130 for use as a medicament.

[0194] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 120 and 130 for use in the treatment of a disorder.

[0195] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 120 and 130 for the manufacture of a medicament for the treatment of a disorder.

[0196] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 140 and 150 for use as a medicament.

[0197] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 140 and 150 for use in the treatment of a disorder.

[0198] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and a light chain comprising amino acid sequences selected from SEQ ID NOs: 140 and 150 for the manufacture of a medicament for the treatment of a disorder.

[0199] In another aspect, the present disclosure provides ANGPTL4 binding antibodies or antigen-binding fragments thereof that comprise the heavy chain and light chain CDRls, CDR2s, and CDR3s as described in Table 1, or combinations thereof.

[0200] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and light chain CDRls, CDR2s, and CDR3s as described in Table 1, or combinations thereof for use as a medicament.

[0201] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and light chain CDRls, CDR2s, and CDR3s as described in Table 1, or combinations thereof for use in the treatment of a disorder.

[0202] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 having a heavy chain and light chain CDRls, CDR2s, and CDR3s as described in Table 1, or combinations thereof for the manufacture of a medicament for the treatment of a disorder for the manufacture of a medicament for the treatment of a disorder .

[0203] In some embodiments, the amino acid sequences of the VH CDRls of the antibodies or antigen-binding fragments thereof are shown in SEQ ID NOs: 7, 32, 52, 72, 92, 112, and 132. In some embodiments, the amino acid sequences of the VH CDR2s of the antibodies or antigen-binding fragments thereof and are shown in SEQ ID NOs: 8, 33, 53,73, 93, 113, and 133. In some embodiments, the amino acid sequences of the VH CDR3s of the antibodies or antigen-binding fragments thereof are shown in SEQ ID NOs: 9, 34, 54,74, 94, 114, and 134. In some embodiments, the amino acid sequences of the VL CDRls of the antibodies or antigen -binding fragments thereof are shown in SEQ ID NOs: 17, 42, 62, 82, 102, 122, and 142. In some embodiments, the amino acid sequences of the VL CDR2s of the antibodies or antigen-binding fragments thereof are shown in SEQ ID NOs: 18, 43, 63, 83, 103, 123, and 143. In some embodiments, the amino acid sequences of the VL CDR3s of the antibodies or antigen-binding fragments thereof are shown in SEQ ID NOs: 19, 44, 64, 84, 104, 124, and 144. These CDR regions are delineated using the Kabat system.

[0204] Alternatively, as defined using the Chothia system (Al-Lazikani et al., (1997) JMB 273,927-948), the amino acid sequences of the VH CDRls of the antibodies orantigen-binding fragments thereof are shown in SEQ ID NOs: 10, 35, 55, 75, 95, 115, and135. In some embodiments, the amino acid sequences of the VH CDR2s of the antibodies or antigen-binding fragments thereof are shown in SEQ ID NOs: 11, 36, 56, 76, 96, 116, and136. In some embodiments, the amino acid sequences of the VH CDR3s of the antibodies or antigen-binding fragments thereof are shown in SEQ ID NOs: 12, 37, 57, 77, 97, 117, 117, and 137. In some embodiments, the amino acid sequences of the VL CDRls of the antibodies or antigen-binding fragments thereof are shown in SEQ ID NOs: 20, 45, 65, 85, 105, 125, and 145. In some embodiments, the amino acid sequences of the VL CDR2s of the antibodies or antigen-binding fragments thereof are shown in SEQ ID NOs: 21, 46, 66, 86, 106, 126, and 146. In some embodiments, the amino acid sequences of the VL CDR3s of the antibodies or antigen-binding fragments thereof are shown in SEQ ID NOs: 22, 47, 67, 87, 107, 127, and 147.

[0205] In some embodiments, the disclosure includes an antibody or antigen-binding fragment thereof that specifically binds to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 7; a heavy chain variable region CDR2 of SEQ ID NO: 8; a heavy chain variable region CDR3 of SEQ ID NO: 9; a light chain variable region CDR1 of SEQ ID NO: 17; a light chain variable region CDR2 of SEQ ID NO: 18; and a light chain variable region CDR3 of SEQ ID NO: 19.

[0206] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 7; a heavy chain variable region CDR2 of SEQ ID NO: 8; a heavy chain variable region CDR3 of SEQ ID NO: 9; a light chain variable region CDR1 of SEQ ID NO: 17; a light chain variable region CDR2 of SEQ ID NO: 18; and a light chain variable region CDR3 of SEQ ID NO: 19 for use as a medicament.

[0207] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 7; a heavy chain variable region CDR2 of SEQ ID NO: 8; a heavy chain variable region CDR3 of SEQ ID NO: 9; a light chain variable region CDR1 of SEQ ID NO: 17; a light chain variable region CDR2 of SEQ ID NO: 18; and a light chain variable region CDR3 of SEQ ID NO: 19 for use in the treatment of a disorder.

[0208] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chainvariable region CDR1 of SEQ ID NO: 7; a heavy chain variable region CDR2 of SEQ ID NO: 8; a heavy chain variable region CDR3 of SEQ ID NO: 9; a light chain variable region CDR1 of SEQ ID NO: 17; a light chain variable region CDR2 of SEQ ID NO: 18; and a light chain variable region CDR3 of SEQ ID NO: 19 for the manufacture of a medicament for the treatment of a disorder.

[0209] In some embodiments, the disclosure includes an antibody or antigen-binding fragment thereof that specifically binds to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 32; a heavy chain variable region CDR2 of SEQ ID NO: 33; a heavy chain variable region CDR3 of SEQ ID NO: 34; a light chain variable region CDR1 of SEQ ID NO: 42; a light chain variable region CDR2 of SEQ ID NO: 43; and a light chain variable region CDR3 of SEQ ID NO: 44.

[0210] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 32; a heavy chain variable region CDR2 of SEQ ID NO: 33; a heavy chain variable region CDR3 of SEQ ID NO: 34; a light chain variable region CDR1 of SEQ ID NO: 42; a light chain variable region CDR2 of SEQ ID NO: 43; and a light chain variable region CDR3 of SEQ ID NO: 44 for use as a medicament.

[0211] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 32; a heavy chain variable region CDR2 of SEQ ID NO: 33; a heavy chain variable region CDR3 of SEQ ID NO: 34; a light chain variable region CDR1 of SEQ ID NO: 42; a light chain variable region CDR2 of SEQ ID NO: 43; and a light chain variable region CDR3 of SEQ ID NO: 44 for use in the treatment of a disorder.

[0212] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 32; a heavy chain variable region CDR2 of SEQ ID NO: 33; a heavy chain variable region CDR3 of SEQ ID NO: 34; a light chain variable region CDR1 of SEQ ID NO: 42; a light chain variable region CDR2 of SEQ ID NO: 43; and a light chain variable region CDR3 of SEQ ID NO: 44 for the manufacture of a medicament for the treatment of a disorder.

[0213] In some embodiments, the disclosure includes an antibody or antigen-binding fragment thereof that specifically binds to ANGPTL4 comprising a heavy chain variableregion CDR1 of SEQ ID NO: 52; a heavy chain variable region CDR2 of SEQ ID NO: 53; a heavy chain variable region CDR3 of SEQ ID NO: 54; a light chain variable region CDR1 of SEQ ID NO: 62; a light chain variable region CDR2 of SEQ ID NO: 63; and a light chain variable region CDR3 of SEQ ID NO: 64.

[0214] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 52; a heavy chain variable region CDR2 of SEQ ID NO: 53; a heavy chain variable region CDR3 of SEQ ID NO: 54; a light chain variable region CDR1 of SEQ ID NO: 62; a light chain variable region CDR2 of SEQ ID NO: 63; and a light chain variable region CDR3 of SEQ ID NO: 64 for use as a medicament.

[0215] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 52; a heavy chain variable region CDR2 of SEQ ID NO: 53; a heavy chain variable region CDR3 of SEQ ID NO: 54; a light chain variable region CDR1 of SEQ ID NO: 62; a light chain variable region CDR2 of SEQ ID NO: 63; and a light chain variable region CDR3 of SEQ ID NO: 64 for use in the treatment of a disorder.

[0216] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 52; a heavy chain variable region CDR2 of SEQ ID NO: 53; a heavy chain variable region CDR3 of SEQ ID NO: 54; a light chain variable region CDR1 of SEQ ID NO: 62; a light chain variable region CDR2 of SEQ ID NO: 63; and a light chain variable region CDR3 of SEQ ID NO: 64 for the manufacture of a medicament for the treatment of a disorder.

[0217] In some embodiments, the disclosure includes an antibody or antigen-binding fragment thereof that specifically binds to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 72; a heavy chain variable region CDR2 of SEQ ID NO: 73; a heavy chain variable region CDR3 of SEQ ID NO: 74; a light chain variable region CDR1 of SEQ ID NO: 82; a light chain variable region CDR2 of SEQ ID NO: 83; and a light chain variable region CDR3 of SEQ ID NO: 84.

[0218] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 72; a heavy chain variable region CDR2 of SEQ ID NO: 73; aheavy chain variable region CDR3 of SEQ ID NO: 74; a light chain variable region CDR1 of SEQ ID NO: 82; a light chain variable region CDR2 of SEQ ID NO: 83; and a light chain variable region CDR3 of SEQ ID NO: 84 for use as a medicament.

[0219] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 72; a heavy chain variable region CDR2 of SEQ ID NO: 73; a heavy chain variable region CDR3 of SEQ ID NO: 74; a light chain variable region CDR1 of SEQ ID NO: 82; a light chain variable region CDR2 of SEQ ID NO: 83; and a light chain variable region CDR3 of SEQ ID NO: 84 for use in the treatment of a disorder.

[0220] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 72; a heavy chain variable region CDR2 of SEQ ID NO: 73; a heavy chain variable region CDR3 of SEQ ID NO: 74; a light chain variable region CDR1 of SEQ ID NO: 82; a light chain variable region CDR2 of SEQ ID NO: 83; and a light chain variable region CDR3 of SEQ ID NO: 84 for the manufacture of a medicament for the treatment of a disorder.

[0221] In some embodiments, the disclosure includes an antibody or antigen-binding fragment thereof that specifically binds to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 92; a heavy chain variable region CDR2 of SEQ ID NO: 93; a heavy chain variable region CDR3 of SEQ ID NO: 94; a light chain variable region CDR1 of SEQ ID NO: 102; a light chain variable region CDR2 of SEQ ID NO: 103; and a light chain variable region CDR3 of SEQ ID NO: 104.

[0222] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 92; a heavy chain variable region CDR2 of SEQ ID NO: 93; a heavy chain variable region CDR3 of SEQ ID NO: 94; a light chain variable region CDR1 of SEQ ID NO: 102; a light chain variable region CDR2 of SEQ ID NO: 103; and a light chain variable region CDR3 of SEQ ID NO: 104 for use as a medicament.

[0223] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 92; a heavy chain variable region CDR2 of SEQ ID NO: 93; a heavy chain variable region CDR3 of SEQ ID NO: 94; a light chain variable region CDR1of SEQ ID NO: 102; a light chain variable region CDR2 of SEQ ID NO: 103; and a light chain variable region CDR3 of SEQ ID NO: 104 for use in the treatment of a disorder.

[0224] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 92; a heavy chain variable region CDR2 of SEQ ID NO: 93; a heavy chain variable region CDR3 of SEQ ID NO: 94; a light chain variable region CDR1 of SEQ ID NO: 102; a light chain variable region CDR2 of SEQ ID NO: 103; and a light chain variable region CDR3 of SEQ ID NO: 104 for the manufacture of a medicament for the treatment of a disorder.

[0225] In some embodiments, the disclosure includes an antibody or antigen-binding fragment thereof that specifically binds to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 112; a heavy chain variable region CDR2 of SEQ ID NO: 113; a heavy chain variable region CDR3 of SEQ ID NO: 114; a light chain variable region CDR1 of SEQ ID NO: 122; a light chain variable region CDR2 of SEQ ID NO: 123; and a light chain variable region CDR3 of SEQ ID NO: 124.

[0226] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 112; a heavy chain variable region CDR2 of SEQ ID NO: 113; a heavy chain variable region CDR3 of SEQ ID NO: 114; a light chain variable region CDR1 of SEQ ID NO: 122; a light chain variable region CDR2 of SEQ ID NO: 123; and a light chain variable region CDR3 of SEQ ID NO: 124 for use as a medicament.

[0227] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 112; a heavy chain variable region CDR2 of SEQ ID NO: 113; a heavy chain variable region CDR3 of SEQ ID NO: 114; a light chain variable region CDR1 of SEQ ID NO: 122; a light chain variable region CDR2 of SEQ ID NO: 123; and a light chain variable region CDR3 of SEQ ID NO: 124 for use in the treatment of a disorder.

[0228] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 112; a heavy chain variable region CDR2 of SEQ ID NO: 113; a heavy chain variable region CDR3 of SEQ ID NO: 114; a light chain variable region CDR1 of SEQ ID NO: 122; a light chain variable region CDR2 of SEQ ID NO: 123;and a light chain variable region CDR3 of SEQ ID NO: 124 for the manufacture of a medicament for the treatment of a disorder.

[0229] In some embodiments, the disclosure includes an antibody or antigen-binding fragment thereof that specifically binds to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 132; a heavy chain variable region CDR2 of SEQ ID NO: 133; a heavy chain variable region CDR3 of SEQ ID NO: 134; a light chain variable region CDR1 of SEQ ID NO: 142; a light chain variable region CDR2 of SEQ ID NO: 143; and a light chain variable region CDR3 of SEQ ID NO: 144.

[0230] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 132; a heavy chain variable region CDR2 of SEQ ID NO: 133; a heavy chain variable region CDR3 of SEQ ID NO: 134; a light chain variable region CDR1 of SEQ ID NO: 142; a light chain variable region CDR2 of SEQ ID NO: 143; and a light chain variable region CDR3 of SEQ ID NO: 144 for use as a medicament.

[0231] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 132; a heavy chain variable region CDR2 of SEQ ID NO: 133; a heavy chain variable region CDR3 of SEQ ID NO: 134; a light chain variable region CDR1 of SEQ ID NO: 142; a light chain variable region CDR2 of SEQ ID NO: 143; and a light chain variable region CDR3 of SEQ ID NO: 144 for use in the treatment of a disorder.

[0232] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 132; a heavy chain variable region CDR2 of SEQ ID NO: 133; a heavy chain variable region CDR3 of SEQ ID NO: 134; a light chain variable region CDR1 of SEQ ID NO: 142; a light chain variable region CDR2 of SEQ ID NO: 143; and a light chain variable region CDR3 of SEQ ID NO: 144 for the manufacture of a medicament for the treatment of a disorder.

[0233] In some embodiments, the disclosure includes an antibody or antigen-binding fragment thereof that specifically binds to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 10; a heavy chain variable region CDR2 of SEQ ID NO: 11; a heavy chain variable region CDR3 of SEQ ID NO: 12; a light chain variable region CDR1of SEQ ID NO: 20; a light chain variable region CDR2 of SEQ ID NO: 21; and a light chain variable region CDR3 of SEQ ID NO: 22.

[0234] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 10; a heavy chain variable region CDR2 of SEQ ID NO: 11; a heavy chain variable region CDR3 of SEQ ID NO: 12; a light chain variable region CDR1 of SEQ ID NO: 20; a light chain variable region CDR2 of SEQ ID NO: 21; and a light chain variable region CDR3 of SEQ ID NO: 22 for use as a medicament.

[0235] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 10; a heavy chain variable region CDR2 of SEQ ID NO: 11; a heavy chain variable region CDR3 of SEQ ID NO: 12; a light chain variable region CDR1 of SEQ ID NO: 20; a light chain variable region CDR2 of SEQ ID NO: 21; and a light chain variable region CDR3 of SEQ ID NO: 22 for use in the treatment of a disorder.

[0236] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 10; a heavy chain variable region CDR2 of SEQ ID NO: 11; a heavy chain variable region CDR3 of SEQ ID NO: 12; a light chain variable region CDR1 of SEQ ID NO: 20; a light chain variable region CDR2 of SEQ ID NO: 21; and a light chain variable region CDR3 of SEQ ID NO: 22 for the manufacture of a medicament for the treatment of a disorder.

[0237] In some embodiments, the disclosure includes an antibody or antigen-binding fragment thereof that specifically binds to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 35; a heavy chain variable region CDR2 of SEQ ID NO: 36; a heavy chain variable region CDR3 of SEQ ID NO: 37; a light chain variable region CDR1 of SEQ ID NO: 45; a light chain variable region CDR2 of SEQ ID NO: 46; and a light chain variable region CDR3 of SEQ ID NO: 47.

[0238] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 35; a heavy chain variable region CDR2 of SEQ ID NO: 36; a heavy chain variable region CDR3 of SEQ ID NO: 37; a light chain variable region CDR1of SEQ ID NO: 45; a light chain variable region CDR2 of SEQ ID NO: 46; and a light chain variable region CDR3 of SEQ ID NO: 47 for use as a medicament.

[0239] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 35; a heavy chain variable region CDR2 of SEQ ID NO: 36; a heavy chain variable region CDR3 of SEQ ID NO: 37; a light chain variable region CDR1 of SEQ ID NO: 45; a light chain variable region CDR2 of SEQ ID NO: 46; and a light chain variable region CDR3 of SEQ ID NO: 47 for use in the treatment of a disorder.

[0240] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 35; a heavy chain variable region CDR2 of SEQ ID NO: 36; a heavy chain variable region CDR3 of SEQ ID NO: 37; a light chain variable region CDR1 of SEQ ID NO: 45; a light chain variable region CDR2 of SEQ ID NO: 46; and a light chain variable region CDR3 of SEQ ID NO: 47 for the manufacture of a medicament for the treatment of a disorder.

[0241] In some embodiments, the disclosure includes an antibody or antigen-binding fragment thereof that specifically binds to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 55; a heavy chain variable region CDR2 of SEQ ID NO: 56; a heavy chain variable region CDR3 of SEQ ID NO: 57; a light chain variable region CDR1 of SEQ ID NO: 65; a light chain variable region CDR2 of SEQ ID NO: 66; and a light chain variable region CDR3 of SEQ ID NO: 67.

[0242] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 55; a heavy chain variable region CDR2 of SEQ ID NO: 56; a heavy chain variable region CDR3 of SEQ ID NO: 57; a light chain variable region CDR1 of SEQ ID NO: 65; a light chain variable region CDR2 of SEQ ID NO: 66; and a light chain variable region CDR3 of SEQ ID NO: 67 for use as a medicament.

[0243] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 55; a heavy chain variable region CDR2 of SEQ ID NO: 56; a heavy chain variable region CDR3 of SEQ ID NO: 57; a light chain variable region CDR1of SEQ ID NO: 65; a light chain variable region CDR2 of SEQ ID NO: 66; and a light chain variable region CDR3 of SEQ ID NO: 67 for use in the treatment of a disorder.

[0244] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 55; a heavy chain variable region CDR2 of SEQ ID NO: 56; a heavy chain variable region CDR3 of SEQ ID NO: 57; a light chain variable region CDR1 of SEQ ID NO: 65; a light chain variable region CDR2 of SEQ ID NO: 66; and a light chain variable region CDR3 of SEQ ID NO: 67 for the manufacture of a medicament for the treatment of a disorder.

[0245] In some embodiments, the disclosure includes an antibody or antigen-binding fragment thereof that specifically binds to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 75; a heavy chain variable region CDR2 of SEQ ID NO: 76; a heavy chain variable region CDR3 of SEQ ID NO: 77; a light chain variable region CDR1 of SEQ ID NO: 85; a light chain variable region CDR2 of SEQ ID NO: 86; and a light chain variable region CDR3 of SEQ ID NO: 87.

[0246] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 75; a heavy chain variable region CDR2 of SEQ ID NO: 76; a heavy chain variable region CDR3 of SEQ ID NO: 77; a light chain variable region CDR1 of SEQ ID NO: 85; a light chain variable region CDR2 of SEQ ID NO: 86; and a light chain variable region CDR3 of SEQ ID NO: 87 for use as a medicament.

[0247] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 75; a heavy chain variable region CDR2 of SEQ ID NO: 76; a heavy chain variable region CDR3 of SEQ ID NO: 77; a light chain variable region CDR1 of SEQ ID NO: 85; a light chain variable region CDR2 of SEQ ID NO: 86; and a light chain variable region CDR3 of SEQ ID NO: 87 for use in the treatment of a disorder.

[0248] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 75; a heavy chain variable region CDR2 of SEQ ID NO: 76; a heavy chain variable region CDR3 of SEQ ID NO: 77; a light chain variable region CDR1 of SEQ ID NO: 85; a light chain variable region CDR2 of SEQ ID NO: 86;and a light chain variable region CDR3 of SEQ ID NO: 87 for the manufacture of a medicament for the treatment of a disorder. In some embodiments, the disclosure includes an antibody or antigen-binding fragment thereof that specifically binds to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 95; a heavy chain variable region CDR2 of SEQ ID NO: 96; a heavy chain variable region CDR3 of SEQ ID NO: 97; a light chain variable region CDR1 of SEQ ID NO: 105; a light chain variable region CDR2 of SEQ ID NO: 106; and a light chain variable region CDR3 of SEQ ID NO: 107.

[0249] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 95; a heavy chain variable region CDR2 of SEQ ID NO: 96; a heavy chain variable region CDR3 of SEQ ID NO: 97; a light chain variable region CDR1 of SEQ ID NO: 105; a light chain variable region CDR2 of SEQ ID NO: 106; and a light chain variable region CDR3 of SEQ ID NO: 107 for use as a medicament.

[0250] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 95; a heavy chain variable region CDR2 of SEQ ID NO: 96; a heavy chain variable region CDR3 of SEQ ID NO: 97; a light chain variable region CDR1 of SEQ ID NO: 105; a light chain variable region CDR2 of SEQ ID NO: 106; and a light chain variable region CDR3 of SEQ ID NO: 107 for use in the treatment of a disorder.

