Methods for treating leukocytosis, endothelial dysfunction, and carditis using lipid-binding protein-based conjugates - Patents.com

Abstract

A method of treating leukocytosis, endothelial dysfunction, and carditis comprising administering to a subject a lipid-binding protein-based conjugate.

Description

[Technical field]

[0001] 1. CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 328,210, filed April 6, 2022, the contents of which are incorporated herein by reference in their entirety. [Background technology]

[0002] 2. Sequence Listing This application contains a Sequence Listing that has been submitted electronically and is incorporated herein by reference in its entirety. Said electronic copy, created on Apr. 3, 2023, is an XML file with the filename CRN-048WO_ST26 and is 3,244 bytes in size. Summary of the Invention

[0003] 3. Summary The present disclosure is based, in part, on the discovery that subjects treated with the lipid-binding protein-based conjugate CER-001 described in Example 2 showed unexpectedly rapid improvements in biomarkers of inflammation, leukocytosis, and endothelial dysfunction.

[0004] Thus, in some aspects, the present disclosure provides methods of treating a subject having or at risk of developing leukocytosis with a predetermined dose (e.g., a high dose) of a lipid-binding protein-based conjugate.

[0005] In some aspects, the present disclosure provides a method of treating a subject having one or more symptoms associated with leukocytosis using a dose (e.g., a high dose) of a lipid-binding protein-based conjugate, comprising administering to the subject a dose of the lipid-binding protein-based conjugate.

[0006] In some aspects, the present disclosure provides methods of treating a subject with endothelial dysfunction (e.g., a subject having or who has experienced an acute coronary syndrome or stroke) with a dose (e.g., a high dose) of a lipid-binding protein-based complex.

[0007] In some aspects, the present disclosure provides methods of treating a subject experiencing or having experienced an acute coronary syndrome or stroke with high doses of lipid-binding protein-based complexes.

[0008] In some aspects, the disclosure provides methods of treating a subject having or at risk of developing carditis with a dose (e.g., a high dose) of a lipid-binding protein-based conjugate.

[0009] In the methods of the present disclosure, the lipid-binding protein-based complex is typically administered in a high dose. The high dose is typically higher than the dose that would be used to treat a chronic condition such as familial hypercholesterolemia. The high dose is typically administered over a relatively short period of time, for example over a period of 2 days to 2 weeks, and typically includes multiple administrations of the lipid-binding protein-based complex, for example 2-20 individual doses. The individual doses may be separated by less than a day (e.g., two administrations per day) or by more than a day (e.g., one administration per day).

[0010] In some embodiments of the disclosed method, the lipid-binding protein-based complex comprises sphingomyelin and / or negatively charged lipids, such as CER-001. CER-001 is a negatively charged lipoprotein complex that comprises recombinant human ApoA-I, sphingomyelin (SM), and 1,2-dihexadecanoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (dipalmitoylphosphatidyl-glycerol, DPPG). It mimics natural nascent discoid pre-beta HDL, the form that HDL particles take before acquiring cholesterol. Without being bound by theory, it is believed that CER-001 treatment can (1) reduce serum levels of inflammatory cytokines such as IL-6, (2) reduce white blood cell counts in subjects with or at risk for developing leukocytosis, and (3) reduce serum levels of ICAM-1 and VCAM-1, thereby providing clinical benefit to subjects with or at risk for developing leukocytosis, subjects with endothelial dysfunction, and subjects with or at risk for developing carditis.

[0011] In some aspects, the disclosure provides dosing regimens of lipid binding protein-based therapy (e.g., CER-001 therapy) for a subject described herein.

[0012] The dosing regimen of the present disclosure typically involves multiple administrations of CER-001 to the subject (e.g., daily administration or administrations approximately 12 hours apart). CER-001 treatment can be continued for a predetermined period of time, e.g., one week or longer than one week (e.g., two weeks). Alternatively, administration of CER-001 to the subject can be continued until one or more symptoms of the condition experienced by the subject have decreased, or until serum levels of one or more inflammatory markers have decreased, e.g., decreased to normal levels or decreased compared to baseline measurements taken before the initiation of CER-001 treatment. In subjects with an infection, in some embodiments, treatment can be continued until the subject has recovered from the infection.