[0251] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 95; a heavy chain variable region CDR2 of SEQ ID NO: 96; a heavy chain variable region CDR3 of SEQ ID NO: 97; a light chain variable region CDR1 of SEQ ID NO: 105; a light chain variable region CDR2 of SEQ ID NO: 106; and a light chain variable region CDR3 of SEQ ID NO: 107 for the manufacture of a medicament for the treatment of a disorder.

[0252] In some embodiments, the disclosure includes an antibody or antigen-binding fragment thereof that specifically binds to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 115; a heavy chain variable region CDR2 of SEQ ID NO: 116; a heavy chain variable region CDR3 of SEQ ID NO: 117; a light chain variable region CDR1 of SEQ ID NO: 125; a light chain variable region CDR2 of SEQ ID NO: 126; and a light chain variable region CDR3 of SEQ ID NO: 127.

[0253] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 115; a heavy chain variable region CDR2 of SEQ ID NO: 116; a heavy chain variable region CDR3 of SEQ ID NO: 117; a light chain variable region CDR1 of SEQ ID NO: 125; a light chain variable region CDR2 of SEQ ID NO: 126; and a light chain variable region CDR3 of SEQ ID NO: 127 for use as a medicament.

[0254] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 115; a heavy chain variable region CDR2 of SEQ ID NO: 116; a heavy chain variable region CDR3 of SEQ ID NO: 117; a light chain variable region CDR1 of SEQ ID NO: 125; a light chain variable region CDR2 of SEQ ID NO: 126; and a light chain variable region CDR3 of SEQ ID NO: 127 for use in the treatment of a disorder.

[0255] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 115; a heavy chain variable region CDR2 of SEQ ID NO: 116; a heavy chain variable region CDR3 of SEQ ID NO: 117; a light chain variable region CDR1 of SEQ ID NO: 125; a light chain variable region CDR2 of SEQ ID NO: 126; and a light chain variable region CDR3 of SEQ ID NO: 127 for the manufacture of a medicament for the treatment of a disorder.

[0256] In some embodiments, the disclosure includes an antibody or antigen-binding fragment thereof that specifically binds to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 135; a heavy chain variable region CDR2 of SEQ ID NO: 136; a heavy chain variable region CDR3 of SEQ ID NO: 137; a light chain variable region CDR1 of SEQ ID NO: 145; a light chain variable region CDR2 of SEQ ID NO: 146; and a light chain variable region CDR3 of SEQ ID NO: 147.

[0257] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 135; a heavy chain variable region CDR2 of SEQ ID NO: 136; a heavy chain variable region CDR3 of SEQ ID NO: 137; a light chain variable region CDR1 of SEQ ID NO: 145; a light chain variable region CDR2 of SEQ ID NO: 146; and a light chain variable region CDR3 of SEQ ID NO: 147 for use as a medicament.

[0258] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 135; a heavy chain variable region CDR2 of SEQ ID NO: 136; a heavy chain variable region CDR3 of SEQ ID NO: 137; a light chain variable region CDR1 of SEQ ID NO: 145; a light chain variable region CDR2 of SEQ ID NO: 146; and a light chain variable region CDR3 of SEQ ID NO: 147 for use in the treatment of a disorder.

[0259] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to ANGPTL4 comprising a heavy chain variable region CDR1 of SEQ ID NO: 135; a heavy chain variable region CDR2 of SEQ ID NO: 136; a heavy chain variable region CDR3 of SEQ ID NO: 137; a light chain variable region CDR1 of SEQ ID NO: 145; a light chain variable region CDR2 of SEQ ID NO: 146; and a light chain variable region CDR3 of SEQ ID NO: 147 for the manufacture of a medicament for the treatment of a disorder.

[0260] In some embodiments, the disclosure includes antibodies or antigen binding fragments that specifically bind to ANGPTL4 as described in Table 1.

[0261] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 as described in Table 1 for use as a medicament.

[0262] In some embodiments, the disclosure provides an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 as described in Table 1 for use in the treatment of a disorder.

[0263] In some embodiments, the disclosure provides use of an antibody or antigen binding fragment thereof that binds specifically to human ANGPTL4 as described in Table 1, or combinations thereof for the manufacture of a medicament for the treatment of a disorder.

[0264] In a preferred embodiment, the antibody, or antigen-binding fragment, that binds ANGPTL4 is NEG276, NEG276-LALA, NEG278, NEG310, NEG313, NEG315, NEG318, and NEG319.

[0265] The ANGPTL4 binding antibodies, or antigen-binding fragments thereof, as described herein can be monoclonal antibodies, human or humanized antibodies, chimeric antibodies, single chain antibodies, Fab fragments, Fv fragments, F(ab’)2 fragments, or scFv fragments, and / or IgG isotypes (e.g., IgM, IgE, IgG such as IgGl or IgG4). In someembodiments, ANGPTL4 binding antibodies or antigen-binding fragments thereof is an IgGl or IgG4 isotype.Antibodies with extended half life

[0266] The present disclosure provides antibodies or antigen-binding fragments thereof that bind specifically to ANGPTL4 which have an extended half-life in vivo. Many factors may affect a protein’s half-life in vivo. For examples, kidney filtration, metabolism in the liver, degradation by proteolytic enzymes (proteases), and immunogenic responses (e.g., protein neutralization by antibodies and uptake by macrophages and dendritic cells). A variety of strategies can be used to extend the half-life of antibodies of the present disclosure. For example, by chemical linkage to polyethyleneglycol (PEG), reCODE PEG, antibody scaffold, polysialic acid (PSA), hydroxy ethyl starch (HES), albumin-binding ligands, and carbohydrate shields; by genetic fusion to proteins binding to serum proteins, such as albumin, IgG, FcRn, and transferring; by coupling (genetically or chemically) to other binding moieties that bind to serum proteins, such as nanobodies, Fabs, DARPins, avimers, affibodies, and anticalins; by genetic fusion to rPEG, albumin, domain of albumin, albumin-binding proteins, and Fc; or by incorporation into nanocarriers, slow release formulations, or medical devices.

[0267] To prolong the serum circulation of antibodies in vivo, inert polymer molecules such as high molecular weight PEG can be attached to the antibodies or antigenbinding fragments thereof with or without a multifunctional linker either through sitespecific conjugation of the PEG to the N- or C-terminus of the antibodies or via epsilon- amino groups present on lysine residues. To pegylate an antibody or antigen-binding fragment thereof, an antibody or antigen-binding fragment thereof typically is reacted with PEG, such as a reactive ester or aldehyde derivative of PEG, under conditions in which one or more PEG groups become attached to the antibody or antibody fragment. The pegylation can be carried out by an acylation reaction or an alkylation reaction with a reactive PEG molecule (or an analogous reactive water-soluble polymer). As used herein, the term “polyethylene glycol” is intended to encompass any of the forms of PEG that have been used to derivatize other proteins, such as mono (Cl -CIO) alkoxy- or aryloxy-polyethylene glycol or polyethylene glycol-maleimide. In certain embodiments, an antibody or antigen-binding fragment thereof to be pegylated is an aglycosylated antibody or antigen-binding fragment thereof. Linear or branched polymer derivatization that results in minimal loss of biological activity will be used. The degree of conjugation can be closely monitored by SDS-PAGE and mass spectrometry to ensure proper conjugation of PEG molecules to an antibody or antigen-binding fragment thereof. Unreacted PEG can be separated from antibody-PEG conjugates by size-exclusion or by ion-exchange chromatography. PEG- derivatized antibodies or antigen-binding fragments thereof can be tested for binding activity as well as for in vivo efficacy using methods well-known to those of skill in the art, for example, by immunoassays. Methods for pegylating proteins are known in the art and can be applied to the antibodies or antigen-binding fragments thereof of the present disclosure.

[0268] Another technology includes the use of hydroxy ethyl starch (“HES”) derivatives linked to antibodies or antigen-binding fragments thereof. HES is a modified natural polymer derived from waxy maize starch and can be metabolized by the body’s enzymes. HES solutions are usually administered to substitute deficient blood volume and to improve the rheological properties of the blood. Hesylation of an antibody or antigenbinding fragments thereof enables the prolongation of the circulation half-life by increasing the stability of the molecule, as well as by reducing renal clearance, resulting in an increased biological activity. By varying different parameters, such as the molecular weight of HES, a wide range of HES antibody conjugates can be customized.

[0269] Antibodies or antigen-binding fragments thereof having an increased half-life can also be generated by introducing one or more amino acid modifications (i.e., substitutions, insertions or deletions) into an IgG constant domain, or FcRn binding fragment thereof. See, e.g., International Publication No. WO 98 / 23289; International Publication No. WO 97 / 34631; and U.S. Patent No. 6,277,375, each of which is hereby incorporated by reference in its entirety. One or more of the following non-limiting examples of glycosylation-reducing amino acid substitutions may be useful in increasing antibody half-life: M252Y, S254T, T256E, N297A, L234A, L235A, C220, C226S, C229S, P238S, C226S, C229S, E323P, L234V, L235A, G237A, L234F, N297G, L235E, P329G, P331S, M428L, N434S, or combinations thereof, numbering according to EU index. Mutations M428L and N434S have been particularly useful in increasing half-life of an antibody and improving its binding affinity to FcRn relative to an unmodified antibody (see, e.g., Zalevsky et al., 2010, Nat Biotechnol. 28(2): 157-159). Mutations M252Y, S254T, andT256E have also been particularly useful in increasing half-life of an antibody and improving its binding affinity to FcRn relative to an unmodified antibody (see, e.g., Dall'Acqua et al., 2006, J Biol Chem. 18;281(33):23514-24. ).

[0270] Further, antibodies or antigen-binding fragments thereof can be conjugated to albumin (e.g., human serum albumin; HSA) to make the antibody or antigen-binding fragment thereof more stable in vivo or have a longer half-life in vivo. The techniques are well-known in the art, see, e.g., International Publication Nos. WO 93 / 15199, WO 93 / 15200, and WO 01 / 77137; and European Patent No. EP 413622, each of which is hereby incorporated by reference in its entirety.Methods of making

[0271] The present disclosure, among other things, provides methods of making ANGPTL4 binding antibodies or antigen-binding fragments thereof described herein. In some embodiments, an ANGPTL4 binding antibody or an antigen-binding fragment thereof described herein is identified using a display technology, such as yeast display, phage display, or ribosome display. In some embodiments, an ANGPTL4 binding antibody or an antigen-binding fragment thereof described herein is identified using a hybridoma library (e.g., a mammalian hybridoma library, e.g., a mouse hybridoma library), followed by supernatant screening.

[0272] Combinatorial methods for generating antibodies or antigen-bindings fragments thereof are known in the art (as described in, e.g., Ladner et al. U.S. Patent No. 5,223,409; Kang et al. International Publication No. WO 92 / 18619; Dower et al. International Publication No. WO 91 / 17271; Winter et al. International Publication WO 92 / 20791; Markland et al. International Publication No. WO 92 / 15679; Breitling et al. International Publication WO 93 / 01288; McCafferty et al. International Publication No. WO 92 / 01047; Garrard et al. International Publication No. WO 92 / 09690; Ladner et al.International Publication No. WO 90 / 02809; Fuchs et al. (1991) Bio / Technology 9: 1370- 1372; Hay et al. (1992) Hum Antibody Hybridomas 3:81-85; Huse et al. (1989) Science 246: 1275-1281; Griffths et al. (1993) EMBO J 12:725-734; Hawkins et al. (1992) J Mol Biol 226:889-896; Clackson et al. (1991) Nature 352:624-628; Gram et al. (1992) PNAS 89:3576-3580; Garrad et al. (1991) Bio / Technology 9: 1373-1377; Hoogenboom et al.(1991) Nuc Acid Res 19:4133-4137; and Barbas et al. (1991) PNAS 88:7978-7982, each of which his hereby incorporated by reference in its entirety).

[0273] In some embodiments, an ANGPTL4 binding antibody or antigen-binding fragment thereof described herein may be derived from other species. A humanized antibody is an antibody produced by recombinant DNA technology, in which some or all amino acids of a human immunoglobulin light chain or heavy chain that are not required for antigen binding (e.g., constant regions and / or framework regions of variable domains) are used to substitute for the corresponding amino acids from light chain or heavy chain of the cognate, nonhuman antibody. By way of example, a humanized version of a murine antibody to a given antigen has on both heavy and light chains: (1) constant regions of a human antibody; (2) FRs from the variable domains of a human antibody; and (3) CDRs from the murine antibody. Human FRs may be selected based on their highest sequence homology to mouse FR sequence. When necessary, one or more residues in human FRs can be changed to residues at corresponding positions in a murine antibody so as to preserve binding affinity of the humanized antibody to a target. This change is sometimes called “back mutation.” Similarly, forward mutations may be made to revert back to murine sequence for a desired reason, e.g., stability or affinity to a target. Humanized antibodies generally are less likely to elicit an immune response in humans as compared to chimeric human antibodies because the former contain considerably fewer non-human components.

[0274] Methods for humanizing non-human antibodies are well known in the art. Suitable methods for making humanized antibodies in accordance with the present disclosure are described in, e.g., Winter EP 0 239 400; Jones et al., Nature 321 :522-525 (1986); Riechmann et al., Nature 332:323-327 (1988); Verhoeyen et al., Science 239: 1534- 1536 (1988); Queen et al., Proc. Nat. Acad. ScL USA 86: 10029 (1989); U.S. Patent 6,180,370; and Orlandi et al., Proc. Natl. Acad. Sd. USA 86:3833 (1989); the disclosures of each of which are incorporated herein by reference in their entireties. Generally, transplantation of non-human (e.g., murine) CDRs onto a human antibody is achieved as follows. cDNAs encoding VH and VL are isolated from a hybridoma, and nucleic acid sequences encoding VH and VL including CDRs are determined by sequencing. Nucleic acid sequences encoding CDRs are inserted into corresponding regions of a human antibody VH or VL coding sequences and attached to human constant region gene segments of a desired isotype (e.g., yl for CH and K for CL). Humanized heavy and light chain genes areco-expressed in mammalian host cells (e.g., CHO or NSO cells) to produce soluble humanized antibody. To facilitate large-scale production of antibodies, it is often desirable to select for a high expressor using, for example, a DHFR gene or GS gene in the producer line.

[0275] In some embodiments, an ANGPTL4 binding antibody or antigen-binding fragment thereof described herein comprises or is a human antibody. Completely human antibodies may be particularly desirable for therapeutic treatment of human subjects.Human antibodies can be made by a variety of methods known in the art including phage display methods described above using antibody libraries derived from human immunoglobulin sequences (see, e.g., U.S. Pat. Nos. 4,444,887 and 4,716,111; and PCT publications WO 98 / 46645, WO 98 / 60433, WO 98 / 24893, WO 98 / 16664, WO 96 / 34096, WO 96 / 33735, and WO 91 / 10741; each of which is incorporated herein by reference in its entirety) as well as Example 1 described herein. Techniques are also available for the preparation of human monoclonal antibodies in, e.g., Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Riss, (1985); and Boerner et al., J. Immunol., 147(1): 86-95, (1991), each of which is incorporated herein by reference in its entirety.Nucleic acids

[0276] The present disclosure, among other things, provides nucleic acids encoding ANGPTL4 binding antibodies or antigen-binding fragments thereof described herein. In some embodiments, nucleic acids provided herein delivers or provides an anti-ANGPTL4 antibody or antigen-binding fragment thereof. The present disclosure includes nucleic acids encoding one or more heavy chains, VH domains, heavy chain FRs, heavy chain CDRs, heavy chain constant domains, light chains, VL domains, light chain FRs, light chain CDRs, light chain constant domains, or other immunoglobulin-like sequences, antibodies, or antigen-binding fragments thereof disclosed herein. In some embodiments, nucleic acids encode polypeptides comprising segments or domains of the ANGPTL4-binding antibody chains described above. Some of the nucleic acids of the disclosure comprise the nucleotide sequence encoding the heavy chain variable region shown in SEQ ID NO: 13, 38, 58, 78, 98, 118, or 138, and / or the nucleotide sequence encoding the light chain variable region shown in SEQ ID NO: 23, 48, 68, 88, 108, 128, or 148. In a specific embodiment, the nucleic acidmolecules are those identified in Table 1. Some other nucleic acid molecules of the disclosure comprise nucleotide sequences that are substantially identical (e.g., at least 80%, 90%, 95%, or 99%) to the nucleotide sequences of those identified in Table 1. Some nucleic acid sequences of the disclosure comprise nucleotides encoding a heavy chain sequence that is substantially identical (e.g., at least 80%, 90%, 95%, or 99%) to the heavy chain sequence set forth in SEQ ID NO: 15, 28, 40, 60, 80, 100, 120, 140, 156, or 157. Some nucleic acid sequences comprising nucleotide encoding a light chain sequence that is substantially identical (e.g., at least 80%, 90%, or 99%) to the light chain sequence set forth in SEQ ID NO: 25, 50, 70, 90, 110, 130, or 150. Such nucleic acids may be present in a vector. Such nucleic acids may be present in the genome of a cell, e.g., a cell of a subject in need of treatment or a cell for production of an antibody, e.g., a mammalian cell for production of ANGPTL4 binding antibodies or antigen-binding fragments thereof described herein. When expressed from appropriate expression vectors, polypeptides encoded by these polynucleotides are capable of exhibiting ANGPTL4 antigen binding capacity.

[0277] Nucleic acids encoding ANGPTL4 binding antibodies or antigen-binding fragments thereof may be modified to include codons that are optimized for expression in a particular cell type or organism. Codon optimized sequences are synthetic sequences, and preferably encode an identical polypeptide (or biologically active fragment of a full length polypeptide which has substantially the same activity as the full length polypeptide) encoded by a non-codon optimized parent polynucleotide. In some embodiments, a coding region of a nucleic acids encoding ANGPTL4 binding antibodies or antigen-binding fragments thereof described herein, in whole or in part, may include an altered sequence to optimize codon usage for a particular cell type (e.g., a eukaryotic or prokaryotic cell). For example, a coding sequence for a humanized heavy (or light) chain variable region as described herein may be optimized for expression in a bacterial cell. Alternatively, the coding sequence may be optimized for expression in a mammalian cell (e.g., a CHO cell). Such a sequence may be described as a codon-optimized sequence.

[0278] Nucleic acid constructs of the present disclosure may be inserted into an expression vector or viral vector by methods known to the art, and nucleic acids may be operably linked to an expression control sequence. A vector comprising any nucleic acids or fragments thereof described herein is further provided by the present disclosure. Any nucleic acids or fragments thereof described herein can be cloned into any suitable vectorand can be used to transform or transfect any suitable host. Selection of vectors and methods to construct them are commonly known to persons of ordinary skill in the art (see, e.g., “Recombinant DNA Part D,” Methods in Enzymology, Vol. 153, Wu and Grossman, eds., Academic Press (1987)).

[0279] Conventionally used techniques including, for example, electrophoresis, calcium phosphate precipitation, DEAE-dextran transfection, or lipofection, may be used to introduce a foreign nucleic acid (e.g., DNA or RNA) into a prokaryotic or eukaryotic host cell. Desirably, a vector may include regulatory sequences, such as transcription and / or translation initiation and / or termination codons, which are specific to the type of host (e.g., bacterium, fungus, plant, or animal) into which a vector is to be introduced, as appropriate and taking into consideration whether a vector is DNA or RNA. In some embodiments, a vector comprises regulatory sequences that are specific to a genus of a host cell. In some embodiments, a vector comprises regulatory sequences that are specific to a species of a host.

[0280] In addition to a replication system and an inserted nucleic acid, a nucleic acid construct can include one or more marker genes, which allow for selection of transformed or transfected hosts. Exemplary marker genes include, e.g., biocide resistance (e.g., resistance to antibiotics or heavy metals) or complementation in an auxotrophic host to provide prototrophy.

[0281] An expression vector can comprise a native or nonnative promoter operably linked to an isolated or purified nucleic acid as described above. Selection of promoters, e.g., strong, weak, inducible, tissue-specific, and / or developmental-specific, is within the skill of one in the art. Similarly, combining a nucleic acid as described above with a promoter is also within the skill of one in the art.

[0282] Suitable vectors include those designed for propagation and expansion and / or for expression. For example, a cloning vector may be selected from the pUC series, the pBluescript series (Stratagene, LaJolla, Calif.), the pET series (Novagen, Madison, Wis.), the pGEX series (Pharmacia Biotech, Uppsala, Sweden), or the pEX series (Clontech, Palo Alto, Calif.). Bacteriophage vectors, such as ZGT10, ZGT11, ZZapII (Stratagene), ZEMBL4, and XNM1149, may be used. Examples of plant expression vectors that can be used include pBIHO, pBI101.2, pBI101.3, pBI121, or pBIN19 (Clontech). Examples of animal expression vectors that can be used include pEUK-Cl, pMAM, or pMAMneo (Clontech).The TOPO cloning system (Invitrogen, Carlsbad, Calif.) also can be used in accordance with the manufacturer's recommendations.

[0283] Additional sequences can be added to such cloning and / or expression sequences to optimize their function in cloning and / or expression, to aid in isolation of a nucleic acid encoding an ANGPTL4 binding antibody or antigen-binding fragment thereof described herein, or to improve introduction of a nucleic acid into a cell. Use of cloning vectors, expression vectors, adapters, and linkers is well known in the art (see, e.g., Sambrook et al., Molecular Cloning, a Laboratory Manual, 2d edition, Cold Spring Harbor Press, Cold Spring Harbor, N.Y. (1989); and Ausubel et al., Current Protocols in Molecular Biology, Greene Publishing Associates and John Wiley & Sons, New York, N.Y. (1994), each of which is hereby incorporated by reference in its entirety).

[0284] In some embodiments, nucleic acids and vectors of the present disclosure are isolated and / or purified. The present disclosure also provides a composition comprising an isolated or purified nucleic acid, optionally in the form of a vector. Isolated nucleic acids and vectors may be prepared using standard techniques known in the art including, for example, alkali / SDS treatment, CsCl binding, column chromatography, agarose gel electrophoresis, and / or other techniques well known in the art. The composition can comprise other components as described further herein.

[0285] Any method known to one skilled in the art for the insertion of nucleic acids into a vector may be used to construct expression vectors encoding an ANGPTL4 binding antibody or antigen-binding fragment thereof described herein under control of transcriptional and / or translational control signals. These methods may include in vitro recombinant DNA and synthetic techniques and in vivo recombination (see, e.g., Ausubel, supra; or Sambrook, supra).Methods of treatment

[0286] The present disclosure, among other things, provides methods of treating a disorder in a human comprising administering an antibody or antigen-binding fragment thereof described herein, thereby improving at least one sign or symptom of the disorder in the human after administration. In some embodiments, a therapeutically effective amount of at least one pharmaceutical composition described herein is administered to a human having a disorder.

[0287] In some embodiments, a disorder to be treated according to the present disclosure is or comprises a cardiovascular disease (CVD). In some embodiments, a CVD is or comprises atherosclerotic cardiovascular disease (ASCVD), a myocardial infarction (MI), stroke, coronary revascularization, arteriosclerosis, or any combination thereof. In some embodiments, a disorder is ASCVD.

[0288] CVD is a serious disease based on reduced survival, increased hospitalization, and impaired day-to-day functioning due to ischemia or infarct in the affected tissue. MI, stroke, and coronary revascularization in adults with established CVD represent a condition that is serious based on its impact on survival, day-to-day functioning, and the likelihood that the condition, if left untreated, will progress from a less severe condition to a more serious one.

[0289] Arteriosclerosis is a critical risk factor for CVD, and lipid metabolism disorders may be key contributors to its development. Arteriosclerosis is a degenerative process of the extracellular matrix of the arterial medial membrane, characterized by the thickening and hardening of the arterial wall due to structural and functional changes, resulting in a loss of elasticity and narrowing of the lumen. Atherosclerosis is primarily characterized by the formation of lipid-rich plaques in the arterial intima but can also sometimes present as aneurysms or ectasia.

[0290] Remnant cholesterol is highly atherogenic and may independently drive cardiovascular risk. Without wishing to be bound by any particular theory, it is proposed that elevated remnant cholesterol with metabolic risk factors drives high cardiovascular risk despite low LDL. Remnant cholesterol includes cholesterol carried by TRLs such as very low-density lipoproteins (VLDL), intermediate-density lipoproteins (IDL), and chylomicron remnants. From standard laboratory test, remnant cholesterol can be calculated total cholesterol minus HDL-C minus LDL-C (Figure 23). Remnant cholesterol particles may directly invade a vessel wall, similar to LDL. Unlike LDL, remnant cholesterol does not need to be oxidized to invade a vessel wall, which may contribute to their higher per particle atherogenicity than LDL. Remnant cholesterol particles may directly initiate inflammatory signaling in the endothelium as well, which is a contributor to atherogenesis.

[0291] In some embodiments, remnant cholesterol increases the production of reactive oxygen species. In some embodiment, reactive oxygen species lead to endothelial dysfunction.

[0292] In some embodiments, remnant cholesterol increases secretion of tumor necrosis factor-alpha (TNF-a) and interleukin (IL)-ip. In some embodiment, TNF-a and IL- ip increase apoptosis of endothelial cells promoting formation of atherosclerosis.

[0293] In some embodiments, remnant cholesterol promotes cytokine and pro- atherosclerotic factor production. In some embodiments, cytokine and pro-atherosclerotic factors promote white blood cell migration and promote inflammation leading to, e.g., formation of atherosclerosis.

[0294] In some embodiments, remnant cholesterol enhances platelet activity and / or aggregation by assembling a thromboplastin complex, upregulating a plasminogen activator inhibitor-1 gene, and / or expressing a plasminogen activator inhibitor-1 antigen.

[0295] In some embodiments, treatment with an antibody or antigen-binding fragment thereof that binds specifically to human ANGPTL4 according to the present disclosure reduces plasma remnant cholesterol levels. In some embodiments, treatment with an antibody or antigen-binding fragment thereof that binds specifically to human ANGPTL4 according to the present disclosure reduces increased remnant cholesterol. In some embodiments, treatment with an antibody or antigen-binding fragment thereof that binds specifically to human ANGPTL4 according to the present disclosure inhibits remnant cholesterol formation.

[0296] In some embodiments, treatment with an antibody or antigen-binding fragment thereof that binds specifically to human ANGPTL4 according to the present disclosure reduces plasma low-density lipoprotein cholesterol (LDL-C) levels. In some embodiments, treatment with an antibody or antigen-binding fragment thereof that binds specifically to human ANGPTL4 according to the present disclosure inhibits low-density lipoprotein cholesterol (LDL-C) formation.

[0297] In some embodiments, treatment with an antibody or antigen-binding fragment thereof that binds specifically to human ANGPTL4 according to the present disclosure reduces plasma triglyceride levels. In some embodiments, treatment with an antibody or antigen-binding fragment thereof that binds specifically to human ANGPTL4 according to the present disclosure inhibits triglyceride formation. In some embodiments, a human is afflicted with an elevated fasting plasma triglyceride concentration of at least about 150 mg / dl, such as at least about 160 mg / dl, at least about 170 mg / dl, at least about 190 mg / dl, at least about 190 mg / dl, at least about 200 mg / dL, at least about 300 mg / dL, atleast about 400 mg / dL, or at least about 500 mg / dL. In some embodiments, a human is afflicted with an elevated fasting plasma triglyceride concentration of at least about 150 mg / dl and less than about 880 mg / dl. In some embodiments, a human exhibits one, two, or three of: (1) remnant cholesterol levels greater than about 30 mg / dl, (2) low-density lipoprotein cholesterol (LDL-C) levels greater than or equal to about 50 mg / dL and less than or equal to about 130 mg / dL, and (3) stability on GLP1RA, lipid-lowering therapies. In some embodiments, a human is afflicted with an elevated fasting plasma triglyceride concentration of at least about 150 mg / dl, and optionally less than about 880 mg / dl, and optionally exhibits one, two, or three of: (1) remnant cholesterol levels greater than about 30 mg / dl, (2) low-density lipoprotein cholesterol (LDL-C) levels greater than or equal to about 50 mg / dL and less than or equal to about 130 mg / dL, and (3) stability on GLP1RA, lipid- lowering therapies.

[0298] In some embodiments, treatment with an antibody or antigen-binding fragment thereof that binds specifically to human ANGPTL4 according to the present disclosure reduces ectopic fat. In some embodiments, treatment with an antibody or antigenbinding fragment thereof that binds specifically to human ANGPTL4 according to the present disclosure prevent ectopic fat formation.

[0299] In some embodiments, treatment with an antibody or antigen-binding fragment thereof that binds specifically to human ANGPTL4 according to the present disclosure improve insulin sensitivity.

[0300] In some embodiments, a therapeutically effective amount of an ANGPTL4 binding antibody or antigen-binding fragment thereof described herein may be formulated and / or administered in a single dose. In some embodiments, a therapeutically effective amount of an ANGPTL4 binding antibody or antigen-binding fragment thereof may be formulated and / or administered in a plurality of doses, for example, as part of a dosing regimen.

[0301] In some embodiments, methods are provided for treating a human having a disorder, comprising subcutaneously administering a dose of about 15 mg to about 500 mg of an antibody or antigen-binding fragment thereof that binds specifically to human ANGPTL4 about weekly to about annually (e.g., about weekly, about biweekly, or about monthly) to the human, thereby improving at least one sign or symptom of the disorder in the human after administration.

[0302] A human to be treated with methods described herein can be e.g., a patient having, or at risk of having, a disease, disorder or condition described herein. A method of treating (e.g., one or more of reducing, inhibiting, or delaying progression of) a disorder with a pharmaceutical composition comprising at least one ANGPTL4 binding antibody or antigen-binding fragment thereof described herein is provided.

[0303] In some embodiments, an ANGPTL4 binding antibody or antigen-binding fragment is used to lower plasma triglyceride levels, such as lowering elevated plasma triglyceride levels in a subject with a disorder described herein. In some embodiments, an ANGPTL4 binding antibody or antigen-binding fragment is used to increase HDL levels and / or decrease non-HDL levels of cholesterol in a subject with a a disorder described herein.

[0304] An ANGPTL4 binding antibody or antigen-binding fragment of the present disclosure can be used, inter alia, to treat, prevent, and / or improve various disorders (e.g., ANGPTL4-associated disorders), including but not limited to any number of diseases, disorders, or conditions in which ANGPTL4 levels are aberrantly high and / or in which a reduction of ANGPTL4 levels is sought. These conditions include but are not limited to those involving lipid metabolism, such as hypercholesterolemia, hypertriglyceridemia, hyperlipidemia, hyperlipoproteinemia, dyslipidemia and lipodystrophy. In some embodiments, lipodystrophy includes, but is not limited to, familial lipodystrophy or acquired lipodystrophy. Familial partial lipodystrophy syndromes (FPL) are rare and highly morbid diseases. Estimated prevalence is between 1 : 100,000 and 1 : 10,000 worldwide for monogenic partial lipodystrophy. These disorders are characterized by selective loss of adipose tissue with insulin resistance and its concomitant metabolic complications including diabetes, dyslipidemia and hepatic steatosis. Patients are at extremely elevated risk for atherosclerotic cardiovascular disease, recurrent acute pancreatitis, cirrhosis, as well as significant impairments in quality of life and patient satisfaction.

[0305] In some embodiments, dyslipidemia includes, but is not limited to, atherogenic dyslipidemia, diabetic dyslipidemia, mixed dyslipidemia, hypertriglyceridemia, hypercholesterolemia, chylomicronemia, and / or other conditions caused by, e.g., decreased LPL activity and / or LPL deficiency, decreased LDL receptor activity and / or LDL receptor deficiency, altered ApoC2, ApoE deficiency, increased ApoB, increased production and / or decreased elimination of very low-density lipoprotein (VLDL), certain drug treatment (e.g.,glucocorticoid treatment-induced dyslipidemia), any genetic predisposition, diet, lifestyle, and the like. In some embodiments, hypertriglyceridemia includes, but is not limited to, severe hypertriglyceridemia (e.g., with plasma triglyceride concentration >500 mg / dL), hypertriglyceridemia associated with obesity, and type V hypertriglyceridemia.

[0306] In some embodiments, a disorder disclosed herein (e.g., an ANGPTL4- associated disorder) is associated with or resulting from one or more additional diseases, disorders, or conditions. Other disorders (e.g., ANGPTL4-associated disorders) associated with or resulting from, e.g., hyperlipidemia, hyperlipoproteinemia, and / or dyslipidemia, include, but are not limited to, obesity, fatty liver disease (e.g., nonalcoholic fatty liver disease), insulin resistance, metabolic syndrome, polycystic ovarian syndrome, chronic kidney disease, Cushing syndrome, hypercorti soli sm, acromegaly, heart failure, cardiovascular diseases or disorders,, acute pancreatitis, nonalcoholic steatohepatitis (NASH); and blood sugar disorders. In some embodiments, cardiovascular diseases or disorders include, but are not limited to, atherosclerosis, aneurysm, hypertension, angina, stroke, cerebrovascular diseases, congestive heart failure (e.g., heart failure with preserved ejection fraction), coronary artery diseases, myocardial infarction, peripheral vascular diseases, and the like. In some embodiments, bloods sugar disorders include, but are not limited to, diabetes (e.g., type 2 diabetes).

[0307] Lipodystrophy may be characterized by a complete or partial loss of and / or abnormal distribution of adipose tissue in certain areas of the body. It can be used as a general term for a group of conditions that are characterized by a complete (generalized) or partial loss of adipose tissue in certain areas of the body and / or abnormal distribution of fat tissue. There are multiple types of lipodystrophy, which may be caused by a genetic mutation or acquired (e.g., caused by another condition). The types of lipodystrophy affect people differently and have several different symptoms.

[0308] Familial partial lipodystrophy syndromes (FPL) are rare and highly morbid diseases. Estimated prevalence is between 1 : 100,000 and 1 : 10,000 worldwide for monogenic partial lipodystrophy. These disorders are characterized by selective loss of adipose tissue with insulin resistance and its concomitant metabolic complications including diabetes, dyslipidemia and hepatic steatosis. Patients are at extremely elevated risk for atherosclerotic cardiovascular disease, recurrent acute pancreatitis, cirrhosis, as well as significant impairments in quality of life and patient satisfaction.

[0309] In some embodiments, an ANGPTL4 binding antibody or antigen-binding fragment is used to lower fasting plasma insulin levels, such as lower elevated plasma insulin levels. In some embodiments, an ANGPTL4 binding antibody or antigen-binding fragment is used to reduce liver fat content. In some embodiments, an ANGPTL4 binding antibody or antigen-binding fragment is used to reduce gluteofemoral fat content.

[0310] In some embodiments, an ANGPTL4 antibody or antigen-binding fragment is used to reduce the risk of cardiometabolic diseases such as ASCVD, diabetes, NASH, and atherosclerotic cardiovascular disease. In some embodiments, an ANGPTL4 antibody or antigen-binding fragment is used to treat or reduce the risk of ASCVD. ASCVD may be caused by plaque buildup in arterial walls and may include, but is not limited to, diseases that comprise coronary heart disease (CHD), such as myocardial infarction, angina, and coronary artery stenosis; cerebrovascular disease, such as a transient ischemic attack, ischemic stroke, and carotid artery stenosis; peripheral artery disease, such as claudication; or aortic atherosclerotic disease, such as abdominal aortic aneurysm and ascending thoracic aneurysm. Currently, ASCVD-related conditions remain a leading cause of morbidity and mortality globally.

[0311] Administration of an ANGPTL4 binding antibody or antigen-binding fragment thereof described herein may be carried out in any convenient manner (e.g., injection, ingestion, transfusion, inhalation, implantation, or transplantation). In some embodiments, ANGPTL4 binding antibodies or antigen-binding fragments described herein are administered by injection or infusion. ANGPTL4 binding antibody or antigen-binding fragment described herein may be administered to a patient transarterially, subcutaneously, intravenously, intradermally, intratumorally, intranodally, intramedullary, intramuscularly, or intraperitoneally. In some embodiments, an ANGPTL4 binding antibody or antigenbinding fragment described herein is administered parenterally (e.g., intravenously, subcutaneously, intraperitoneally, or intramuscularly). In some embodiments, an ANGPTL4 binding antibody or antigen-binding fragment described herein is administered by subcutaneous, intravenous, intramuscular, or intrasternal infusion or injection. In some embodiments, an ANGPTL4 binding antibody or antigen-binding fragment described herein is administered by intramuscular or subcutaneous injection. ANGPTL4 binding antibody or antigen-binding fragment described herein may be injected directly into a site ofinflammation, a local disease site, a lymph node, an organ, a tumor, or site of infection in a human.

[0312] In some embodiments, at least one ANGPTL4 binding antibody or antigenbinding fragment described herein is utilized in combination with one or more other therapies, such as one or more additional therapeutic agents. Any suitable additional therapeutic agent may be administered with an antibody provided herein. The antibodies or antigen-bindings fragments thereof of the disclosure can also be used in combination with other agents for the prevention, treatment, or improvement of disorders. For example, statin therapies may be used in combination with the ANGPTL4 antibodies and / or antigen binding fragments of the invention for the treatment of patients with triglyceride-related disorders. In some embodiments, ANGPTL4 binding antibodies or antigen-binding fragments thereof described herein are administered in combination with plasma glucose lowering agents, such as, but not limited to metformin and GLP-1. In certain embodiments, patients undergoing standard diabetes and obesity treatment are also treated with ANGPTL4 binding antibody or antigen-binding fragment thereof described herein.

[0313] In some embodiments, combination therapy does not necessarily require that individual agents be administered together in a single composition (or even necessarily at the same time). In some embodiments, two or more therapeutic agents or modalities of a combination therapy are administered to a subject separately, e.g., in separate compositions, via separate administration routes (e.g., one agent orally and another agent intravenously), and / or at different time points. In some embodiments, two or more therapeutic agents may be administered together in a combination composition, or even in a combination compound (e.g., as part of a single chemical complex or covalent entity), via the same administration route, and / or at the same time.

[0314] In some embodiments, the second therapy is a diet (e.g., a low carbohydrate diet), exercise, and / or lifestyle changes.

[0315] In some embodiments, a second therapy is an LDL-C-lowering therapy. In some embodiments, a second treatment comprises treatment with proprotein convertase subtilisin / kexin type 9 inhibitors (PCSK9), ATP citrate lyase inhibitors, glucagon-like peptide-1 receptor agonists (GLP Ira), sodium-glucose cotransporter 2 (SGLT2) inhibitors, bempedoic acid, or any combination thereof. In some embodiment, a second therapycomprises incretin treatment. In some embodiment, a second therapy comprises statin treatment.Dosing

[0316] The methods described herein include administration of at least one dose of an antibody or antigen-binding fragment thereof that binds specifically to human ANGPTL4, e.g., a therapeutically effective dose of an ANGPTL4 binding antibody or antigen-binding fragment thereof and, optionally, one or more additional agents. An ANGPTL4 binding antibody or antigen-binding fragment thereof can be administered to a human in an amount sufficient to block LPL binding to ANGPTL4. An amount adequate to accomplish this is defined as a “therapeutically effective dose,” which may provide for an improvement or treatment of disorder.

[0317] Effective doses of ANGPTL4 binding antibodies or antigen-binding fragments thereof of the present disclosure for the treatment of diseases, disorders or conditions vary depending upon many different factors, including means of administration, target site, physiological state of the patient, whether the patient is human or an animal, other medications administered, and whether treatment is prophylactic or therapeutic.Usually, a patient is a human, but nonhuman mammals may also be treated, e.g., companion animals such as dogs, cats, horses, etc., laboratory mammals such as rabbits, mice, rats, monkeys, etc., and the like. Treatment dosages can be titrated to optimize safety and efficacy.

[0318] Administration of a therapeutically effective dose of an ANGPTL4 binding antibody or antigen-binding fragment thereof can be achieved in a number of different ways. Suitable administration of a therapeutically effective dose can entail administering a therapeutically effective dose in a particular regimen (e.g., a weekly regimen, a biweekly regimen, a 3 week regimen, a 4 week regimen, or a 5 week regime), such that a therapeutically effective dose is administered as a single dose or two or more doses (e.g., three, four, five, six, seven, eight or more doses) that are administered over a certain period of a particular regimen. Accordingly, a therapeutically effective dose can be split into multiple doses that combined do not exceed a particular regimen dose. Single or multiple administrations of the compositions may be administered depending on the dosage andfrequency as needed and tolerated by the patient. The particular dose used for a treatment will depend upon the medical condition and history of the mammal, as well as other factors such as age, weight, gender, administration route, efficiency, etc.

[0319] In some embodiments, an ANGPTL4 binding antibody or antigen-binding fragment thereof is administered as a single dose. In some embodiments, an ANGPTL4 binding antibody or antigen-binding fragment thereof is administered in a plurality of doses. In some embodiments, doses described herein are administered at least 2 or more times. In some embodiments, doses described herein are administered at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times or more. In some embodiments, doses and dosing frequencies described herein are repeated indefinitely.

[0320] In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about weekly to about annually. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about weekly to about biannually. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about weekly to about quarterly. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about weekly to about monthly.

[0321] In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered daily, semi-weekly, weekly, biweekly, once a month, and / or annually. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about biweekly. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about monthly. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about quarterly. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 orLTW980-HLE as described herein) is administered about biannually. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about annually.

[0322] In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 2 weeks to about every 30 weeks. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 2 weeks to about every 24 weeks. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 2 weeks to about every 12 weeks. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 2 weeks to about every 8 weeks. In some embodiments, an antibody or antigenbinding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980- HLE as described herein) is administered about every 4 weeks to about every 8 weeks.

[0323] In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every week, about every 2 weeks, about every 3 weeks, about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, about every 12 weeks, about every 13 weeks, about every 14 weeks, about every 15 weeks, about every 16 weeks, about every 17 weeks, about every 18 weeks, about every 19 weeks, about every 20 weeks, about every 21 weeks, about every 22 weeks, about every 23 weeks, about every 24 weeks, about every 25 weeks, about every 26 weeks, about every 27 weeks, about every 28 weeks, about every 29 weeks, or about every 30 weeks.

[0324] In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every week. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 2 weeks. In some embodiments, an antibodyor antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 3 weeks. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 4 weeks. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 5 weeks. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 6 weeks. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 7 weeks. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 8 weeks. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 9 weeks. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 10 weeks.

[0325] In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 11 weeks. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 12 weeks. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 13 weeks. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 14 weeks. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 15 weeks. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 16 weeks. In some embodiments, an antibodyor antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 17 weeks. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 18 weeks. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 19 weeks. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 20 weeks. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 21 weeks. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 22 weeks. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 23 weeks. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 24 weeks. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 25 weeks. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 26 weeks. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 27 weeks. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 28 weeks. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 29 weeks. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 30 weeks.

[0326] In some embodiments, bi-weekly administration of an ANGPTL4 binding antibody or antigen-binding fragment thereof comprises administering a particular dose as a single dose or splitting a particular dose into two or more doses that are administered to a patient over a period of two weeks (e.g., over a 1-5 day period). Accordingly, a particular biweekly dose can be split into multiple doses that combined do not exceed the particular biweekly dose (e.g., half of a particular dose can be administered to a patient on day 0 and the remaining half can be administered to the patient on day 7, or e.g., half of a particular dose can be administered on day 0 and the remaining half can be administered to the patient at day 1). In some embodiments, a bi-weekly dose of an ANGPTL4 binding antibody or antigen-binding fragment thereof can be administered on day 0, day 14, day 28, day 42, day 56, etc. As an example hereof, e.g., a bi-weekly dose of 400 mg can be administered on day 0, day 14, day 28, day 42, day 56, etc. In some embodiments, a bi-weekly dose of an ANGPTL4 binding antibody or antigen-binding fragment thereof can be administered by administering half of a bi-weekly dose to a patient on day 0 and administering the remaining half dose to a patient on one of the subsequent days (e.g., day 1, day 2, day 3, day 4, day 5, day 6, or day 7). In some embodiments, a bi-weekly dose of an ANGPTL4 binding antibody or antigen-binding fragment thereof can be administered by administering half of a biweekly dose to a patient on day 0 and administering the remaining half of a bi-weekly dose to a patient on day 1 and administering half of a bi-weekly dose to a patient on day 14 and administering the remaining half dose to a patient on day 15. As an example hereof, e.g., a bi-weekly dose of 400 mg can be administered as a 200 mg dose on day 0 and as a 200 mg dose on day 1, and as a 200 mg dose on day 14 and as a 200 mg dose on day 15.