[0013] The dosing regimens of the present disclosure may involve administering a lipid-binding protein-based conjugate (e.g., CER-001) to a subject according to an initial "induction" regimen, optionally followed by administering a lipid-binding protein-based conjugate to a subject according to a "consolidation" regimen.

[0014] An induction regimen typically involves administering multiple doses of a lipid-binding protein-based conjugate (eg, CER-001) to the subject, for example, six doses over a three day period.

[0015] The consolidation therapy regimen typically comprises administering one or more doses of lipid-binding protein-based conjugate (e.g., CER-001) to the subject after the final dose of the induction regimen, for example, one or more days after the final dose of the induction regimen. In some embodiments, the first dose of the consolidation therapy regimen is administered on the third day after the final dose of the induction regimen. For example, the dosing regimen can comprise administering lipid-binding protein-based conjugate (e.g., CER-001) to the subject following the induction regimen on days 1, 2, and 3, and administering lipid-binding protein-based conjugate to the subject following the consolidation therapy regimen on day 6. In some embodiments, the consolidation therapy regimen comprises two doses of lipid-binding protein-based conjugate.

[0016] In certain embodiments, the present disclosure provides a method of treating a subject described herein with a lipid binding protein-based conjugate (e.g., CER-001) according to a dosing regimen comprising: - Two doses per day on days 1, 2, and 3 (induction regimen), optionally followed by - Two subsequent doses on day 4 and beyond (consolidation regimen). In some embodiments, the regimen includes: - Two doses per day on days 1, 2, and 3 (induction regimen), followed by - 2 doses on day 6 (consolidation regimen).

[0017] In some embodiments, the dosing regimen comprises a single phase, such as one that corresponds to the induction regimen described herein.

[0018] In certain embodiments, the lipid-binding protein-based conjugate (eg, CER-001) is administered in combination with standard of care, such as antibiotic treatment, for infection and / or hemodynamic support.

[0019] In certain embodiments, an antihistamine (e.g., dexchlorphenylamine, hydroxyzine, diphenhydramine, cetirizine, fexofenadine, or loratadine) can be administered prior to administration of the lipid-binding protein-based conjugate (e.g., CER-001). The antihistamine can reduce the likelihood of an allergic reaction. 4. Brief description of the drawings [Brief description of the drawings]