[0327] In some embodiments, a bi-weekly dose of an ANGPTL4 binding antibody or antigen-binding fragment thereof can be administered by administering a third of a biweekly dose to a patient on day 0 and administering a third of a bi-weekly dose to a patient on day 1 and a third of a bi-weekly dose to a patient on day 2.

[0328] In some embodiments, a bi-weekly regimen is replaced with a weekly regimen, e.g., a weekly dose is administered once weekly or split into one or more doses that are administer to the patient over a one week period (e.g., over a 1-5 day period).

[0329] In some embodiments, dosing regimens described herein comprise replacing a bi-weekly regimen with a longer dosing regimen (e.g., a 3 week regimen, a 4 week regimen, or a 5 week regimen), such that a particular regimen dose is administered once orsplit into one or more doses that are administered to the patient over a certain time period (e.g., a 1-5 day period).

[0330] In some embodiments, an ANGPTL4 binding antibody or antigen-binding fragment thereof is administered in intervals of at least 2 days, such as at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, at least 14 days, or longer.

[0331] In some embodiments, an ANGPTL4 binding antibody or antigen-binding fragment thereof is administered in a dose of about 1 mg to about 1000 mg, such as of about 5 mg to about 900 mg, such as of about 10 mg to about 800 mg, such as of about 11 mg to about 750 mg, such as of about 12 mg to about 700 mg, such as of about 13 mg to about 650 mg, such as of about 14 mg to about 600 mg, such as of about 15 mg to about 500 mg, such as of about 20 mg to about 450 mg, such as of about 25 mg to about 400 mg, such as of about 50 mg to about 400 mg, such as of about 75 mg to about 300 mg, such as of about 100 mg to about 300 mg, such as of about 15 mg to about 150 mg, such as of about 150 mg to about 450 mg, such as of about 150 mg to about 400 mg, such as of about 150 mg to about 300 mg.

[0332] In some embodiments, an ANGPTL4 binding antibody or antigen-binding fragment thereof is administered about weekly to about annually (e.g., weekly, biweekly, or monthly) in a dose of about 1 mg to about 1000 mg, such as of about 5 mg to about 900 mg, such as of about 10 mg to about 800 mg, such as of about 11 mg to about 750 mg, such as of about 12 mg to about 700 mg, such as of about 13 mg to about 650 mg, such as of about 14 mg to about 600 mg, such as of about 15 mg to about 500 mg, such as of about 20 mg to about 450 mg, such as of about 25 mg to about 400 mg, such as of about 50 mg to about 400 mg, such as of about 75 mg to about 300 mg, such as of about 100 mg to about 300 mg, such as of about 15 mg to about 150 mg, such as of about 150 mg to about 450 mg, such as of about 150 mg to about 400 mg, such as of about 15 mg to about 300 mg.

[0333] Any dose described herein can be administered at any frequency described herein. For example, a dose range of about 15 mg to about 500 mg of an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) can be administered about weekly to about annually. Non-limiting examples of doses and frequencies include: a dose of about 15 mg of an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g.,LTW980 or LTW980-HLE as described herein) can be administered about every week to about every 12 weeks (e.g., about every 2 weeks, about every 4 weeks, or about every 8 weeks), a dose of about 50 mg of an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) can be administered about every week to about every 12 weeks (e.g., about every 2 weeks, about every 4 weeks, or about every 8 weeks), a dose of about 100 mg of an antibody or antigenbinding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980- HLE as described herein) can be administered about every week to about every 12 weeks (e.g., about every 2 weeks, about every 4 weeks, or about every 8 weeks), a dose of about 150 mg of an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) can be administered about every week to about every 12 weeks (e.g., about every 2 weeks, about every 4 weeks, or about every 8 weeks), a dose of about 200 mg of an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) can be administered about every week to about every 12 weeks (e.g., about every 2 weeks, about every 4 weeks, or about every 8 weeks), a dose of about 225 mg of an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) can be administered about every week to about every 12 weeks (e.g., about every 2 weeks, about every 4 weeks, or about every 8 weeks), a dose of about 300 mg of an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) can be administered about every week to about every 12 weeks (e.g., about every 2 weeks, about every 4 weeks, or about every 8 weeks), a dose of about 400 mg of an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) can be administered about every week to about every 12 weeks (e.g., about every 2 weeks, about every 4 weeks, or about every 8 weeks), a dose of about 450 mg of an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) can be administered about every week to about every 12 weeks (e.g., about every 2 weeks, about every 4 weeks, or about every 8 weeks), or a dose of about 500 mg of an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) can be administeredabout every week to about every 12 weeks (e.g., about every 2 weeks, about every 4 weeks, or about every 8 weeks).

[0334] In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every week at a dose of about 225 mg. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 2 weeks at a dose of about 225 mg. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 3 weeks at a dose of about 225 mg. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 4 weeks at a dose of about 225 mg. In some embodiments, an antibody or antigenbinding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980- HLE as described herein) is administered about every 5 weeks at a dose of about 225 mg. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 6 weeks at a dose of about 225 mg. In some embodiments, an antibody or antigenbinding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980- HLE as described herein) is administered about every 7 weeks at a dose of about 225 mg. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 8 weeks at a dose of about 225 mg. In some embodiments, an antibody or antigenbinding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980- HLE as described herein) is administered about every 9 weeks at a dose of about 225 mg. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 10 weeks at a dose of about 225 mg.

[0335] In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every week at a dose of about 300 mg. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g.,LTW980 or LTW980-HLE as described herein) is administered about every 2 weeks at a dose of about 300 mg. In some embodiments, an antibody or antigen -binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 3 weeks at a dose of about 300 mg. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 4 weeks at a dose of about 300 mg. In some embodiments, an antibody or antigenbinding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980- HLE as described herein) is administered about every 5 weeks at a dose of about 300 mg. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 6 weeks at a dose of about 300 mg. In some embodiments, an antibody or antigenbinding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980- HLE as described herein) is administered about every 7 weeks at a dose of about 300 mg. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 8 weeks at a dose of about 300 mg. In some embodiments, an antibody or antigenbinding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980- HLE as described herein) is administered about every 9 weeks at a dose of about 300 mg. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 10 weeks at a dose of about 300 mg.

[0336] In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every week at a dose of about 450 mg. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 2 weeks at a dose of about 450 mg. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 3 weeks at a dose of about 450 mg. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered aboutevery 4 weeks at a dose of about 450 mg. In some embodiments, an antibody or antigenbinding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980- HLE as described herein) is administered about every 5 weeks at a dose of about 450 mg. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 6 weeks at a dose of about 450 mg. In some embodiments, an antibody or antigenbinding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980- HLE as described herein) is administered about every 7 weeks at a dose of about 450 mg. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 8 weeks at a dose of about 450 mg. In some embodiments, an antibody or antigenbinding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980- HLE as described herein) is administered about every 9 weeks at a dose of about 450 mg. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 10 weeks at a dose of about 450 mg.

[0337] In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every week at a dose of about 900 mg. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 2 weeks at a dose of about 900 mg. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 3 weeks at a dose of about 900 mg. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 4 weeks at a dose of about 900 mg. In some embodiments, an antibody or antigenbinding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980- HLE as described herein) is administered about every 5 weeks at a dose of about 900 mg. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 6 weeks at a dose of about 900 mg. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980- HLE as described herein) is administered about every 7 weeks at a dose of about 900 mg. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 8 weeks at a dose of about 900 mg. In some embodiments, an antibody or antigenbinding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980- HLE as described herein) is administered about every 9 weeks at a dose of about 900 mg. In some embodiments, an antibody or antigen-binding fragment thereof that binds specifically to ANGPTL4 (e.g., LTW980 or LTW980-HLE as described herein) is administered about every 10 weeks at a dose of about 900 mg.

[0338] A therapeutically effective dose of the ANGPTL4 binding antibody or antigen-binding fragment thereof can be, for example, about 0.5 mg / kg body weight or more (e.g., about 1 mg / kg or more, about 2 mg / kg or more, about 3 mg / kg or more, about 4 mg / kg or more, about 5 mg / kg or more, about 6 mg / kg or more, or about 7 mg / kg or more, or about 8 mg / kg or more, or about 9 mg / kg or more, or about 10 mg / kg or more), or from about 0.5 mg / kg to about 10 mg / kg (e.g., from about 1 mg / kg to about 9 mg / kg, or from about 2 mg / kg to about 8 mg / kg).

[0339] A dose of an ANGPTL4 binding antibody or antigen-binding fragment thereof can be a single dose. For example, a single dose can be given irrespective of a particular subject’s weight. A single dose can be, e.g., 15-1000, 75-450, 150-300, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 33, 340,350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 510, 520,530, 540, 550, 560, 570, 580, 590, 600, 620, 640, 660, 680, 700, 720, 740, 760, 780, 800,850, 900, 950, 1000 mg, or an interim number of mg thereof.Dosing cycles

[0340] A method of treating a human having a disorder can include administering a dose of about 15 mg to about 500 mg of an antibody or antigen-binding fragment thereof that binds specifically to human ANGPTL4. In some embodiments, the ANGPTL4 binding antibody or antigen-binding fragment is administered about weekly to about annually (e.g., weekly, biweekly, or monthly).

[0341] Administration can occur in one or more cycles, for example, a first cycle can have a first dosing scheme and one or more subsequent cycles can have dosing scheme(s) that are distinct from (or the same as) the first cycle. A first dosing cycle may include a number of doses at a particular concentration given e.g., once every week, once every 2 weeks, or once every 3 weeks. A second dosing cycle may include a number of doses at a particular concentration given e.g., once every week, once every 2 weeks, or once every 3 weeks. The concentration of ANGPTL4 binding antibody or antigen-binding fragment in a dose may be the same in each dosing cycle or may differ between the first dosing cycle and the second dosing cycle. The dosing schedule may be the same in each dosing cycle or it may differ between the first dosing cycle and the second dosing cycle.

[0342] An ANGPTL4 binding antibody or antigen-binding fragment thereof can be administered to a subject in a given cycle. In some embodiments, an ANGPTL4 binding antibody can be administered to a subject in a given cycle, e.g., once every week, once every 2 weeks, or once every 3 weeks as a dose ranging from about 1 mg to about 1000 mg ANGPTL4 antibody or antigen-binding fragment thereof, such as such as of about 5 mg to about 900 mg, such as of about 10 mg to about 800 mg, such as of about 11 mg to about 750 mg, such as of about 12 mg to about 700 mg, such as of about 13 mg to about 650 mg, such as of about 14 mg to about 600 mg, such as of about 15 mg to about 500 mg, such as of about 20 mg to about 450 mg, such as of about 25 mg to about 400 mg, such as of about 50 mg to about 400 mg, such as of about 75 mg to about 300 mg, such as of about 100 mg to about 300 mg, such as of about 15 mg to about 150 mg, such as of about 150 mg to about 450 mg, such as of about 150 mg to about 400 mg, such as of about 150 mg to about 300 mg.

[0343] An ANGPTL4 binding antibody or antigen-binding fragment thereof can be administered to a subject in a first cycle followed by second cycle, wherein the dose administered in the second cycle differs from the dose administered in the first cycle. The first cycle can be about 1 week to about 12 weeks in duration, such as about 2 weeks to about 10 weeks, such as about 3 weeks to about 8 weeks, such as about 2 weeks to about 6 weeks in duration. The first cycle can be about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, or about 10 weeks in duration. The second cycle can be about 1 week to about 12 weeks in duration, such as about 2 weeks to about 10 weeks, such as about 3 weeks to about 8 weeks,such as about 2 weeks to about 6 weeks in duration. The second cycle can be about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, or about 10 weeks in duration.

[0344] An ANGPTL4 binding antibody or antigen-binding fragment thereof can be administered to the subject in the first cycle once every second week at a dose of about 15 mg to about 500 mg of an antibody or antigen-binding fragment thereof that binds specifically to human ANGPTL4.

[0345] An ANGPTL4 binding antibody or antigen-binding fragment thereof can be administered in a second cycle comprising a dose of about 15 mg to about 500 mg of an antibody or antigen-binding fragment thereof that binds specifically to human ANGPTL4 once every 2 weeks. In some embodiments, the dose administered in the first cycle and the second cycle is the same. In some embodiments, the dose administered in the second cycle differs from the dose administered in the first cycle.

[0346] Additional cycles can be used. For example, at least one additional cycle, optionally 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or greater than 20 additional cycles can be used. The dosing regimen of the at least one additional cycle can be the same or different as the first or second cycle, optionally wherein the ANGPTL4 binding antibody or antigen-binding fragment thereof portion of the dosing regimen is discontinued after completing 6 total cycles.

[0347] Also disclosed herein is a method of treating a human having a disorder, comprising administering an ANGPTL4 binding antibody or antigen-binding fragment thereof to the human for at least two distinct cycles of four to eight weeks each, the first cycle comprising administering a first dose of ANGPTL4 binding antibody at 150 mg antibody, and the second cycle comprising administering a second dose of at least 450 mg of ANGPTL4 binding antibody or antigen-binding fragment thereof once every two weeks.

[0348] In some embodiments, the step of administering the ANGPTL4 binding antibody or antigen-binding fragment thereof results in a reduction of fasting plasma triglyceride levels of at least 10%, such as at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%, compared to basal fasting plasma triglyceride levels.

[0349] In some embodiments, the step of administering the ANGPTL4 binding antibody or antigen-binding fragment thereof results in a reduction of postprandial plasma triglyceride levels of at least 10%, such as at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%, compared to basal postprandial plasma triglyceride levels.Pharmaceutical composition

[0350] The present disclosure, among other things, provides pharmaceutical compositions comprising at least one ANGPTL4 binding antibody or antigen-binding fragment in combination with one or more pharmaceutically or physiologically acceptable carriers, diluents, or excipients.

[0351] When “a therapeutically effective amount” is indicated, a precise amount of a pharmaceutical composition comprising at least one ANGPTL4 binding antibody or antigenbinding fragment described herein can be determined by a physician with consideration of individual differences in age, weight, immune response, and condition of the patient (human).

[0352] Pharmaceutical compositions described herein may comprise buffers including neutral buffered saline or phosphate buffered saline (PBS); carbohydrates, such as glucose, mannose, sucrose, dextrans, or mannitol; proteins, polypeptides, or amino acids (e.g., glycine); antioxidants; chelating agents, such as EDTA or glutathione; adjuvants (e.g., aluminum hydroxide); and preservatives. In some embodiments, a pharmaceutical composition is substantially free of contaminants, e.g., there are no detectable levels of a contaminant (e.g., an endotoxin).

[0353] Pharmaceutical compositions described herein may be in a variety of forms. These include, for example, liquid, semi-solid and solid dosage forms, such as liquid solutions (e.g., injectable and infusible solutions), dispersions or suspensions, liposomes, and suppositories. Preferred compositions may be injectable or infusible solutions. Pharmaceutical compositions described herein can be formulated for administration intravenously, subcutaneously, intradermally, intratumorally, intranodally, intramedullary, intramuscularly, transarterially, or intraperitoneally. In some embodiments, the ANGPTL4 binding agent is administered subcutaneously.

[0354] In some embodiments, a pharmaceutical composition described herein is formulated for parenteral (e.g., intravenous, subcutaneous, intraperitoneal, or intramuscular) administration. In some embodiments, a pharmaceutical composition described herein is formulated for subcutaneous, intravenous, intramuscular, or intrasternal injection or infusion. In preferred embodiments, a pharmaceutical composition described herein is formulated for subcutaneous or intravenous injection of infusion. Pharmaceutical compositions described herein can be formulated for administered by using infusion techniques that are commonly known in immunotherapy (See, e.g., Rosenberg et al., New Eng. J. of Med. 319: 1676, 1988, which is hereby incorporated by reference in its entirety).

[0355] As used herein, the terms “parenteral administration” and “administered parenterally” refer to modes of administration other than enteral and topical administration, usually by injection or infusion, and include, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, intratumoral, and intrasternal injection and infusion.

[0356] Pharmaceutical compositions comprising at least one ANGPTL4 binding antibody or fragment thereof described herein can be administered to a human in accordance with a dosage regimen described herein, alone or in combination with one or more therapies. Pharmaceutical compositions comprising at least one ANGPTL4 binding antibody or antigen-binding fragment described herein can be for use in the manufacture of a medicament for treating a disorder in a human.

[0357] In some embodiments, pharmaceutical compositions described herein are administered in combination with plasma glucose lowering agents, such as, but not limited to metformin, and GLP-1. In certain embodiments, human undergo standard diabetes and obesity treatment are also treated with ANGPTL4 binding antibody or antigen-binding fragment described herein.Kit

[0358] The present disclosure, among other things, provides kits comprising at least one ANGPTL4 binding antibody or antigen-binding fragment described herein, and instructions for use and / or administration. In some embodiments, a kit comprises least oneANGPTL4 binding antibody or antigen-binding fragment described herein and a pharmaceutically acceptable carrier, and instructions for use and / or administration.

[0359] Also provided are kits for use in the various methods disclosed herein. In some embodiments, a kit comprises instructions for use in any method described herein. Instructions can comprise a description of administration of the first and second pharmaceutical compositions to a subject to achieve the intended activity in a subject. The kit may further comprise a description of selecting a human suitable for treatment based on identifying whether the human is in need of the treatment. In some embodiments, the instructions comprise a description of administering at least one ANGPTL4 binding antibody or antigen-binding fragment to a subject who is in need of the treatment.

[0360] The instructions relating to administering a dose comprising at least one ANGPTL4 binding antibody or antigen-binding fragment described herein generally include information as to dosage, dosing schedule, and route of administration for the intended treatment. The containers may be unit doses, bulk packages (e.g., multi -dose packages) or sub-unit doses. Instructions supplied in the kits of the disclosure are typically written instructions on a label or package insert. The label or package insert indicates that the pharmaceutical compositions are used for treating, delaying the onset, and / or alleviating a disease, disorder or condition in a subject.

[0361] The kits provided herein are in suitable packaging. Suitable packaging includes, but is not limited to, vials, bottles, jars, flexible packaging, and the like. Also contemplated are packages for use in combination with a specific device, such as an infusion device. A kit may have a sterile access port (for example, the container may be an intravenous solution bag or a vial having a stopper pierce able by a hypodermic injection needle). The container may also have a sterile access port.

[0362] Kits optionally may provide additional components such as buffers and interpretive information. Normally, the kit comprises a container and a label or package insert(s) on or associated with the container. In some embodiment, the disclosure provides articles of manufacture comprising contents of the kits described above.EXEMPLIFICATION

[0363] The following examples are provided so as to describe to the skilled artisan how to make and use methods and compositions described herein, and are not intended to limit the scope of the present disclosure.Example 1: Preparation of ANGPTL4 proteins and ANGPTL4 binding antibodies (LTW980)

[0364] The present example demonstrates that LTW980 (a monoclonal antibody as described in the Examples herein) specifically binds to human ANGPTL4. The present example also documents that LTW980 prevents LPL inhibition by human ANGPTL4 and cynomolgus monkey ANGPTL4, thus the example confirms that LTW980 is a potent LPL inhibitor antibody.Material and methodsPreparation of non-biotinylated and biotinylated recombinant ANGPTL4-FLAG-6HIS-Avi proteins

[0365] A nucleic acid sequence encoding full-length human ANGPTL4 polypeptide (amino acids 26-406, matching NCBI sequence NM_139314.2) with N-terminal signal peptide from human IgG-kappa (MKTFILLLWVLLLWVIFLLPGATA, SEQ ID NO: 152), and C-terminal FLAG epitope (DYKDDDDK, SEQ ID NO: 153), hexahistidine purification tag (HHHHHH, SEQ ID NO: 154), and Avi tag (i.e., BirA biotinylation sequence GGGLNDIFEAQKIEWHE, SEQ ID NO: 155) was subcloned into the mammalian cell expression vector pRS5a to generate the plasmid pRS-Ikk-hANGPTL4(26-406)-FLAG- 6HIS-Avi.

[0366] Suspension-adapted HEK293T cells were cultured in serum-free FreeStyle 293 expression medium and transfected with the plasmid pRS-Ikk-hANGPTL4 (26-406)- FLAG-6HIS-Avi using polyethyleneimine as the transfection reagent. Five hours after transfection, heparin was added to the culture medium to a final concentration of 0.5 mg / mL. The cells were cultured for 72-96 hours and the supernatant was harvested by centrifugation at 4 °C and sterile-filtered using a 0.22 pm filter. The filtered cell culture supernatant was then concentrated to about 100 mL by tangential flow filtration (TFF). The concentrated supernatant was diluted to a volume of 1 liter with TBS-glycerol buffer (50 mM Tris-HCl, 150 mM NaCl, 15% (v / v) glycerol, pH 7.4) and the sample was concentratedto about 200 mL by TFF. Anti -Flag M2 agarose resin pre-equilibrated with TBS-glycerol buffer was then added to the sample, and the resulting solution was gently mixed for 1 hour at 4 °C. The agarose resin was then washed 5 times with 25 mL TBS-glycerol, and the bound ANGPTL4 protein was eluted with 20 mL TBS-glycerol containing 0.2 mg / mL Flag peptide. Peroxide-free Tween-20 was added to the eluted protein solution to a final concentration of 0.1%, and the resulting solution was loaded onto a 5 mL HiTrap heparin column that was pre-equilibrated in TBS-glycerol containing 0.1% Tween-20 (Buffer A). The column was washed with 50 mL Buffer A, followed by 50 mL Buffer A containing 300 mM NaCl. ANGPTL4 protein was then eluted with 20 mL Buffer A containing 600 mM NaCl. The eluted protein was concentrated using a centrifugal concentrator with a 30 kDa molecular weight cutoff. The purity of the purified ANGPTL4 protein as assessed by SDS- PAGE was >90%. The same methods were used to prepare cynomolgus monkey, mouse and rat ANGPTL4 proteins.