[0020] [Figure 1] FIG. 1 shows IL-6 serum levels in a porcine model of sepsis-induced AKI (Example 1). [Diagram 2] FIG. 2 shows soluble VCAM-1 serum levels in a porcine model of sepsis-induced AKI (Example 2). [Diagram 3] FIG. 2 shows soluble ICAM-1 serum levels in a porcine model of sepsis-induced AKI (Example 3). [Figure 4] FIG. 1 shows LPS serum levels in a porcine model of sepsis-induced AKI (Example 1). [Diagram 5] FIG. 2 shows a schematic diagram of the clinical study of Example 2. [Figure 6] FIG. 1 shows a flow sheet of the Example 3 study. [Figure 7] FIG. 1 shows a flow sheet for the Example 4 study. [Figure 8-1] In the first cohort of subjects treated in the study of Example 2, MCP1 (FIG. 8A), TNF-α (FIG. 8B), [Figure 8-2] VCAM (Figure 8C), ICAM (Figure 8D), [Figure 8-3] Ferritin (Figure 8E), leukocytes (Figure 8F), [Figure 8-4] CRP (Fig. 8G), KIM-1 (Fig. 8H), [Figure 8-5] FIG. 8I shows changes in IL-8 (FIG. 8I), and triglycerides (FIG. 8J). [Figure 9-1] 9A-9B show changes in VCAM in the standard of care (SOC) group and the three CER-001 groups in the clinical study of Example 2. FIG. 9A, Change from baseline for the SOC group versus the aggregate CER-001 group. FIG. 9B, Change from baseline for each group. [Figure 9-2] Figure 9C, Change as percentage of peak for SOC group and pooled study group. Figure 9D, Change as percentage of peak for SOC group and study group broken down by whether subjects were enrolled from the center's ICU or nephrology department. [Figure 9-3] Figure 9E, Change from baseline for each subject in the SOC and aggregate CER-001 groups. Figure 9F, Change from baseline for each subject in the SOC group and each study group. [Figure 10-1] Figure 10 shows the change in ICAM for the standard of care (SOC) group and the three CER-001 groups in the clinical study of Example 2. Figure 10A, change from baseline for the SOC group versus the aggregate CER-001 group. Figure 10B, change from baseline for each group. [Figure 10-2] Figure 10C, Change as percentage of peak for SOC group and pooled study group. Figure 10D, Change as percentage of peak for SOC group and study group broken down by whether subjects were enrolled from the ICU or nephrology department of the center. [Figure 10-3] Figure 10E, Change from baseline for each subject in the SOC and aggregate CER-001 groups. Figure 10F, Change from baseline for each subject in the SOC group and each study group. [Figure 11]FIG. 1 shows the results of an MTT cell viability assay of cultured endothelial cells upon challenge with LPS and CER-001 infusions as described in Example 6. [Figure 12] FIG. 11 summarizes the results of endothelial nitric oxide synthase (eNOS)-based (eNOS (phospho-S1177)) FACS of cultured endothelial cells upon challenge with LPS and CER-001 infusions as described in Example 6. [Figure 13] FIG. 1 shows the results of FACS based on eNOS (phospho-S1177) of cultured endothelial cells compared to basal and VEFG (positive control) cells in one representative of three independent experiments upon challenge with LPS and CER-001 infusion as described in Example 6. [Figure 14] FIG. 1 shows the results of an MTT cell viability assay of PBMC from healthy donors upon challenge with LPS and CER-001 infusions as described in Example 6. [Figure 15] FIG. 13 shows the synthesis of TNF-α in PBMC from healthy donors upon challenge with LPS and CER-001 infusions as described in Example 6. [Figure 16] FIG. 1 shows the results of CD14-based FACS of PBMC from healthy donors in one representative of three independent experiments upon challenge with LPS and CER-001 infusion as described in Example 6. [Figure 17] FIG. 11 summarizes the results of CD14-based FACS of PBMC from healthy donors upon challenge with LPS and CER-001 infusion as described in Example 6. [Figure 18-1] 18A and 18B show the change in white blood cell counts for the standard of care (SOC) and CER-001 groups in the clinical study of Example 2. FIG. 18A shows the change in white blood cell counts from baseline for the SOC and aggregate CER-001 groups. FIG. 18B shows the change in white blood cell counts from baseline for the SOC and respective CER-001 groups. [Figure 18-2]Figure 18C: Change in white blood cell counts for the SOC and aggregate CER-001 groups reported as a percentage of peak white blood cell counts (peak=100%). The effect of treatment x study day on peak was p=0.5492. Figure 18D: Change in white blood cell counts for the SOC and aggregate CER-001 groups broken down by whether subjects were enrolled from the ICU or nephrology department of the center. [Figure 18-3] Figure 18E: Individual data points summarized in Figure 18A. Figure 18F: Individual data points summarized in Figure 18B. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0021] 5. Detailed Description In some aspects, the disclosure provides methods of treating a subject having or at risk of developing leukocytosis with a predetermined dose (e.g., a high dose) of a lipid-binding protein-based conjugate.

[0022] In some aspects, the present disclosure provides a method of treating a subject having one or more symptoms associated with leukocytosis using a dose (e.g., a high dose) of a lipid-binding protein-based conjugate, comprising administering to the subject a dose of the lipid-binding protein-based conjugate.

[0023] In some aspects, the present disclosure provides methods of treating a subject with endothelial dysfunction (e.g., a subject having or who has experienced an acute coronary syndrome or stroke) with a dose (e.g., a high dose) of a lipid-binding protein-based complex.

[0024] In some aspects, the present disclosure provides methods of treating a subject experiencing or having experienced an acute coronary syndrome or stroke with high doses of lipid-binding protein-based complexes.

[0025] In some aspects, the disclosure provides methods of treating a subject having or at risk of developing carditis with a dose (e.g., a high dose) of a lipid-binding protein-based conjugate.

[0026] In some embodiments of the method of the present disclosure, the lipid-binding protein-based complex is Apomer, Cargomer, HDL-based complex or HDL mimic-based complex.In a specific embodiment, the lipid-binding protein-based complex is CER-001.

[0027] Exemplary features of lipid-binding protein-based complexes that can be used in the methods and compositions of the present disclosure are described in Section 5.1. Exemplary subject populations that can be treated with the compositions of the present disclosure by the methods of the present disclosure are described in Section 5.2.

[0028] In some embodiments, the method of the present disclosure comprises administering a lipid-binding protein-based conjugate (e.g., CER-001) to a subject in two stages. First, the lipid-binding protein-based conjugate (e.g., CER-001) is administered in an initial, high-intensity "induction" regimen. The induction regimen is followed by a less intense "consolidation therapy" regimen. Alternatively, the lipid-binding protein-based conjugate (e.g., CER-001) can be administered to a subject in a single stage, for example, according to a dosing regimen that corresponds to the dose and dosing frequency of the induction or consolidation therapy regimen described herein.

[0029] Induction regimens that can be used in the methods of the present disclosure are described in Section 5.3, and consolidation therapy regimens that can be used in the methods of the present disclosure are described in Section 5.3.2. The dosing regimens of the present disclosure include administering a lipid-binding protein-based complex (e.g., CER-001) as a monotherapy or as part of a combination therapy with one or more agents, for example, in combination with standard treatments for sepsis or other infections, such as antibiotic treatment and / or hemodynamic support. Combination therapy is described in Section 5.4.

[0030] 5.1. Lipid-binding protein-based complexes 5.1.1. Complexes based on HDL and HDL mimetics In one embodiment, the lipid-binding protein-based complex comprises a complex based on HDL or HDL mimic.For example, the complex can comprise a lipoprotein complex described in US Pat. No. 8,206,750, WO 2012 / 109162, WO 2015 / 173633 (e.g. CER-001), or US Patent Publication No. 2004 / 0229794, the contents of each of which are incorporated herein by reference in their entirety.The terms "lipoprotein" and "apolipoprotein" are used interchangeably herein, and the term "lipoprotein" encompasses lipoprotein mimics, unless otherwise required by the context.The terms "lipid-binding protein" and "lipid-binding polypeptide" are also used interchangeably herein, and these terms do not imply a specific length of amino acid sequence, unless otherwise required by the context.

[0031] Lipoprotein complexes can include a protein fraction (e.g., an apolipoprotein fraction) and a lipid fraction (e.g., a phospholipid fraction). The protein fraction can include one or more lipid-binding protein molecules, such as apolipoproteins, peptides, or peptide analogs or mimetics of apolipoproteins, e.g., one or more lipid-binding protein molecules described in Section 5.1.2.

[0032] The lipid fraction typically includes one or more phospholipids, which may be neutral, negatively charged, positively charged, or a combination thereof. Exemplary phospholipids and other amphipathic molecules that may be included in the lipid fraction are described in Section 5.1.4.

[0033] In certain embodiments, the lipid fraction contains at least one neutral phospholipid (e.g., sphingomyelin (SM)) and, optionally, one or more negatively charged phospholipids. In lipoprotein complexes that include both neutral and negatively charged phospholipids, the neutral and negatively charged phospholipids can have fatty acid chains with the same or different numbers of carbons and the same or different degrees of saturation. In some examples, the neutral and negatively charged phospholipids have the same acyl tail, e.g., C16:0 or palmitoyl acyl chains. In specific embodiments, particularly those using egg SM as the neutral lipid, the weight ratio of apolipoprotein fraction:lipid fraction ranges from about 1:2.7 to about 1:3 (e.g., 1:2.7).

[0034] Any phospholipid that has at least a partial negative charge at physiological pH can be used as the negatively charged phospholipid. Non-limiting examples include phosphatidylinositol, phosphatidylserine, phosphatidylglycerol, and the negatively charged forms, such as salts, of phosphatidic acid. In a specific embodiment, the negatively charged phospholipid is 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)], or DPPG, phosphatidylglycerol. Preferred salts include potassium and sodium salts.