[0367] For some applications, ANGPTL4 proteins were site-specifically biotinylated on the C-terminal Avi tag using 10 pg purified biotin-protein ligase (BirA, Avidity) per mg of ANGPTL4. The buffer was supplemented with final concentrations of 10 mM ATP, 10 mM magnesium acetate, and 0.5 mM d-biotin. The reaction mixture was incubated for 2 hours at 30 °C and overnight at 4 °C, then loaded onto a HiLoad Superdex 200 column (26 mm x 600 mm) that was equilibrated in Buffer A. Fractions from the Superdex 200 column were analyzed using SDS-PAGE, and ANGPTL4 containing fractions were pooled and concentrated using a centrifugal concentrator.Generation ofANGPTL4 antibody LTW980

[0368] Recombinant human ANGPTL4 protein was prepared as described above and used as immunogen for the generation of anti-ANGPTL4 hybridoma clones. Bcl-2 transgenic mice were immunized with recombinant human ANGPTL4 according to a standard rapid immunization protocol. Hybridomas were generated by using a standard electrofusion-based method. CHO-K1PD cells stably expressing human ANGPTL4 fused to a transmembrane domain were generated using standard methods. Due to the presence of the transmembrane domain, these cells display ANGPTL4 on the cell surface. Therefore, binding of antibodies to ANGPTL4 on the surface of these cells can be detected using flow cytometry.

[0369] Hybridoma supernatants were screened by detecting binding of antibodies present in the supernatant to human ANGPTL4 expressed on the surface of CHO-K1PD cells. Binding of antibodies to the cells was detected using a fluorescently labeled antimouse secondary antibody and flow cytometry. Parental CHO-K1PD cells that do not express ANGPTL4 were used as a negative control. For hybridomas that bound to ANGPTL4, antibodies were purified from cell supernatants using standard methods, and the resulting enriched supernatant was tested in the flow cytometry assay with CHO- K1PD / ANGPTL4 and CHO-KIPD-Parental cells.

[0370] ANGPTL4 antibody titers in hybridoma supernatants were determined by using a standard direct ELISA assay, in which recombinant human ANGPTL4 protein was immobilized on the surface of the ELISA plate. Confirmed positive hybridomas were subcloned, and the sequences of the monoclonal antibodies produced by these hybridomas was determined using standard methods.

[0371] Some of the resulting monoclonal antibodies, including the precursor to LTW980, 19C16, were subsequently shown to inhibit human ANGPTL4 mediated inhibition of human LPL using methods described below. The nucleotide and amino acid sequences of the heavy and light chain variable regions of 19C16 was determined using standard methods. 19C16 was humanized using standard methods and the resulting humanized antibody was affinity matured by using error-prone PCR and yeast display technology to yield LTW980. A CHO cell line stably expressing LTW980 was prepared and LTW980 was purified from the supernatant of these cells using Protein A affinity chromatography.Human LPL enzyme activity assay using human VLDL as the substrate

[0372] Recombinant human LPL was prepared using the following procedure. HEK293T cells cultured in FreeStyle 293 expression medium were transfected with a mammalian expression plasmid encoding full-length human LPL polypeptide (matching NCBI sequence NM_000237.2) using a standard polyethyleneimine transfection method. At 24 hours after transfection, heparin was added to the culture medium to a final concentration of 3 U / mL, to enhance release of secreted LPL from the cell surface. At 60 hours posttransfection, the culture medium was collected, filtered using a 0.2 pm filter, and glycerol was added to a final concentration of 10% (v / v). The resulting solution was loaded onto a 5mL Heparin Sepharose HiTrap column which had been pre-equilibrated with Buffer A (50 mM Tris-HCl, 200 mM NaCl, containing 10% (v / v) glycerol, pH 7.2). The column was washed with Buffer A, and human LPL protein was then eluted with step gradients of 500 mM NaCl, IM NaCl, and 2M NaCl in Buffer A. Elution fractions were assayed for LPL enzyme activity using the fluorogenic substrate l,2-o-dilauryl-rac-glycero-3 -glutaric acid- (6'-methylresorufin) ester (DGGR), and protein purity was assessed by SDS-PAGE. The most catalytically active and highest purity LPL eluted with 2M NaCl. Aliquots of purified human LPL were flash-frozen and stored at -80 °C until use.

[0373] The following protocol was used to assess the ability of LTW980 to prevent ANGPTL4 inhibition of human LPL. A 384-well assay plate and sample plate were each washed with 1% (w / v) BSA (0.1 mL per well) for 30 minutes at room temperature. The plates were then washed twice with DPBS containing 0.05% (v / v) Tween-20 solution. ANGPTL4 antibody in 100 mM HEPES, pH 7.0 (20 pL per well, serial dilution with final assay concentrations ranging from 0.02 to 500 nM) was added to the sample plate, followed by 20 pL human ANGPTL4 protein (10 nM final assay concentration; sufficient to inhibit LPL enzyme activity by 70-95%) in Assay Buffer (100 mM HEPES, 2 mM MgC12, pH 7.0), and the plate was incubated for 20 minutes at room temperature with gentle shaking. LPL diluted in Assay Buffer (20 pL) was then added and the plate was incubated for 10 minutes at room temperature with gentle shaking. A Coupling Enzyme Mix was prepared containing 2.7 U / mL acyl-coenzyme A oxidase, 6.7 U / mL acyl-coenzyme A synthetase, 6.4 U / mL horseradish peroxidase, 4 mM ATP and 0.5 mM coenzyme A, and catalase agarose bead suspension (one-fifth of final solution volume) in Assay Buffer. The mixture was incubated at 4 °C for 30 minutes with shaking, after which the catalase agarose beads were removed by centrifugation at 3,000 rpm for 5 minutes at 4 °C, and transferring the supernatant to a new tube. A human VLDL / Amplex Red solution with final concentrations of 12.3 pg / mL human VLDL and 33 pM Amplex Red was prepared by diluting human VLDL stock solution (1.4 mg / mL) in Assay Buffer and adding catalase beads suspension (one-fifth of final solution volume). The mixture was then incubated at 4 °C for 30 minutes, and the beads removed from the solution by centrifugation. Amplex Red diluted in Assay Buffer was then added. To the solution in the sample plate containing LPL, ANGPTL4 and ANGPTL4 antibody (60 pL), Coupling Enzyme Mix (20 pL) was added, and 54 pL of the resulting solution was transferred to the assay plate. To initiate the LPL reaction, VLDL / Amplex Red solution (18pL) was added, and resorufin fluorescence was monitored continuously for 30 minutes using an EnVision multiwell plate reader (Perkin Elmer). Final assay concentrations were: 10 nM ANGPTL4, ~4 nM human LPL, 2.3 pg / mL human VLDL, 0.75 mM ATP, 90 pM coenzyme A, 0.5 U / mL ACO, 1.25 U / mL ACS, 1.2 U / mL HRP, and 10 pM Amplex Red. The resulting resorufin fluorescence over time data was used to determine LPL enzyme activity (initial rate) for each sample. Control samples without LPL, or without ANGPTL4 and ANGPTL4 antibody, were used to normalize the enzyme activity, which was expressed as a percentage of LPL activity in the absence of ANGPTL4 and ANGPTL4 antibody. Enzyme activity data for different ANGPTL4 antibody concentrations were plotted using GraphPad Prism software, and using the data fitting function an EC50 value for the ANGPTL4 antibody-mediated increase in LPL enzyme activity was generated.LTW980 binding affinities assessed by solution equilibrium titration (SET)

[0374] SET assays were performed as follows. In a 96-well polypropylene plate, a constant concentration of ANGPTL4 antibody (10 pM) was mixed with different concentrations of non-biotinylated human, cyno, mouse, or rat full-length ANGPTL4 protein (5-fold serial dilution ranging from 0.01 pM to 100 nM) in SET buffer (PBS, pH 7.4 without CaC12 or MgC12, containing 0.5% (w / v) fatty acid-free BSA and 0.02% (v / v) Tween-20). The final reaction volume was 80 pL. The plate was sealed using an adhesive film and incubated at 22 °C for 14 hours with constant shaking (300 rpm). In parallel, a 384-well streptavidin-coated Meso Scale Discovery (MSD) plate was blocked by incubating the plate with 50 pL blocking buffer (PBS, pH 7.4, containing 5% (w / v) BSA) per well at 4 °C. The blocked MSD plate was washed 3 times with wash buffer (PBS, pH 7.4 containing 0.05% (v / v) Tween-20) using a plate washer (BioTek). Biotinylated ANGPTL4 protein (1 nM, 15 pL per well) was immobilized on the surface of the streptavidin-coated MSD plate by incubation at 22 °C for 1 hour with constant shaking (600 rpm). The plate was then washed 3 times as described previously.

[0375] The equilibrium binding reactions (15 pL per well) were applied to the MSD plate with immobilized ANGPTL4 and incubated for 20 minutes at 22 °C. The unbound material was removed by washing the plate 3 times with wash buffer, and the captured antibody was detected by adding 15 pL per well of a 1 :500 dilution of Sulfo-tagged goat anti-human IgG (Meso Scale Discovery). The plate was then incubated for 1 hour withconstant shaking (600 rpm). The plate was washed 3 times, and then 15 pL / well of lx MSD read buffer T was added and the plate was developed using a Sector Imager 6000 (Meso Scale Discovery). The data were transferred to Microsoft Excel for analysis and plotted using GraphPad Prism v6. The KD values were determined by fitting the data to the following equation: y = (Bmax / (CAb / 2))*((CAb / 2)-((((((CAg+CAb)+KD) / 2)-((((((CAg+CAb)+KD)A2) / 4)- (CAg*CAb))A0.5))A2) / (2*CAb))), where Bmax is the signal when no ANGPTL4 protein is present in solution, C \b is the constant concentration of ANGPTL4 antibody in solution, C \gis the concentration of ANGPTL4 in solution, and KD is the equilibrium dissociation constant.LTW980 binding affinities assessed by ForteBio Octet kinetic binding assay

[0376] Association rate constants (ka), dissociation rate constants (kd), and equilibrium dissociation constants (KD) were determined by using an Octet (ForteBio) kinetic binding assay. 10X Kinetics Buffer (ForteBio) was diluted 10-fold with DPBS, and the resulting IX Kinetics Buffer was added (0.2 mL per well) to a 96-well plate.Streptavidin sensors (ForteBio) were immersed in the solution and equilibrated for at least 10 minutes at room temperature. In a second 96-well plate, sensors were washed in IX Kinetics buffer, and then immersed in 200 pL of 25 nM biotinylated human ANGPTL4, biotinylated ANGPTL3, or biotinylated reference protein (for background subtraction) for 1000 seconds at room temperature. The sensors were then washed in IX Kinetics buffer for 120 seconds, and immersed in 200 pL of ANGPTL4 or ANGPTL3 antibody diluted in IX Kinetics buffer at various concentrations (serial 2-fold dilutions; the highest concentrations were 12.5 nM or 25 nM; the lowest concentrations ranged from 0.8 to 3 nM; 4-6 different antibody concentrations were used for each KD determination), and antibody association was monitored for 480 seconds. The sensors were then transferred to a well containing 200 pL IX Kinetics buffer, and antibody dissociation was monitored for 1200 seconds. Background-corrected association and dissociation curves were globally fitted by Octet Software (ForteBio) to generate ka and kd values, from which KD was calculated using the equation KD = kd / ka.Results

[0377] LTW980 bound to recombinant human ANGPTL4 and cynomolgus monkey ANGPTL4 with high affinity (KD = 10 pM and 7 pM, respectively, as assessed by solution equilibrium titration (SET) assay, and KD = 10 pM and 21 pM, respectively, as assessed by ForteBio assay) but did not bind with similarly high affinity to mouse ANGPTL4 or rat ANGPTL4 (KD > 800 pM as assessed by SET assay, and KD = 6200-6340 pM as assessed by ForteBio) (Table 2, Table 3).Table 2. LTW980 binds to human and monkey ANGPTL4 with high affinity. Affinities were determined by using a solution equilibrium titration (SET) assay with recombinant, full-length ANGPTL4 proteins.Table 3. LTW980 binds to human ANGPTL4 and monkey ANGPTL4 with high affinity and does not bind to human ANGPTL3. Binding affinities were determined by using a ForteBio Octet kinetic binding assay.

[0378] In a biochemical LPL inhibition assay using recombinant LPL with human very low density lipoprotein (VLDL) as the substrate, LTW980 prevented LPL inhibition by human and cynomolgus monkey ANGPTL4 with IC50 values of 1.5 nM and 2.2 nM, respectively (Figure 12, Table 4).Table 4. LTW980 prevents LPL inhibition by human ANGPTL4 and cynomolgus monkey ANGPTL4.Reported data are mean values ± standard deviation (SD) from four replicate experiments. ANGPTL4 proteins were used at a final concentration of 10 nM at which concentration it inhibited LPL enzyme activity by 70-95% in the absence of ANGPTL4.

[0379] In summary, LTW980 is a humanized ANGPTL4 antibody that binds to human and cynomolgus monkey ANGPTL4 with high affinity and prevented ANGPTL4inhibition of LPL in a cell-free LPL enzyme activity assay. Hereby showing that LTW980 is a potent LPL inhibitor antibody.Example 2: Efficacy and safety studies in monkeys

[0380] The present example demonstrates that LTW980 lowered plasma triglycerides and remnant cholesterol in hypertriglyceridemic monkeys.Material and methods

[0381] ANGPTL4 proteins and ANGPT antibodies were prepared as described in Example 1.LTW980 effect on plasma TG in hypertriglyceridemic monkeys

[0382] Cynomolgus monkeys used in the study were cared for in accordance with institutional guidelines. The study protocol was reviewed and approved by the Institutional Animal Care and Use Committee (IACUC) of the facility conducting the study.Hypertriglyceridemic cynomolgus monkeys were administered a single subcutaneous dose of LTW980 (3 mg / kg). Plasma total LTW980 concentrations and plasma triglyceride levels were then monitored at various timepoints; triglycerides were measured in the morning prior to the animals' morning feeding.

[0383] Four male, diabetic, hypertriglyceridemic, cynomolgus monkeys (Macaca fascicularis) were used in this pharmacology study; all animals received once daily injections of Lantus insulin throughout the study. The animals were individually housed in a humidity (60%; range 30-90%) and temperature-controlled (76 °F; range 72-80 °F) environment, maintained on a 12-h light / dark cycle, and provided socialization and enrichment activities daily. Animals were fed standard monkey chow diet (LabDiet 5047 PMI, Richmond, IN) supplemented with fresh fruits, vegetables, and peanuts / almonds. Animals were allowed access to purified water ad libitum. A 30.3 mg / mL stock solution of LTW980 (ID no. LTW980-13882A-AK057-UFT001-W2) in 10 mM L-Histidine / Histidine- HC1 (pH 5.0) buffer was shipped frozen to the test site, stored at -80°C, and thawed under refrigeration starting 3 days prior to use. On the morning of dosing, pre-calculated volumes were drawn up into dosing syringes (3 mL syringe and 25 G needle) and kept at room temperature until administration. Animals were restrained in their home cage, baseline bloodsamples were collected, and each animal received a single subcutaneous dose of 3 mg / kg LTW980.

[0384] Blood samples were collected at day -4 (baseline), day 0 (prior to dosing and 6 h post-dose), and days 1, 2, 3, 4, 7, 10, 14, 17, 21, 24, 28, 31, and 35 post-dose. All time points were timed from the end of administration of the dose given on day 0. Except for blood samples taken 6 h post-dose, samples were obtained in the morning prior to feeding, but the animals were not fasted overnight. Blood samples were collected cage-side from the saphenous vein into 3 mL syringes. One mL of blood was expelled into an EDTA-coated tube, 8.5 mg / mL aprotinin was added at 10 pL / mL of blood (final concentration = 0.085 mg / mL blood), and the tube was immediately placed on ice. Two mL of blood was expelled into a 3.5 mL serum separator tube and stored at RT for at least 30 min. Samples were centrifuged for 20 min (3,000 rpm at 4°C). Plasma / serum were divided into aliquots and stored at 80°C until analysis or shipping.

[0385] Human Fc IgG in cynomolgus monkey plasma was quantified using MesoScale Discovery electrochemiluminescent technology (MSD, Gaithersburg, MD) in a sandwich immunoassay. Anti-human-IgG mouse IgGl (R10Z8E9), a mouse monoclonal antibody against human IgG, was used as the capture antibody. MSD 96-well, MULTIARRAY single-spot plates were coated with 1 pg / mL anti-human-IgG mouse IgGl (30 pL / well) and incubated overnight at 4°C. The plates were then blocked with PBS / 5% BSA solution (150 pL / well) and incubated for 1 h. During the incubation period, LTW980 was serially diluted (300-0.03 ng / mL) in PBS / 5% BSA solution to obtain IgG standards for an 8- point standard curve and plasma samples were diluted 1 :30, 1 :300, and 1 :3000 in PBS / 5% BSA solution. Following the blocking period, the plates were washed 3 times with 400 pL IX PBS-T wash buffer using a BioTek EL406 automated plate washer (Winooski, VT), then diluted plasma samples and standards were immediately pipetted into the plates (25 pL / well). The plates were incubated for 2 h and then washed 3 times with 400 pL IX PBS-T wash buffer using the EL406 automated plate washer. MSD goat anti-human SULFO-TAG detection antibody (1 pg / mL) in PBS-T / 1% BSA was added to each well (25 pL / well) and the plates were incubated for 1 h. Plates were washed 3 times with 400 pL IX PBS-T wash buffer using the EL406 automated plate washer, then MSD IX read buffer T (150 pL / well) was pipetted into the plates. The plate electro-chemiluminescence was immediately detected using an MSD SECTOR Imager 6000 reader. LTW980 standards were assayed in triplicateper plate. Diluted samples were assayed in duplicate on separate plates. Curve fitting, back- calculation, % recovery, and interpolation of sample concentrations were performed using MSD DISCOVERY WORKBENCH Data Analysis Tools v4.0 Software. Signal generated by the IgG standards was plotted and fit using a 4-parameter logistical curve-fitting option with a l / y2 weighting function. Fc concentrations (ng / mL) in plasma samples were interpolated from the LTW980 standard curve and multiplied by dilution factors. The assay lower limit of quantification (LLOQ) was 0.9 ng / mL and the upper limit of quantification (ULOQ) was 900,000 ng / mL. LLOQ and ULOQ were defined as the lower and upper standard concentration with 100% recovery ± 20% and CV < 20% and then multiplied by the dilution factors.

[0386] Plasma triglyceride concentrations were measured using the Triglyceride (GPO) Liquid Reagent set (Pointe Scientific; Canton, MI; cat no. T7532-500). Pre-warmed assay reagent (300 pL, 37°C) was added to 5 pL of plasma in a clear, flat-bottom, 96-well plate (Thermo Fisher Scientific; Tewksbury, MA; cat no. 269620). The plate was mixed on a plate shaker for 30 s and then placed in an incubator at 37°C for 5 min. Following a 20 s mix, absorbance was measured at 500 nm with a SPECTRAmax PLUS plate reader. Triglyceride concentrations were calculated by comparing to a calibration curve generated using known quantities of a triglyceride standard (Pointe Scientific; cat no. T7531-STD).Safety assessments of LTW980 in monkeys

[0387] Cynomolgus monkeys' care was in accordance with institutional guidelines. The study protocols were reviewed and approved by the Institutional Animal Care and Use Committee (IACUC) of the facilities conducting the studies. Three toxicology studies were conducted with LTW980 in cynomolgus monkeys fed a low-fat or high-fat diet. In the first study, LTW980 (1, 10 and 100 mg / kg once weekly by subcutaneous injection) or vehicle was administered once weekly for 4 weeks to young, healthy monkeys fed a low-fat diet. In the second study, 7-year old male monkeys that were previously fed a high fructose, high fat diet as part of a separate metabolic assessment were fed a high-fat diet and dosed with vehicle or 30 mg / kg LTW980 subcutaneously, once weekly for 4 weeks. In the third study, young monkeys were fed a low-fat or high-fat diet and administered LTW980 by subcutaneous injection. In a third study, young healthy monkeys were fed a low- or high-fat diet, and LTW980 (0.3, 3, or 30 mg / kg for the high-fat diet groups; 30 mg / kg for the low-fatdiet group) or vehicle (low-fat and high-fat diet groups) was administered every 2 weeks for 15 weeks by subcutaneous injection. In the vehicle and 3 mg / kg high-fat diet groups, some animals comprised a recovery subgroup for which the terminal sacrifice occurred 16 weeks after the last dose. At necropsy, the MLN and surrounding intestinal tissue were embedded in a paraffin block, sectioned, and stained with hematoxylin and eosin and examined microscopically for the presence of foamy macrophages and any associated inflammation. Circulating SAA, NGAL, TEMPI, and CRP were assessed by immunoassays and 29 circulating cytokines, chemokines and growth factors were assessed using the Invitrogen Cytokine 29-Plex Monkey Panel.ResultsLTW980 lowered plasma triglycerides and remnant cholesterol in hypertriglyceridemic monkeys

[0388] In hypertriglyceridemic monkeys with baseline plasma triglyceride concentrations ranging from 207 to 2438 mg / dl, subcutaneous administration of a single dose of LTW980 (3 mg / kg) lowered mean plasma triglyceride levels by more than 40% from baseline within 3 days of dosing and produced a peak TG reduction of 58% on Day 7 post-dose (Table 5) On average mean plasma triglyceride levels were lowered by 41-58% on days 3-21 after dosing; triglyceride levels returned to baseline 28 days after dosing (Figure 13A). LTW980 plasma concentrations were above 2 pg / ml on days 3-21 after dosing (Figure 13B). The dose of 3 mg / kg LTW980 robustly lowered remnant cholesterol (RC), as shown in Table 6, which includes summary data for all four animals, and Figure 21, which depicts individual traces of an example animal, and Figure 37, which depicts individual traces for each of the three remaining animals tested (RC is shown as TRL in Figure 21). LTW980 robustly lowered TRL cholesterol levels (non-HDL-C, non-LDL-C) by 59% (Figure 13C). LTW980 decreased plasma total cholesterol by up to 28% and increased plasma HDL-C concentrations by up to 62%. LTW980 also reduced plasma apoC-III and apolipoprotein B concentrations by up to 28% and 30%, respectively (Figure 13D-E).