[0035] In some embodiments, the lipoprotein complexes used in the methods of the present disclosure are lipoprotein complexes described in U.S. Pat. No. 8,206,750 or WO 2012 / 109162 (and its U.S. counterpart, U.S. Patent Application Publication No. 2012 / 0232005), the contents of each of which are incorporated herein by reference in their entireties. In certain embodiments, the protein components of the lipoprotein complex are described in Section 6.1 and preferably Section 6.1.1 of WO 2012 / 109162 (and US 2012 / 0232005), and the lipid components are described in Section 6.2 of WO 2012 / 109162 (and US 2012 / 0232005), which may optionally be complexed together in amounts described in Section 6.3 of WO 2012 / 109162 (and US 2012 / 0232005). The contents of each of these sections are incorporated herein by reference. In certain embodiments, the lipoprotein complexes of the present disclosure are in a population of complexes that is at least 85%, at least 90%, at least 95%, at least 97%, or at least 99% homogeneous, as described in Section 6.4 of WO 2012 / 109162 (and U.S. Patent Application Publication No. 2012 / 0232005), the contents of which are incorporated herein by reference.

[0036] In a specific embodiment, a lipoprotein complex that can be used in the methods of the present disclosure comprises 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 50-80 lecithin molecules, and 20-50 SM molecules.

[0037] In another specific embodiment, a lipoprotein complex that can be used in the methods of the present disclosure comprises 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 50 lecithin molecules, and 50 SM molecules.

[0038] In yet another specific embodiment, a lipoprotein complex that can be used in the methods of the present disclosure contains 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 80 lecithin molecules, and 20 SM molecules.

[0039] In yet another specific embodiment, a lipoprotein complex that can be used in the methods of the present disclosure contains 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 70 lecithin molecules, and 30 SM molecules.

[0040] In yet another specific embodiment, a lipoprotein complex that can be used in the methods of the present disclosure contains 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 60 lecithin molecules, and 40 SM molecules.

[0041] In a specific embodiment, a lipoprotein complex that can be used in the methods of the present disclosure consists essentially of 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 50-80 lecithin molecules, and 20-50 SM molecules.

[0042] In another specific embodiment, a lipoprotein complex that can be used in the methods of the present disclosure consists essentially of 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 50 lecithin molecules, and 50 SM molecules.

[0043] In yet another specific embodiment, a lipoprotein complex that can be used in the methods of the present disclosure consists essentially of 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 80 lecithin molecules, and 20 SM molecules.

[0044] In yet another specific embodiment, a lipoprotein complex that can be used in the methods of the present disclosure consists essentially of 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 70 lecithin molecules, and 30 SM molecules.

[0045] In yet another specific embodiment, a lipoprotein complex that can be used in the methods of the present disclosure consists essentially of 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 60 lecithin molecules, and 40 SM molecules.

[0046] In specific embodiments, lipoprotein complexes that can be used in the methods of the present disclosure comprise a lipid component that includes about 90-99.8 wt% SM and about 0.2-10 wt% negatively charged phospholipids, e.g., about 0.2-1 wt%, 0.2-2 wt%, 0.2-3 wt%, 0.2-4 wt%, 0.2-5 wt%, 0.2-6 wt%, 0.2-7 wt%, 0.2-8 wt%, 0.2-9 wt%, or 0.2-10 wt% total negatively charged phospholipids. In another specific embodiment, a lipoprotein complex that can be used in the methods of the present disclosure comprises about 90-99.8 wt% lecithin and about 0.2-10 wt% negatively charged phospholipids, for example, about 0.2-1 wt%, 0.2-2 wt%, 0.2-3 wt%, 0.2-4 wt%, 0.2-5 wt%, 0.2-6 wt%, 0.2-7 wt%, 0.2-8 wt%, 0.2-9 wt%, or 0.2-10 wt% total negatively charged phospholipids.

[0047] In specific embodiments, lipoprotein complexes that can be used in the methods of the present disclosure comprise a lipid component consisting essentially of about 90-99.8 wt% SM and about 0.2-10 wt% negatively charged phospholipids, e.g., about 0.2-1 wt%, 0.2-2 wt%, 0.2-3 wt%, 0.2-4 wt%, 0.2-5 wt%, 0.2-6 wt%, 0.2-7 wt%, 0.2-8 wt%, 0.2-9 wt%, or 0.2-10 wt% total negatively charged phospholipids. In another specific embodiment, the lipoprotein complex that can be used in the methods of the present disclosure consists essentially of about 90-99.8 wt% lecithin and about 0.2-10 wt% negatively charged phospholipids, e.g., about 0.2-1 wt%, 0.2-2 wt%, 0.2-3 wt%, 0.2-4 wt%, 0.2-5 wt%, 0.2-6 wt%, 0.2-7 wt%, 0.2-8 wt%, 0.2-9 wt%, or 0.2-10 wt% total negatively charged phospholipids.