[0389] This example demonstrates that LTW980 is capable of lowering plasma triglycerides and remnant cholesterol in hypertriglyceridemic monkeys.Table 5. single dose of LTW980 (3 mg / kg) improves plasma lipid profile of HTG Cynomolgus monkeys.

[0390] To more accurately assess the effects of LTW980 on plasma lipoprotein profiles, we performed FPLC separation of plasma samples taken at baseline and Day 7 post-dose and measured cholesterol and TG concentrations in the fractions. LTW980 reduced TG content in the fractions containing TRL, LDL, and HDL particles consistently across all four animals dosed with LTW980 (Table 6, example animal shown in Figure 13F).Table 6. Lipoprotein triglyceride and cholesterol content on Day 7 post-dose derived fromFPLC analysis.

[0391] We also evaluated effects of LTW980 on lipoprotein cholesterol content using density gradient ultracentrifugation of plasma samples taken at baseline and Day 7 post-dose. Following LTW980 administration, the group displayed elevated HDLcholesterol content, no major change in LDL cholesterol content, and markedly lower TRL cholesterol content (Table 7).Table 7. Lipoprotein cholesterol content on Day 7 post-dose derived from density-gradient ultracentrifugation analysis.Safety assessment ofLTW980 in monkeys

[0392] In young healthy monkeys consuming a standard diet and administered LTW980 for 4 weeks, no pathology was observed. In a second study with older male monkeys fed a high-fat diet, weekly administration of LTW980 for 4 weeks resulted in foamy macrophage accumulation in the MLN (Figure 14) in 3 of the 6 LTW980-treated monkeys. One LTW980 -treated animal had local inflammation associated with some of the foamy macrophage aggregates (Figure 14). There was no evidence of systemic inflammation: no significant increase of circulating levels of the acute phase proteins SAA, hsCRP, or Fibrinogen (FIB) or of any of the proteins in a panel of 29 cytokines, chemokines and growth factors were observed in any of the LTW980-treated monkeys (Table 8.1 and Figure 38). No foamy macrophage accumulation was observed in monkeys fed a high-fat diet and treated with vehicle. In a third study in young healthy monkeys were fed a low- or high-fat diet and administered LTW980 or vehicle every two weeks for 15 weeks. In this study, minimal infiltrate of foamy macrophages in the MLN was observed in some of the animals fed a high-fat diet and treated with LTW980: in 1 of 3 males and 1 of 3 females given the 3 mg / kg / dose, and in each of 3 females and 0 of 3 males given the 30 mg / kg dose. This finding was most frequent in lymph nodes adjacent to the duodenum and was also noted in lymph nodes adjacent to the jejunum, ileum, cecum, and colon. There was no associated inflammation in the mesentery (assessed by histology) in this study, nor was there any evidence of a systemic inflammatory response: there was no change in circulating markers of inflammation (SAA, NGAL, TEMPI, CRP and a panel of 29 cytokines, chemokines and growth factors. No foamy macrophage infiltration was observed in themonkeys fed a low-fat diet or in monkeys fed a high-fat diet and treated with vehicle or the lowest dose of LTW980 (0.3 mg / kg / dose; Table 8).Table 8. Incidence and severity of LTW980-related histopathological changes in NHPs onHSFD with 15 weeks of treatment.able 8.1. Serum inflammatory biomarker levels in NHPs on HSFD administered LTW980 for 15 weeks.

[0393] In the LTW980-treated recovery group (3 mg / kg / dose) evaluated 16 weeks after the last dose (these animals continued to consume the high-fat diet during the recovery period), one female had persistence of minimal to slight foamy macrophage infiltrate in the MLN. Since the incidence of MLN findings was similar in the main phase and recovery 3 mg / kg LTW980 groups (25-33%), this study did not establish the reversibility of the foamy macrophage findings. Besides MLN foamy macrophage accumulation (and in one of the studies, associated local inflammation), no other safety-related findings were observed in the three monkey toxicology studies.

[0394] The potential for progression of the histological MLN findings in the 15- week study was assessed in a follow-on chronic dosing study with 9 months of LTW980 administration. In this second study, NHPs fed an HSFD received up to 30 mg / kg of LTW980 every 2 weeks for 36 weeks (20 total doses). Similar to the 15-week study, there were no LTW980-related effects on any in-life parameters, proinflammatory biomarkers, macroscopic examinations, or organ weights, and no adverse effects on clinical pathology examinations.

[0395] Consistent with the 15-week study, minimal to moderate FM formation in the MLN was present in a subset of treated animals with chronic LTW980 treatment (Table 9). Importantly, there was no worsening of the histological findings with chronic dosing, including no evidence of local MLN inflammation, degeneration, necrosis, fibrosis, or any other related changes in the mesentery. Similar to the 15-week study, there was no evidence of increased inflammatory biomarkers or clinical pathology in animals with histological findings.Table 9. Incidence and severity of LTW980-related histopathological changes in NHPs on HSFD with 36 weeks of treatment.

[0396] In summary, LTW980 reduced plasma triglycerides when administered to hypertriglyceridemic monkeys. In the monkey safety studies, the only findings were foamy macrophage aggregates in some MLN of some of the animals that were fed a high-fat diet; in one such animal there was local inflammation associated with the foamy macrophage aggregates. One of the monkey safety studies included a 16-week recovery group and did not establish the reversibility of the observed mesenteric lymphadenopathy.Example 3: Studies in mice

[0397] The present example demonstrates that foamy macrophage accumulation in the mesentery and associated local inflammation observed following administration of an anti-mouse ANGPTL4 antibody together with a high-fat diet were fully reversible after washout of anti-mouse ANGPTL4 antibody and transitioning the animals to a low-fat diet.The present study also demonstrates that mice treated with an anti-mouse ANGPTL4 antibody show reduced remnant cholesterol and improved insulin sensitivity (Figure 22).Material and methodsAssessment of plasma triglyceride lowering in 14D12 treated DBA / 2 mice

[0398] The anti-mouse ANGPTL4 antibody 14D12 was prepared by recombinant expression in suspension-adapted HEK cells followed by Protein A affinity chromatography. LTW980 is a humanized hlgGl-LALA / kappa monoclonal antibody. The LALA modification in the hinge region of the hlgGl Fc domain (i.e., Leu234 and Leu235 are replaced by Ala234 and Ala235) reduces the potential for antibody-dependent cellular cytotoxicity. Therefore, we prepared a 14D12 with a similar change in the m!gG2a Fc domain (i.e., Leu240 and Leu241 are replaced by Ala240 and Ala241).

[0399] Serum triglyceride concentrations were measured using a Wako TG kit with the following components: L-Type TG M Enzyme Color A (Rl), catalog no. 461-08992; L- Type TG M Enzyme Color B (R2), catalog no. 461-09092; Multi-Calibrator Lipid; catalog no. 464-01601; Control Serum I, catalog no. 410-00102; Control Serum II, catalog no. 416- 00202. Serum samples stored at -80 °C were thawed immediately prior to analysis, and 4-10 pL was added to wells of a 96-well plate (duplicate samples were prepared when sample volume was sufficient). Control serum I and II (4 pL) and each concentration (0, 13, 27, 47, 67, 88, and 101 mg / dL) of the Multi -Calibrator Lipid solutions (4-15 pL) were added to separate wells. PBS (0-11 pL) was then added to bring the total volume in each well to 15 pL. Enzyme Color A (Rl) solution (90 pL) was then added to each well, the contents of the wells were mixed by gentle rotation of the plate, and the plate was incubated at 37 °C for 5 minutes. Enzyme Color B (R2) solution (30 pL) was then added to each well, the plate was gently mixed as before, and incubated at 37 °C for 5 minutes. The absorbance of each well at 600 nm was then measured, and sample triglyceride concentrations were determined using a standard curve (absorbance vs. concentration) generated using the Multi-Lipid Calibrator.Assessment of reversibility of findings in the mesentery of mice treated with an ANGPTL4 inhibitor antibody

[0400] The care of mice used in this study was in accordance with institutional guidelines, and the study protocol was reviewed and approved by the Institutional AnimalCare and Use Committee (IACUC) of the facility conducting the study. DBA / 2 mice were fed a high saturated fat diet (45% fat in diet, D12451i, Research Diets) and administered the anti-mouse ANGPTL4 monoclonal antibody 14D12 (30 mg / kg) by intraperitoneal injection once weekly for 8 weeks. Except for one high-fat diet recovery group, the mice were transitioned to a low-fat diet following the last dose of 14D12. Groups of mice (n=6 / group) were sacrificed at the following timepoints after the last dose of 14D12: day 0, and 1, 7, 13, 19, 25, 31 and 37 weeks. The high-fat diet recovery group was sacrificed at 37 weeks. The presence and severity of foamy macrophage accumulation, foamy macrophage vacuolation, and associated inflammation in the mesentery and MLN were graded by assessing photomicrographs taken from hematoxylin- and eosin-stained tissue sections with microscope objectives of either 2x or 20x using Image Pro Plus (version 6.3.1.) and included the addition of scale bars.ResultsFoamy macrophage accumulation and inflammation in the mesentery of mice treated with an anti-mouse ANGPTL4 antibody is reversible

[0401] To induce foamy macrophage accumulation and associated local inflammation in the mesentery, DBA / 2 mice were fed a high saturated fat diet and treated for 8 weeks with an anti-mouse ANGPTL4 antibody, 14D1212 (30 mg / kg intravenous injection once per week). In these mice, 14D12 treatment lowered fasting plasma triglycerides by a similar extent in male and female mice (-64% in male mice and by -74% in female mice on study day 57; Figure 15, Table 10), which confirmed that mice were exposed to active concentrations of 14D12.Table 10. Mean serum triglyceride (TG) concentrations in DBA / 2 mice administered 14D12.Serum samples at each time point were collected from mice that were fasted for 4 to 5 hours prior to sample collection.aOnly 5 serum samples were available due to a technical issue with sample collection.*p < 0.05 vs. vehicle-treated group, t tests.

[0402] At the end of the dosing period, the following microscopic findings were observed in the lymphatic vessels in the mesentery (including the mesocolon) that drain to the MLN and in the MLN of males and females administered 14D12: foamy macrophage aggregates, vacuolation of the foamy macrophages, and associated mixed-cell inflammation (Figure 16, Figure 17, Table 11).Table 11. Incidence and severity of 14D12-related histopathological changes - main study, primary investigational groups.

[0403] No MLN enlargement was observed. 14D12-treated males had increased severity of the macrophage aggregates, as well as increased severity of cytoplasmic vacuolation of the foamy macrophages forming the aggregates when compared with females. This finding may be due to the larger quantity of the saturated fat diet ingested by male mice as compared with females. After cessation of treatment with 14D12 and switching the animals to a low-fat diet, the microscopic findings referred to above were completely reversed after 126 days of recovery in males, and after 84 days of recovery in females (Table 12, Table 13).Table 12. Incidence and severity of 14D12-related histopathological changes - Recovery phases, primary investigational groups - Males.aThe days of recovery was calculated as the number of days the mice were not dosed with 14D12 after the scheduled euthanasia day of the main study animals (i.e., recovery day 1 was study Day 57 and the first recovery necropsy was on study Day 99, which then had 42 days of recovery).bMale, number 2011, was found dead on study Day 79, following 22 days of recovery.c3 animals given reference item were necropsied at each time point, but all of these animals are reported here added together for simplicity in presenting the results.Table 13. Incidence and severity of 14D12-related histopathological changes - Recovery phases, primary investigational groups - Females.aThe days of recovery was calculated as the number of days the mice were not dosed with 14D12 after the scheduled euthanasia day of the main study animals (i.e., recovery day 1 was study Day 57 and the first recovery necropsy was on study Day 99, which then had 42 days of recovery).bF emale, number 1521, was found dead on study Day 256, following 199 days of recovery.c3 animals given reference item were necropsied at each time point, but all of these animals are reported here added together for simplicity in presenting the results.

[0404] In a subset of mice that continued to receive the high saturated fat diet during a 252-day recovery period, microscopic findings attributed to 14D12 treatment were partially reversed in male mice and fully reversed in female mice (Table 14).Table 14. Incidence and severity of 14D12-related histopathological changes - Recovery phase - High fat diet subset.All other microscopic findings, including the mixed cell inflammation and / or hemorrhage in the brown adipose tissue and mineralization in the heart (Percy and Barthold, 2007), were considered incidental, background in this strain and age of mouse and / or were of similar incidence and severity in reference item / control and treated mice and therefore were considered unrelated to the test item.

[0405] One 14D12-treated female mouse in the high saturated fat diet recovery group had microscopic findings (foamy macrophage aggregates, foamy macrophage vacuolation, mononuclear cell inflammation) in the mesentery that were focal and of low severity. Since similar findings were observed in one vehicle-treated male mouse, thesefindings were considered unlikely to be related to 14D12 treatment, but rather related to prolonged administration of the high saturated fat diet. In male mice, there were persistent microscopic findings (foamy macrophage aggregates, foamy macrophage vacuolation, mononuclear cell inflammation) in the mesentery of all 6 14D12-treated mice (Table 9), indicating a lack of reversibility of these findings. In the MLN, 3 out of 6 males had microscopic findings (foamy macrophage aggregates, foamy macrophage vacuolation, mononuclear cell inflammation) (Table 9), compared with 6 out of 6 at the end of the dosing phase, demonstrating a decrease in incidence, but not severity, of these findings after the 252-day recovery period, indicating partial recovery of these findings.

[0406] In summary, the mouse study showed that foamy macrophage accumulation in the mesentery and associated local inflammation observed following administration of an anti-mouse ANGPTL4 antibody (14D12) together with a high-fat diet were fully reversible after washout of 14D12 and transitioning the animals to a low-fat diet.Example 4: ANGPTL4 E40K homozygous human subjects

[0407] The present example demonstrates that humans who are homozygous carriers of an ANGPTL4 E40K loss-of-function variant do not have an increased rate of abdominal inflammatory disorders, suggesting that chronic ANGTL4 inhibition in humans does not lead to abdominal inflammatory disorders.

[0408] The E40K loss-of-function variant has been found to reduce ANGPTL4’s ability to inhibit LPL. E40K may abolish LPL inhibition by preventing extracellular accumulation N-terminal ANGPTL4 oligomers and E40K in vitro and in vivo functional experiments consistently show loss of 80% to 90% of LPL-inhibitory activity with ANGPTL4 E40K in comparison to a frameshift variant. ANGPTL4 inactivating variant has been robustly associated with reduced remnant cholesterol (RC), reduced TGs, improved insulin sensitivity, and concomitant protection from coronary artery disease (CAD) and T2D.Material and methods

[0409] To evaluate the impact of ANGPTL4 loss on subclinical MLN architecture, abdominal MRIs from 16 homozygous E40K carriers and 16 age, sex, and BMI-matchednoncarriers underwent independent, blinded radiological evaluation. A subset of three individuals who were homozygous carriers of the ANGPTL4 E40K variant (two men and one woman) were examined using either MRI or CT to evaluate potential mesenteric lymph nodes (MLN) pathology.Results

[0410] No lymph node enlargement or ascites were found in individuals with ANGPTL4 loss. Three individuals had MRI findings of mesenteric edema or fibrosis. Of these three individuals, who are homozygous carriers of the ANGPTL4 E40K variant, imaging of MLN by MRI or CT showed no evidence of MLN pathology: none had an MLN with a short axis greater than 6 mm or a chemical shift that might reflect MLN inflammation.Example 5: Safety, pharmacokinetics, triglyceride and remnant cholesterol lowering efficacy of anti-ANGPTL4 antibody LTW980 in human subjects

[0411] The present example demonstrates that anti-ANGPTL4 antibody (LTW980) was well tolerated and there was no evidence of inflammation as assessed by measurement of circulating inflammatory markers. The present example demonstrates that ANGPTL4 inhibition via administration of LTW980 can significantly lower plasma triglycerides and remnant cholesterol in human subjects with hypertriglyceridemia.Material and methodsStudy setup

[0412] LTW980 was evaluated in a first-in-human, randomized, subject-blinded, placebo-controlled, single ascending dose study. Informed consent was obtained after the nature and possible consequences of the studies had been explained. The trial was conducted at two clinical research organizations (CROs) in the United States. Institutional Board review approval was obtained at each participating site. Participants were recruited by the CROs using internal data bases as well as approved recruitment approaches. Written consent was obtained before any assessments were performed. The study design is outlined in Figure 1. The study included part IA, part IB, and part IC.• In part IA of the study, 4 dose escalation cohorts of healthy human subjects with normal triglyceride levels received LTW980 (15, 50, 150 or 450 mg) or placebo (for each cohort: n=6 LTW980, n=2 placebo) via subcutaneous injection.• In part IB of the study, a cohort of healthy human subjects with high body mass index (BMI 30 to 40 kg / m2) received a single 450 mg dose of LTW980 or placebo (n=9 LTW980, n=3 placebo) via subcutaneous injection.• In part IC of the study, a cohort of otherwise healthy human subjects with elevated fasting plasma triglycerides (200-500 mg / dL at the screening visit; note that in some human subjects fasting triglycerides were <200 mg / dl at the subsequent baseline visit) received a single 450 mg dose of LTW980 or placebo (n=8 LTW980, n=4 placebo).Eligible participants enrolled in Part IA were healthy men and women aged between 18 and 65, weighed at least 50 kg and had a BMI of between 18 and 30 kg / m2. Subjects in part IB weighed at least 70 kg and had a BMI within the range of 30-40 kg / m2. Randomization for parts IA and IB was 3: 1, LTW980 to placebo. Subjects in part IC weighed at least 59 kg and had triglycerides at the screening visit in the range of 200-500 mg / dl. Exclusion criteria included presence of peripheral lymphadenopathy, peritonitis within 5 years of the study, known hypersensitivity to any monoclonal antibody, significant illness that had not resolved with two weeks prior to initial dosing. Individuals with hepatitis B or C, malignancy of any organ system within the past five years or immunodeficiency disease were excluded. Pregnant or lactating women and women with child bearing potential were also excluded. For part IB and IC individuals taking medication aimed at lowering cholesterol, appetite or weight were excluded. Individuals with diabetes or a fasting blood glucose greater than 140 mg / dl were excluded, as were individuals with a history of abdominal disease such as pancreatitis, IBD, diverticulitis or pelvic inflammatory disease. Randomization in part IC was 2: 1 LTW980 to placebo.

[0413] The trial was designed as a single ascending dose trial to evaluate safety. A single dose of LTW980 was administered as summarized in Figure 1. Investigators and subjects were blinded as to treatment. Subjects were domiciled for 8 days after administration of LTW980. Safety was monitored by assessing adverse events, physicalexamination, local tolerability at the site of injection, vital signs, height and weight, laboratory evaluations (hematology, clinical chemistry, thyroid-stimulating hormone (TSH), urinalysis, fasting lipid panel), and electrocardiogram (ECG). hsCRP and SAA were measured in serum samples using an immunoassay. Inflammatory chemokines / cytokines (IL6, IL18, CXCL10 and TNFa) were measured in serum samples with an immunoassay on the Ella platform (Protein Simple). A standard glucose tolerance test was performed on the days noted. In brief, after an overnight fast, subjects consumed a solution containing 75 grams of glucose and serum was sampled prior to glucose administration and 30, 60 and 120 minutes after. For the meal tolerance test, after an overnight fast, subjects consumed a breakfast of 900 calories with 50-60% fat content. Serum samples were obtained before the meal and hourly for seven hours after the meal.

[0414] For all cohorts, the safety, tolerability, pharmacokinetics, and effect on fasting plasma triglyceride levels of LTW980 were assessed. LTW980 serum concentrations (mean ± standard deviation) following a single subcutaneous administration to human subjects for each cohort are shown in Figure 11. The effect of LTW980 on postprandial plasma triglyceride levels in response to a meal tolerance test was also assessed. All samples collected to evaluate efficacy and safety were analyzed at the local lab of the CRO conducting the study. Variability was addressed in statistical analysis. In part IC, serum inflammatory biomarkers (high-sensitivity CRP (hsCRP), serum amyloid A (SAA), haptoglobin, IL-6, IL-18, CXCL-10 and TNFa) were measured.ResultsSingle doses ofLTW980 lowered fasting and postprandial triglyceride levels in human subjects with elevated plasma triglyceride levels

[0415] Single subcutaneous (s.c.) injections of LTW980 or placebo were administered to healthy human subjects according to the study design shown in Figure 1. After establishing the safety and tolerability of increasing single doses of LTW980 in healthy human subjects, the highest tested dose of 450 mg s.c. was selected to further evaluate safety and efficacy in human subjects with an obese range BMI of over 30 kg / m2 with or without elevated triglyceride levels (part IB) and then in otherwise healthy human subjects with elevated baseline triglyceride levels (part IC). Subject demographics and baseline fasting triglyceride levels are listed in Table 15.Table 15. Clinical trial subject demographics and baseline fasting triglyceride levels.In the high triglyceride cohort (part IC), the mean baseline triglyceride level in LTW980-treated human subjects was 290 mg / dl (range: 179 to 527 mg / dl). In this cohort, LTW980 administration (450 mg s.c.) resulted in 42% and 47% lowering of mean fasting triglycerides compared to baseline levels on days 8 and 15 after dosing, respectively; triglyceride levels returned to baseline levels at 60 days post-dosing (Figure 2A, Table 16).Table 16. LTW980 administration (450 mg s.c.) lowered fasting triglyceride levels in human subjects with hypertriglyceridemia. (Analysis of ratio to baseline for fasting triglycerides in human subjects administered a single dose of LTW980 or placebo.Baseline is defined as the value at Day -1, 0 hrs pre-breakfast for triglycerides. Data were analyzed using an MMRM model including fixed effects for treatment and treatment* visit and log(baseline) as a covariate with an unstructured covariance matrix. The log(baseline)*visit interaction is also included in the MMRM model. One-sided p-values are reported due to the initial hypothesis that fasting triglycerides would be reduced after study drug administration.

[0416] In subjects from this cohort administered placebo, triglyceride levels did not decrease (Figure 4, Table 16). Placebo-adjusted mean triglyceride-lowering was calculated using a mixed model for repeated measures (MMRM) for days 8 and 15 after dosing (since maximal or near-maximal triglyceride lowering was observed on these days). The placebo- adjusted triglyceride lowering for part IC was 55% (1-sided p = 0.04) and 47% (1-sided p = 0.03) on days 8 and 15 after dosing, respectively (Table 17). LTW980 administration in this cohort also lowered the mean postprandial triglyceride excursion (AUCO-7 days) following a meal tolerance on day 5 after dosing test by 45% compared to baseline (Figure 3, Table 12) and by 57% compared to placebo (1-sided p = 0.0015; data were analyzed using an analysis of covariance (ANCOVA) model) (Table 12). Compared to placebo, the reduction was 72.5% (90% CI: -112.1 to -32.9%) (Table21). Single-dose LTW980 increased serum high-density lipoprotein cholesterol (HDL-C) by 13.5% (90% CI : 2.6 to 24.5%) compared to placebo on Day 15 and did not significantly change serum total cholesterol (TC) or LDL C levels (Figure 32).Table 17. LTW980 administration lowered postprandial triglycerides in subjects with hypertriglyceridemia. Analysis of ratio to baseline for postprandial triglycerides (AUC0-7hr) following a meal tolerance in subjects administered a single 450 mg dose of LTW980 or placebo.Baseline is defined as the value at Day -1. Data were analyzed using an ANCOVA model including fixed effect for treatment and log(baseline) as a covariate. One-sided p-values are presented due to the initial hypothesis that postprandial triglycerides would be reduced after study drug administration.

[0417] In the high triglyceride cohort (Part IC), mean remnant cholesterol at baseline was 47.2 mg / dL (range 31.9 to 66.8 mg / dL) in the LTW980 group and 31.6 (range23.6 to 36.5 mg / dL) in the placebo group. The LTW980-treated subjects showed a 56% placebo-adjusted reduction in RC from baseline to Day 15 (Figure 20).

[0418] The high triglyceride cohort (Part 1 C) furthermore demonstrated a significant improvement in TG / HDL ratio at both Days 15 and 43 (Figure 24), which has been linked to an improvement in insulin resistance. Further, fasting TG reductions were evident as early as Day 2 and reached maximal levels on Day 15 (Figure 30) and free LTW980 PK parameters are summarized in Table 24 Placebo-adjusted TG and RC reduction (90% CI) was -68.6% (- 97.4 to -39.8%) and -65.6% (-94.2 to -36.9) on Day 15, respectively (Figure 20). Drug exposure was maintained for over 40 days, as evidenced by persistent TG and RC lowering, and returned to baseline levels by Day 57. In the high-TG cohort, 450 mg LTW980 also reduced the mean postprandial TG excursion (AUC 0-7 days) by 42.7% (90% CI: -48.6 to - 36.8%) compared to baseline in a meal tolerance test conducted on Day 5 after dosing (Figure 3). Compared to placebo, the reduction was 72.5% (90% CI: -112.1 to -32.9%) (Table 21). Single-dose LTW980 increased serum high-density lipoprotein cholesterol (HDL-C) by 13.5% (90% CI : 2.6 to 24.5%) compared to placebo on Day 15, and did not significantly change serum total cholesterol (TC) or LDL-C levels (Figures 32A-32C). In Parts IA and IB, baseline fasting TG levels were normal (69 and 146 mg / dL, respectively). Accordingly, 450 mg of LTW980 provided less robust fasting TG reductions, and did not lower postprandial TG (Table 20 & 21).

[0419] Triglyceride-lowering was also assessed in parts IA-4 and IB of the study (Figure 5 and 6). In these cohorts, baseline fasting triglyceride levels were lower than in part IC: mean baseline triglyceride levels in LTW980-treated subjects in cohorts IA-4 and LB were 69 mg / dl and 146 mg / dl, respectively (Table 10). In these cohorts, LTW980 administration (450 mg s.c.) resulted in less robust placebo-adjusted mean triglyceride- lowering compared to that observed in part IC: statistically significant triglyceride-lowering (1-sided p < 0.05) was observed for only one of the two timepoints (day 8 or day 15 after dosing) for which p-values were calculated (Table 16). In parts IA-4 and IB, administration of 450 mg LTW980 did not lower postprandial triglycerides (Table 17). In Part IC, LTW980 administration did not significantly increase plasma LDL cholesterol or HDL cholesterol levels (although a trend to increased HDL-C was observed) (Figure 7, Figure 8), and did not affect glucose or insulin levels during an oral glucose tolerance test conducted on day 8 after dosing (Figure 9). Serum inflammatory biomarkers (hsCRP, SAA, haptoglobin, IL-6, IL-18,CXCL-10 and TNFa) were measured in Part IC and no elevations of these markers were observed in subjects treated with LTW980 (Figure 10 and Figure 33).

[0420] In all parts of the study, the LTW980 plasma concentration versus time profiles for part showed more rapid clearance of the antibody at later time points, which is indicative of target-mediated drug disposition (Figure 2B, Figure 16). No serious adverse events were reported in LTW980-treated subjects. Reported adverse events were mild in severity with no observable pattern across the cohorts (Table 18 and 22, Table 19 and 23).Table 18. Adverse events (AEs) observed in part I A.Table 20: Change in triglyceride levels on Day 15 after single dose L:TW980 administration.Triglyceride levels at baseline are represented as Mean ± SD. Change from baseline and placebo-adjusted percent change from baseline represented as mean change (90% CI)Table 21: Change in postprandial triglycerides (AUC0-7hr) following a meal tolerance test.Change from baseline and placebo-adjusted percent from baseline represented as mean change (90% CI)N = number of participants studied, nE = number of AE events in the category, nS = number of participants with at least one AE in the category.% is based on number of participantsN = number of participants studied, nE = number of AE events in the category, nS = number of participants with at least one AE in the category.Table 24: Summary statistics of LTW980 serum PK parametersExample 6: Safety and efficacy of anti-ANGPTL4 monoclonal antibody in patients with familial partial lipodystrophy / FPLD

[0421] The present example will demonstrate proof of concept efficacy in patients with Familial Partial Lipodystrophy (FPLD).

[0422] Familial partial lipodystrophy syndromes (FPL) are rare and highly morbid diseases. These disorders are characterized by selective loss of adipose tissue with insulin resistance and its concomitant metabolic complications including diabetes, dyslipidemia and hepatic steatosis. Patients are at extremely elevated risk for atherosclerotic cardiovascular disease, recurrent acute pancreatitis, cirrhosis, as well as significant impairments in quality of life and patient satisfaction.

[0423] Treatment of patients with FPLD with anti-ANGPTL4 monoclonal antibody LTW980 may improve circulating biomarkers (including triglycerides and fasting insulin) and may reduce liver fat. These effects may reflect clinically meaningful improvements in lipodystrophy and may have a potential to substantially reduce the risk of cardiometabolic diseases such as diabetes, NASH, and atherosclerotic cardiovascular disease (ASCVD).Material and methodsStudy setup

[0424] One or more doses of LTW980 will be evaluated in FPLD / FPLD-adjacent human cohorts. The study design is outlined in Figures 18 and 19. The study will include part IIA, part IIB, and part IIC. The inclusion criteria are listed below.Inclusion criteria for part IIA (FPLD-adjacent):1) Age > 18 and < 75 years;2) hypertriglyceridemia (>250 mg / dL);3) FPLD-like body composition defined by DXA showing FMR > >1.2 for women and > 1.7 for men (Screen with WHR-adjBMI); and4) Fasting insulin > 10 pIU / mL

[0425] Inclusion criteria for part IIB and IIC (FPLD):1) Age > 18 and < 75 years;2) Hypertriglyceridemia (>250 mg / dL);3) Clinical diagnosis of lipodystrophy defined by either a or b: a. Presence of known FPLD disease-causing variant or b. Fat loss as objectively quantified by meeting at least one of :■ decreased midthigh skin fold thickness■ imaging showing symmetric, selective absence of fat from the gluteofemoral region (FMR > 1.3 in women and FMR > 2 in men)4) and at least one of the metabolic conditions■ Severe insulin resistance (fasting insulin > 10 pIU / mL)■ Diagnosis of type 2 diabetes mellitus■ Nonalcoholic fatty liver disease (clinical diagnosis or liver fat fraction)5) Women of childbearing potential agreeing to use highly effective method of contraception for the duration of the trial and duration after.

[0426] The following exclusion criteria will be used:1. Acute or chronic severe liver disease as evidenced by any of the following: encephalopathy, variceal bleeding, INR > 1.7 in the absence of anti coagulation treatment2. Terminal illness with expected survival of less than 1 year3. Presence of peripheral lymphadenopathy on physical exam. Also, any history of mesenteric disease (including lymphadenitis) or peritonitis within the last 5 years4. Hypersensitivity to any monoclonal antibody5. Enrollment or planned enrollment in another therapeutic clinical trial in next 3 months or within 5 half-lives of last prior dose of another experimental therapeutic6. Currently taking metreleptin (Myalept), or have received metrereleptin in the 3 months prior to signing informed consent.7. Inability to comply with planned study procedures8. Pregnancy or breastfeeding mothers9. History of HIV, hepatitis B or C infection10. History of acquired lipodystrophy11. History of familial Chylomicronemia or familial hypertryglicerademia (not lipodystrophy)12. History of Type 1 Diabetes13. Cancer diagnosis within 1 year excluding skin cancer or carcinoma in situ of the cervix14. History of PCI or coronary bypass within past 6 months15. BMI > 3516. History of Celiac disease or inflammatory bowel disease17. Current smoking or active recreational drug use18. Positive Hepatitis B surface antigen or Hepatitis C test result.19. History of immunodeficiency diseases, including a positive HIV (ELISA andWestern blot) test result.20. Active disease of any of the following in the 6 months prior to baseline: cholelithiasis, inflammatory bowel disease, irritable bowel syndrome, diverticulitis, pelvic inflammatory disease, prostatic disease or disease of the endometrium or ovary.21. History of abdominal surgery within 6 months of screening, or any other surgical procedure within 10 weeks of screening.22. Consuming very low-carbohydrate, high fat diets (subject self-reports as being on a ketogenic, “paleo”, or Atkins style diet)23. Not being able to undergo MRI (Details to be provided)24. Any condition which in the opinion of the investigator prevents the subject from complying with study procedures or prevents the subject from completing the study or interferes with the interpretation of study results

[0427] Two trials will be designed, trial 1 and trial 2.Trial 1 - dose finding study:

[0428] Four cohorts of subjects having FPLD-adj acent will receive LTW980 (50, 150 or 450 mg) or placebo (for each cohort: n=20 LTW980, n=20 placebo) via subcutaneous injections. Subjects will have an enrolment period of two weeks (Week 2) and thereafter biweekly receive LTW980 injections for 14 weeks (dosing at the beginning of each week: Week 4 through Week 16 of the trial), hereby receiving a total of 7 doses of LTW980. Investigators and subjects will be blinded as to treatment. Blood samples will be obtained at baseline (Week 2) and every second week through week 16 (Week 4 to Week 16). An imaging will be performed on the days noted in Figure 18 (after Week 2, Week 8 and Week 16 of Trial 1).

[0429] Optionally, if no signal / effect is observed after 3 doses, subjects can be rerandomized into another dose cohort.Trial 2 - proof of concept of LTW 980 in the FPLD cohorts:

[0430] This is a randomized, parallel group, double-blind, placebo-controlled study in patients with FPLD. The study will include a Screening period (4-week), single-blind placebo run-in, and a 12- week treatment period and safety follow-up.

[0431] Eligible patients will receive a single-blind injection of placebo on Day 1 and Day 15 in a 4-week run-in period in an attempt to establish a stable baseline of key biomarkers. Patients will be randomized at the beginning of Week 4 to one of 2 doses of LTW980 (150 or 450 mg) or placebo (for each cohort: n=7 LTW980, n=7 placebo) via subcutaneous injections. Dosing is illustrated in the Figure 19. After the enrolment period of four weeks (Week 4) the patients will biweekly receive LTW980 subcutaneous injections for 12 weeks (dosing at the beginning of each week: Week 4 through Week 16 of the trial), hereby receiving a total of 7 doses of LTW980. LTW980 and a placebo developed to match LTW980 will be provided in syringes or vials. Investigators and subjects will be blinded as to treatment. Blood samples will be obtained at baseline (fist 4 weeks) and every second week through week 16 (Week 4 to Week 16). An imaging will be performed after Week 2, Week 8 and Week 16 of Trial 1. Optionally, if sufficient efficacy is observed after 3 doses, subjects can be moved from the 450 mg LTW980 dose cohort into the 150 mg LTW980 dose cohort. Patients will be instructed not to change their dietary habits during the run-in period or during the study (Trial 2).Endpoints

[0432] The effect of LTW980 on fasting triglyceride levels, liver fat fraction (measured by MRI), incidence and frequency of treatment-emergent adverse events compared to baseline levels will be assessed. The effect of LTW980 on postprandial plasma triglyceride levels in response to a meal tolerance test will be assessed. The effect of LTW980 on systemic inflammatory markers, such as hsCRP, SAA, IL-6, IL- 19, CXCL-10, and TNF-a, and mesenteric lymphadenopathy (measured by MRI) compared to baseline levels will be assessed.

[0433] Furthermore, the effect of LTW980 on insulin resistance (including fasting insulin levels and HOMA-IR), glucose homeostasis (fasting glucose, postprandial glucose and hemoglobin ale), diabetes medication (changes in insulin dose, and / or changes in other hypoglycemic drug doses), lipid metabolism (HDL / LDL / VLDL levels and AUC on meal tolerance test), body fat distribution (fat mass ratio measured by DEXA and / or MRI), ectopic fat distribution (midthigh skin fold thickness), AdipoIR, gluteofemoral fat (using stable isotope tracers and fat biopsies), body composition (measured by MRI quantified fat in gluteofemoral, abdominal subcutaneous, visceral depots,), skeletal and muscle fat, serum leptin and adiponectin levels (total and adjusted for BMI), and serum insulin level will be assessed.

[0434] The pharmacokinetic (PK) profile of LTW980 in FPLD patients will be assessed by• Maximum observed concentration (Cmax),• Time to maximum concentration (tmax),• Area under the curve overdosing interval (AUC(O-tau))• Clearance (CL),• Volume of distribution (Vz),• Terminal half-life (tl / 2)• Concentration at the end of dose interval (Ctrough)Example 7: Safety and efficacy of anti-ANGPTL4 monoclonal antibody in patients with metabolic dysfunction

[0435] The present example will demonstrate proof of concept efficacy in patients with metabolic dysfunction.

[0436] Metabolic dysfunction is a constellation of multiple interrelated risk factors (e.g., including, but not limited to, increased triglycerides (TG), waist circumference, fasting plasma glucose (FPG) and hypertension) that combine to increase metabolic disease burden in the form of risk for type 2 diabetes and atherosclerotic cardiovascular disease (ASCVD). Patients with metabolic dysfunction are also at risk of related metabolic associated diseases such as non-alcoholic steatohepatitis (NASH) and heart failure with preserved ejection fraction (HFpEF), among others.

[0437] As described in Example 6, familial partial lipodystrophy syndromes (FPL) are rare and highly morbid diseases. There is significant overlap in terms of the clinical manifestations of metabolic dysfunction and FPLD with the exception of the stereotypical changes in body composition and the selective loss of gluteofemoral adipose tissue in FPLD.

[0438] Lipoprotein lipase (LPL) is an enzyme that plays a key role in clearing TG from the bloodstream, by hydrolyzing triglycerides (TG) carried by TG-rich lipoproteins (TRL). LPL activity is regulated by multiple mechanisms including natural protein activators and inhibitors such as apolipoprotein C-II (apoC-II) and apolipoprotein C-III (apoC-III), respectively. ANGPTL4 is an endogenous inhibitor of LPL and may therefore be an attractive potential target for pharmacologic inhibition.

[0439] LTW980 is an ANGPTL4 antibody that binds to the N-terminal CCD of ANGPTL4 and prevents ANGPTL4-mediates inhibition of LPL.

[0440] Since there is an overlap in the clinical manifestations of patients with metabolic dysfunction and FPLD, we plan to enroll subjects with a subset of the components of metabolic dysfunction that most closely mimic the clinical manifestations of FPLD.

[0441] Treatment of patients with metabolic dysfunction with anti-ANGPTL4 monoclonal antibody LTW980 may improve metabolic dysfunction markers including triglycerides, fasting glucose, fasting insulin and / or will reduce liver fat. These effects may reflect clinically meaningful improvements that may have mechanistic similarities to what we may observe in patients with FPLD treated with LTW980. Therefore, this study in patients with metabolic dysfunction should not only offer important insight for future studies in metabolic dysfunction patients, but also offer insights for future studies in FPLD.Material and methodsStudy setup

[0442] Proof of mechanism on insulin resistance of multiple doses of LTW980 in a metabolic dysfunction human cohort will be evaluated after 6 weeks of treatment.Characterizing safety and tolerability of multiple doses of LTW980 in a metabolic dysfunction human cohort will be evaluated after 12 weeks of treatment.Inclusion criteria for Trial 3 (patients with metabolic dysfunction):1) Age > 18 and < 75 years;2) hypertriglyceridemia (>250 mg / dL);3) Severe insulin resistance (fasting insulin > 30plU / mL);4) Nonalcoholic fatty liver disease (clinical diagnosis or AST: ALT <1 plus evidence of steatosis on a liver ultrasound);5) Women of childbearing potential agreeing to use highly effective method of contraception for the duration of the trial and a duration after.The following exclusion criteria will be used:1. Acute or chronic severe liver disease as evidenced by any of the following: encephalopathy, variceal bleeding, INR > 1.7 in the absence of anti coagulation treatment2. Terminal illness with expected survival of less than 1 year3. Presence of peripheral lymphadenopathy on physical exam. Also, any history of mesenteric disease (including lymphadenitis) or peritonitis within the last 5 years4. Hypersensitivity to any monoclonal antibody5. Enrollment or planned enrollment in another therapeutic clinical trial in next3 months or within 5 half-lives of last prior dose of another experimental therapeutic6. Inability to comply with planned study procedures7. Pregnancy or breastfeeding mothers8. History of HIV, hepatitis B or C infection9. History of familial Chylomicronemia or familial hypertryglicerademia10. History of Type 1 Diabetes11. History of Type 2 diabetes on any diabetes medication other than metformin12. Cancer diagnosis within 1 year excluding skin cancer or carcinoma in situ of the cervix13. History of PCI or coronary bypass within past 6 months14. BMI > 3515. History of Celiac disease or inflammatory bowel disease16. Current smoking (>10 cigarettes per day) or active recreational drug use other than marijuana / THC17. A history of hepatitis B or hepatitis C or a Positive Hepatitis B surface antigen or Hepatitis C test result at screening.18. History of immunodeficiency diseases, including a positive HIV (ELISA and Western blot) test result.19. Active disease of any of the following in the 6 months prior to baseline: cholelithiasis, inflammatory bowel disease, diverticulitis, pelvic inflammatory disease, or disease of the endometrium.20. History of abdominal surgery within 6 months of screening, or any other surgical procedure within 10 weeks of screening.21. Consuming very low-carbohydrate, high fat diets (subject self-reports as being on a ketogenic, “paleo”, or Atkins style diet)22. Not being able to undergo MRI23. Any condition which in the opinion of the investigator prevents the subject from complying with study procedures or prevents the subject from completing the study or interferes with the interpretation of study results24. Current use of >2 alcoholic beverages per dayTrial 3 - proof of concept of LTW980 in patients with metabolic dysfunction:

[0443] This is a randomized, parallel group, double-blind, placebo-controlled study in patients with metabolic dysfunction. Approximately 60 subjects having metabolic dysfunction will be enrolled and will complete a single-blind placebo run-in, with the goal to randomizing at least 15 subjects per treatment group. The study will include a Screening period (4-week), single-blind placebo run-in (4 weeks), and a 12- week treatment period and an 8 week safety follow-up.

[0444] Eligible patients will receive a single-blind injection of placebo once a week during the 4-week run-in period in an attempt to establish a stable baseline of key biomarkers due to dietary and other behavioral changes associated with enrolment in a clinical trial. Patients will be randomized at the beginning of Week 5 (after the placebo run- in) to one of 3 doses of LTW980 (300 mg weekly, 150 mg weekly or 150 mg bi-weekly) or placebo: 15 will be randomized to placebo, 15 to LTW980 300 mg weekly, 15 to LTW980 150 mg weekly and 15 to LTW980 150 mg bi-weekly.

[0445] Active LTW980 and a matching placebo will be provided in vials containing ImL of solution. Each active viral contains 150 mg / mL of LTW980. Patients will receive subcutaneous (SC) injection weekly or biweekly. Patients will be instructed not to change their dietary habits during the run-in period or during the study.Dosing Justification

[0446] Pharmacokinetics / pharmacodynamics (PK / PD) data derived from the three dose levels may provide adequate data for exposure-response analyses, which in turn would inform the dose to be used in the pivotal study.

[0447] A visual inspection of the plasma concentration vs time and the percent reduction in TG vs time curves from the Phase 1 study indicated a lag in achieving the nadir for TG relative to Tmax, which in turn resulted in a hysteresis in the plasma concentration vs effect plot. A simple Emax model with an effect compartment was used to assess the relationship between plasma LTW980 concentration and change in serum triglyceride from the cohort of subjects with hypertriglyceridemia in the Phase 1 study. The effect compartment served to collapse the hysteresis.

[0448] This model adequately fitted the observed data from subjects with elevated serum triglycerides (290 mg / dL at baseline) treated with a single 450 mg subcutaneous (SC) dose. Based on the model, 150 mg once-weekly SC injection of LTW980 may provide near- maximal decrease in fasting serum triglyceride (46% decrease; mean fasting serum triglyceride of -155 mg / dL at steady-state) and is associated with a Cmaxof 45 ug / mL and of 38 ug / mL at steady-state. 150 mg weekly (QW) is considered an appropriate dose and is the mid-dose for the proposed study. The high dose is selected as twice the 150mg QW dose, i.e., 300 mg QW while the low dose is selected to be half the 150 mg QW dose, i.e., 150 mg biweekly (Q2W). The choice of 150 mg Q2W dose rather than 75 mg QW doseis based on the sub-linear pharmacokinetics observed in the Phase 1 study over 15mg to 50 mg and 75 mg being too close to that range. The 4-fold dose range afforded by the selected doses, coupled with inter-subject variability, should provide roughly 8-fold range in plasma concentrations, which in turn would be adequate to describe the exposure-response relationship from the proposed Phase 2 study.

[0449] The projected exposures at the highest proposed dose of 300 mg weekly (Cmax of 89 ug / mL; AUCo-tau of 13,860 ug / mL.hr) are below the exposures associated with no adverse effect level (NOAEL) in the 15-week study in cynomolgus monkeys (Cmax = 954 ug / mL; AUCo-i4d = 250,000 ug / mL.hr).Endpoints

[0450] The effect of LTW980 on insulin resistance (fasting insulin and fasting glucose), fasting triglyceride levels, liver fat fraction (measured by e.g., MRI), systemic inflammatory markers (high-sensitivity C-reactive protein (hsCRP), serum amyloid A (SAA), IL-6, IL-19, CXCL-10, and TNF-a), diabetes medication dosage in metabolic dysfunction patients (changes in insulin dose and other hypoglycemic drugs’ dose), and mesenteric lymphadenopathy (measured by MRI) compared to baseline levels will be determined. Safety and tolerability will be characterized by comparing incidence and frequency of treatment-emergent adverse events, changes from baseline in clinical laboratory parameters, vital signs and EKG parameters. The effect of LTW980 on postprandial plasma triglyceride levels in response to a meal tolerance test will be assessed.

[0451] Furthermore, the effect of LTW980 on insulin resistance (including fasting insulin levels and HOMA-IR), insulin / glucose homeostasis (fasting glucose, postprandial glucose and hemoglobin ale), liver fat fraction, triglyceride metabolism, lipid metabolism (HDL / LDL / VLDL levels and AUC on meal tolerance test), body fat distribution (change in visceral fat mass and abdominal fat mass), and ectopic fat distribution (pericardial fat mass, % skeletal fat mass, and / or pancreatic fat mass measured by MRI) will be determined.

[0452] The effect of LTW980 on sex hormone biomarkers in pre-menopausal women will be determined.

[0453] The pharmacokinetic (PK) profile of LTW980 in metabolic dysfunction patients will be assessed by• Maximum observed concentration (Cmax)• Time to maximum concentration (tmax)• Area under the curve overdosing interval (AUC(O-tau))• Clearance (CL)• Volume of distribution (Vz)• Terminal half-life (tl / 2)• Concentration at the end of dose interval (Ctrough)• Formation of anti-LTW980-antibodiesExample 8: Safety and efficacy of anti-ANGPTL4 antibody in humans with Atherosclerotic cardiovascular disease (ASCVD)

[0454] The present example will demonstrate proof of concept efficacy in patients with atherosclerotic cardiovascular disease (ASCVD).

[0455] ASCVD may be caused by plaque buildup in arterial walls and may include, but is not limited to, diseases that comprise coronary heart disease (CHD), such as myocardial infarction, angina, and coronary artery stenosis; cerebrovascular disease, such as a transient ischemic attack, ischemic stroke, and carotid artery stenosis; peripheral artery disease, such as claudication; or aortic atherosclerotic disease, such as abdominal aortic aneurysm and descending thoracic aneurysm. Currently, ASCVD- related conditions remain a leading cause of morbidity and mortality globally.

[0456] Treatment of patients with ASCVD with an anti-ANGPTL4 monoclonal antibody described herein (LTW980) may improve ASCVD markers including remnant cholesterol levels, triglycerol, LDL, HbAlc, Insulin resistance, fasting inclusion and fasting glucose levels, liver fat and inflammatory markers.Material and methodsStudy setup

[0457] One or more doses of LTW980 will be evaluated in an ASCVD human cohort. The study is a randomized, parallel group, double-blind, placebo-controlled study in patients with ASCVD. Following screening, participants enter 24 week dosing and 12-week follow-up periods. Patients will receive subcutaneous (SC) injection weekly, biweekly prmonthly. Patients will be instructed not to change their dietary habits during the run-in period or during the study.Inclusion criteria (patients with ASCVD):1) high risk of cardiovascular events;2) history of ASCVD; and / or3) remnant cholesterol >40 mg / dL.Proof of concept ofLTW980 in patients with ASCVD:

[0458] Proof of mechanism on remnant cholesterol levels, LDL, HbAlc, Insulin resistance, fasting inclusion and fasting glucose levels, liver fat and inflammatory markers of multiple doses of LTW980 in a ASCVD human cohort will be evaluated after treatment with anti-ANGPTL4 monoclonal antibody LTW980.Endpoints

[0459] Key endpoints are pharmacokinetics, safety (including MRI assessments of mesenteric lymph nodes) and tolerability, and RC and TG levels. Exploratory endpoints include effects on insulin resistance, MRI to evaluate ectopic liver and other tissue fat, and fasting glucose and HbAlc.Example 9: Anti-ANGPTL4 Antibody, LTW-980, trial for multi-dose safety and tolerability

[0460] The present example demonstrates multi-dose safety and tolerability in patients with metabolic dysfunction in a Phase 2a trial.

[0461] As described in Example 7, metabolic dysfunction is a constellation of multiple interrelated risk factors (e.g., including, but not limited to, increased triglycerides (TG), waist circumference, fasting plasma glucose (FPG) and hypertension) that combine to increase metabolic disease burden in the form of risk for type 2 diabetes and atherosclerotic cardiovascular disease (ASCVD). Patients with metabolic dysfunction are also at risk of related metabolic associated diseases such as non-alcoholic steatohepatitis (NASH) and heart failure with preserved ejection fraction (HFpEF), among others.

[0462] LTW980 is an ANGPTL4 antibody that binds to the N-terminal CCD of ANGPTL4 and prevents ANGPTL4-mediates inhibition of LPL.

[0463] Preclinical non-human primate (NHP) studies were performed for toxicology and safety. A 9-month Good Laboratory Practices (GLP)-compliant showed that NHPs administered LTW980 biweekly in subcutaneous (SC) injections had no effects on any inlife parameters (e.g., cardiovascular, respiratory, or CNS), clinical pathology, inflammatory biomarkers, macroscopic examination, or organ weights. Analysis of safety and toxicology measures demonstrated that No-Observable-Adverse-Effect-Level (NOAEL) were detected at the highest tested doses (~7-8x higher than dosing in Phase 2b trial described in Example 10).

[0464] Key objectives of the Phase 2a trial were to: (1) establish safety and tolerability at effective doses, and (2) demonstrate lowering of triglyceride (TG) and remnant cholesterol (RC) levels as well as a PK / PD relationship. Subgroup analysis was performed in patients with TG >150 mg / dL and >200 mg / dL at baseline (Table 26 and 27). No drug-related safety or tolerability signals were detected. Treatment of metabolic dysfunction patients with anti-ANGPTL4 monoclonal antibody LTW980 improved metabolic dysfunction markers including TG and RC as described below.

[0465] Adults with metabolic dysfunction were screened at baseline for age, BMI, type 1 diabetes mellitus, hemoglobin Ale (HbAlc), TG, RC, and LDL cholesterol (Table 25). Remnant cholesterol was calculated by the Sampson equation (See Sampson et. al. Biomedicines 2022, 10, 3156, which is hereby incorporated by reference in its entirety).Table 25. Baseline characteristics of participants in Phase 2a trial.Material and methodsPhase 2a Study setup

[0466] LTW980 doses of 150 mg, 300 mg, and 450 mg were evaluated to establish safety and tolerability as well as efficacy. Following screening, adult participants with TG levels greater that 150mg / dL (predictive of fasting RC levels greater than 30 mg / dL) entered a two-week placebo run-in, where all participants were started on placebo. Following the two-week run-in, participants entered a 12-week dosing and 12-week follow-up period. During the 12-week dosing period, patients received biweekly (Q2W) subcutaneous (SC) injections of 150 mg (n = 10), 300 mg (n=9), 450 mg (n=17), or placebo (n=19) for 12 weeks followed by a 12-week follow up period.Results

[0467] Topline results showed no SAEs, AEs, or drug-related safety and tolerability signals. Additionally, no drug-related increases in inflammatory markers were detected. LTW980 doses of 300 mg and 450 mg substantially lowered TG and RC levels versus placebo in participants who had TG levels lower than 150 mg / dL at baseline (Figure 25 A- B).

[0468] Dose-dependent increases in the serum concentrations of LTW980 at 7 days after administration of the first dose (Week 1) were observed. In addition, trough concentrations at Week 12 increased relative to those observed in Week 1 in each dose cohort (Figure 39). LTW980 doses of 300 mg and 450 mg doses substantially lowered TG and RC levels versus placebo at 12 weeks in participants who had triglyceride levels lower than 150 mg / dL at baseline (Figure 26A-B). In the 150 mg, 300 mg, and 450 mg treatment groups, the week 12 placebo-adjusted percent TG lowering calculated for each group were 32.6, 42.4, and 44.7, respectively (Figure 26A). In the 150 mg, 300 mg, and 450 mg treatment groups the placebo-adjusted percent of RC lowering calculated at week 12 were 30.2, 42.1, and 45.3, respectively (Figure 26B).

[0469] LTW980 doses of 300 mg and 450 mg doses substantially lowered TG and RC levels versus placebo at 12 weeks in participants who had TG levels greater than 150 mg / dL at baseline (Figure 27A-B). In the 150 mg, 300 mg, and 450 mg treatment groups, the placebo-adjusted percent TG lowering calculated for each group at week 12 relative to baseline were 35.8, 52.1, and 53.0, respectively (Figure 27A). In the 150 mg, 300 mg, and 450 mg treatment groups the placebo-adjusted percent RC lowering calculated for eachgroup for each group at week 12 relative to baseline at week 12 were 32.3, 51.4, 54.0, respectively (Figure 27B).

[0470] Subgroup analysis showed greater efficacy in participants with higher baseline triglycerides (Tables 26 and 27).Table 26. Subgroup analysis of triglyceride and remnant cholesterol level changes in the participants with baseline triglyceride levels greater than 150 mg / dL.Table 27. Subgroup analysis of triglyceride and remnant cholesterol level changes in the participants with baseline triglyceride levels greater than 200 mg / dL

[0471] At Week 12, LTW980 treatment resulted in significant and dose-dependent reductions in TG and RC (Figure 31 and Figure 34). Analysis of individual participants showed that administration of LTW980 at 300 mg and 450 mg doses resulted in consistent TG and RC lowering (% change from baseline at 12 weeks; Figure 28A and 28B, respectively).

[0472] These Phase 2a results demonstrated clinical efficacy as -50% TG and RC lowering predicts a meaningful clinical benefit compared to -20-35% relative risk reduction (RRR) in major adverse cardiovascular events (MACE) predicted by ANGPTL4 human genetics and RC epidemiology (See Elias Bjbmson et al. European Heart Journal, October2023, which is hereby incorporated by reference in its entirety). Together, LTW980 450 mg treatment resulted in a placebo-adjusted mean percent change (90% CI) in TG of -64.0% (- 101.5 to -26.6% ) in participants with baseline and >200 mg / dL, respectively (Table 28). Similarly, LTW980 450 mg resulted in placebo-adjusted mean percent changes (90% CI) in RC of -66.0% (-103.9 to -28.0%) in participants with baseline TG >200 mg / dL (Table 23).Table 28. Change in RC at Week 12 stratified by baseline TG levels in multidose study.

[0473] In addition to reductions in TG and RC levels, we observed treatment-related reductions in non-HDL-C and VLDL-C, and an increase in HDL-C at Week 12 (Figure 34). LTW980 450 mg produced placebo-adjusted mean reductions (90% CI) of 52.3% (-76.2 to - 28.3%) in VLDL-C, 7.1% (-14.7 to -0.6%) in non-HDL-C, and 19.0% (10.2 to 27.8%) increase in HDL-C. There were no treatment-related changes in TC or LDL-C.

[0474] Overall, LTW980 treatment was safe and well-tolerated. The proportions of participants with treatment-emergent AEs (TEAEs) was balanced across LTW980 treatment groups and placebo (Table 29). There were no SAEs or AEs associated with death. Only 1 participant (placebo group) in the study discontinued study treatment due to an AE. The majority of AEs were Grade 1 (mild; 22 [40.2%] participants) or Grade 2 (moderate; 24 [43.6%] participants). One participant in the placebo group experienced both a Grade 3 and a Grade 4 AE, which were high cholesterol and hypertriglyceridemia, respectively. Overall, 21 (38.2%) participants experienced an AE which were considered likely related to study treatment, with no clear trend across treatment groups (Table 29).Table 29. Overview of AEs in the multidose clinical study

[0475] Based on the observation of mesenteric lymphadenitis in ANGPTL4 knockout mice fed a HSFD, enhanced monitoring was used to assess safety related to the mesentery. Events of abdominal pain and gastrointestinal (GI) AEs or symptoms such as nausea, vomiting, diarrhea, and constipation, were predefined as AEs of special interest (AESIs) and closely monitored. In addition, systemic inflammatory markers, including hsCRP, SAA, and fibrinogen were measured. Finally, participants underwent abdominal MRI at Baseline, Week 12 (end of treatment), and Week 24 (end of study) to assess changes from baseline in MLN size, fat content, and local inflammation.

[0476] In total 15 (27.3%) participants experienced an AESI: 5 (26.3%) in the placebo, 2 (20.0%) in the LTW980 150 mg, 2 (22.2%) in the LTW980 300 mg, and 6 (35.3%) in the LTW980 450 mg groups (Table 30). Overall, the most common AESI was nausea in 6 (10.9%) participants. In total, 7 (12.7%) participants experienced AESIs that were considered related to treatment: 1 (5.3%), 3 (30.0%), and 3 (33.3%) participants in the placebo, 150 mg, and 300 mg groups respectively; none occurred in the 450 mg LTW980 group. No persistent or treatment-related increases in inflammatory biomarkers occurred in any treatment group (Figure 35).Table 30. AESI observed in multidose clinical study

[0477] There were no clinically meaningful differences in standard biochemical safety measures, and no worsening of metabolic parameters related to LTW980 treatment, including no worsening of insulin resistance, as measured by fasting glucose, fasting insulin, and homeostatic model assessment of insulin resistance (HOMA-IR; Figure 36).Furthermore, there were no increases in glycated hemoglobin levels (HbAlc). Lastly, there was no worsening of liver fat fraction measured by MRI proton density fat fraction (MRL PDFF).

[0478] The MRI evaluation revealed no LTW980 treatment-related changes in MLN size or number, and no imbalance in the appearance of newly detected nodes between baseline and Week 12 MRI scans in any LTW980 dose group compared with placebo (Table 27). At Week 12, of 77 nodes assessed in the placebo group, 49.4% either stayed the same or decreased whereas 50.6% increased in size; of 50 nodes assessed in the 450 mg group, 52.0% decreased in size and 48.0% increased. There were only 3 clinically relevant nodes (defined as >10 mm25) observed from baseline to Week 12: 2 occurred in the placebo group and 1 occurred in the 450 mg LTW980 group (Table 32). No treatment-related fat content increase occurred in the node observed in the LTW980 450 mg treated participant.Table 31. Overview of safety biomarkers in multidose clinical stud...

Claims

1. CLAIMSWe claim:

1. A method of treating a human having a disorder comprising: subcutaneously administering a dose of about 15 mg to about 1000 mg of an antibody or antigen-binding fragment thereof that binds specifically to human angiopoietin- like 4 protein (ANGPTL4) about weekly to about annually to the human, thereby improving at least one sign or symptom of the disorder in the human after administration.

2. The method of claim 1, wherein the dose is about 150 mg to about 450 mg.

3. The method of claim 2, wherein the dose is about 150 mg, about 225 mg, about 300 mg, or about 450 mg.

4. The method of claim 3, wherein the dose is about 225 mg or about 450 mg.

5. The method of any one of claims 1-4, wherein the dose is administered about weekly, about biweekly, about monthly, about quarterly, about biannually, or about annually.

6. The method of any one of claims 1-4, wherein the dose is administered about every 2 weeks, about every 3 weeks, about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, or about every 12 weeks.

7. The method of claim 6, wherein the dose is administered about every 4 weeks or about every 8 weeks.

8. The method of any one of claims 1-7, wherein the human is afflicted with a cardiovascular disease.

9. The method of claim 8, wherein the cardiovascular disease comprises atherosclerotic cardiovascular disease (ASCVD), a myocardial infarction (MI), stroke, coronary revascularization, arteriosclerosis, or any combination thereof.

10. The method of claim 9, wherein the cardiovascular disease comprises ASCVD.

11. The method of any one of claims 1-10, wherein the disorder is obesity, fatty liver disease, insulin resistance, metabolic dysfunction, metabolic syndrome, polycystic ovarian syndrome, chronic kidney disease, Cushing syndrome, hypercorti soli sm, acromegaly, heart failure, lipodystrophy, diabetes, or any combination thereof.

12. The method of any one of claims 1-11, wherein the human is afflicted with one or more of hyperlipidemia, hypercholesterolemia, hyperlipoproteinemia, hypertriglyceridemia, and dyslipidemia.

13. The method of any one of claims 1-12, wherein the human is afflicted with an elevated fasting plasma triglyceride concentration of at least about 150 mg / dl.

14. The method of any one of claims 1-13, wherein the human is afflicted with an elevated fasting plasma insulin concentration of at least about 6 plU / ml.

15. The method of any one of claims 1-14, wherein the antibody or antigen -binding fragment thereof is administered in combination with a second therapy.

16. The method of any one of claims 1-15, wherein one or more of fasting plasma triglyceride levels, fasting plasma remnant cholesterol levels, fasting plasma HbAlc levels, fasting plasma insulin levels, and fasting plasma glucose levels are measured prior to administration of one or more doses.

17. The method of any one of claims 1-15, wherein one or more of fasting plasma triglyceride levels, fasting plasma remnant cholesterol levels, fasting plasma HbAlc levels, fasting plasma insulin levels, or fasting plasma glucose levels are measured at least about 1 day after administering one or more doses.

18. The method of any one of claims 1-17, wherein the step of administering the antibody or antigen-binding fragment thereof results in a reduction in one or more of plasma triglyceride levels, plasma remnant cholesterol levels, plasma HbAlc levels, plasma insulin levels, and plasma glucose levels compared to basal levels.

19. The method of any one of claims 1-18, wherein the antibody or antigen-binding fragment thereof comprises:(a) a heavy chain variable region (VH) comprising a HCDR1 of SEQ ID NO: 7, a HCDR2 of SEQ ID NO: 8, and a HCDR3 of SEQ ID NO: 9, and a light chain variable region (VL) comprising a LCDR1 of SEQ ID NO: 17, a LCDR2 of SEQ ID NO: 18, and a LCDR3 of SEQ ID NO: 19; or(b) a VH comprising a HCDR1 of SEQ ID NO: 10, a HCDR2 of SEQ ID NO: 11, and a HCDR3 of SEQ ID NO: 12, and a VL comprising a LCDR1 of SEQ ID NO: 20, a LCDR2 of SEQ ID NO: 21, and a LCDR3 of SEQ ID NO: 22.

20. The method of claim 19, wherein the antibody or antigen-binding fragment thereof is a monoclonal antibody, humanized antibody, single chain antibody, Fab fragment, Fv fragment, F(ab')2 fragment, or scFv fragment.

21. The method of claim 19 or 20, wherein the antibody or antigen-binding fragment is an IgGl or IgG4 isotype.

22. The method of any one of claims 19-21, wherein the antibody or antigen-binding fragment comprises:(i) a VH comprising an amino acid sequence that is at least 90% identical to SEQ ID NO: 13, and(ii) a VL comprising an amino acid sequence that is at least 90% identical to SEQ ID NO: 23.

23. The method of claim 22, wherein the antibody or antigen-binding fragment comprises:(i) a VH comprising an amino acid sequence of SEQ ID NO: 13, and(i) a VL comprising an amino acid sequence of SEQ ID NO: 23.

24. The method of claim 23, wherein the antibody comprises:(i) a heavy chain comprising an amino acid sequence that is at least about 90% identical to SEQ ID NO: 28, SEQ ID NO: 156, SEQ ID NO: 157, or SEQ ID NO: 15, and(ii) a light chain comprising an amino acid sequence that is at least about 90% identical to SEQ ID NO: 25.

25. The method of claim 24, wherein the antibody comprises:(i) a heavy chain comprising an amino acid sequence of SEQ ID NO: 28, SEQ ID NO: 156, SEQ ID NO: 157, or SEQ ID NO: 15, and(ii) a light chain comprising an amino acid sequence of SEQ ID NO: 25.

26. The method of claim 25, wherein the antibody comprises:(i) a heavy chain comprising an amino acid sequence of SEQ ID NO: 28, and(ii) a light chain comprising an amino acid sequence of SEQ ID NO: 25.

27. The method of claim 25, wherein the antibody comprises:(i) a heavy chain comprising an amino acid sequence of SEQ ID NO: 15, and(ii) a light chain comprising an amino acid sequence of SEQ ID NO: 25.

28. The method of claim 25, wherein the antibody comprises:(i) a heavy chain comprising an amino acid sequence of SEQ ID NO: 156, and(ii) a light chain comprising an amino acid sequence of SEQ ID NO: 25.

29. The method of claim 25, wherein the antibody comprises:(i) a heavy chain comprising an amino acid sequence of SEQ ID NO: 157, and(ii) a light chain comprising an amino acid sequence of SEQ ID NO: 25.

30. The method of any one of claims 1-29, wherein the antibody or antigen-binding fragment thereof is formulated in a pharmaceutical composition comprising a pharmaceutically acceptable carrier.