[0048] In yet another specific embodiment, the lipoprotein complex that can be used in the methods of the present disclosure comprises a lipid fraction containing about 9.8-90 wt% SM, about 9.8-90 wt% lecithin, and about 0.2-10 wt% negatively charged phospholipids, for example, from about 0.2-1 wt%, 0.2-2 wt%, 0.2-3 wt%, 0.2-4 wt%, 0.2-5 wt%, 0.2-6 wt%, 0.2-7 wt%, 0.2-8 wt%, 0.2-9 wt%, 0.2-10 wt% total negatively charged phospholipids.

[0049] In yet another specific embodiment, the lipoprotein complex that can be used in the methods of the present disclosure comprises a lipid fraction consisting essentially of about 9.8-90 wt% SM, about 9.8-90 wt% lecithin, and about 0.2-10 wt% negatively charged phospholipids, for example, from about 0.2-1 wt%, to 0.2-2 wt%, 0.2-3 wt%, 0.2-4 wt%, 0.2-5 wt%, 0.2-6 wt%, 0.2-7 wt%, 0.2-8 wt%, 0.2-9 wt%, 0.2-10 wt% total negatively charged phospholipids.

[0050] In another specific embodiment, a lipoprotein complex that can be used in the methods of the present disclosure comprises an ApoA-I apolipoprotein and a lipid fraction, the lipid fraction comprising sphingomyelin and about 3 wt. % negatively charged phospholipids, the molar ratio of lipid fraction to ApoA-I apolipoprotein is about 2:1 to 200:1, and the complex is a small or large discoid particle containing 2 to 4 ApoA-I equivalents.

[0051] In another specific embodiment, a lipoprotein complex that can be used in the methods of the present disclosure comprises an ApoA-I apolipoprotein and a lipid fraction, the lipid fraction consisting essentially of sphingomyelin and about 3 wt. % negatively charged phospholipids, the molar ratio of lipid fraction to ApoA-I apolipoprotein is about 2:1 to 200:1, and the complex is a small or large discoid particle containing 2 to 4 ApoA-I equivalents.

[0052] The complex based on HDL or HDL mimics can comprise a single type of lipid-binding protein or a mixture of two or more different lipid-binding proteins, which can be derived from the same or different species.Although not necessary, the complex preferably comprises lipid-binding proteins derived from the animal species being treated or corresponding to the amino acid sequence thereof, in order to avoid inducing immune response against the treatment.Therefore, lipid-binding proteins of human origin are preferably used for the treatment of human patients.The use of peptidomimetic apolipoproteins can also reduce or avoid immune response.

[0053] In some embodiments, the lipid component comprises two types of phospholipids: sphingomyelin (SM) and negatively charged phospholipids. Exemplary SMs and negatively charged lipids are described in Section 5.1.4.1.

[0054] The lipid component, including SM, can optionally contain small amounts of additional lipids. Virtually any type of lipid may be used, including, but not limited to, lysophospholipids, galactocerebrosides, gangliosides, cerebrosides, glycerides, triglycerides, and cholesterol and its derivatives.

[0055] If included, such optional lipids typically comprise less than about 15 wt% of the lipid fraction, but in some instances may comprise more optional lipids. In some embodiments, optional lipids comprise less than about 10 wt%, less than about 5 wt%, or less than about 2 wt%. In some embodiments, the lipid fraction does not comprise optional lipids.

[0056] In a specific embodiment, the phospholipid fraction contains egg SM or palmitoyl SM or phytosphingomyelin and DPPG in a weight ratio (SM:negatively charged phospholipid) ranging from 90:10 to 99:1, more preferably ranging from 95:5 to 98:2. In one embodiment, the weight ratio is 97:3.

[0057] The molar ratio of lipid to protein components of the complexes of the present disclosure may vary and will depend, among other factors, on the identity of the apolipoprotein that comprises the protein component, the identity and amount of lipid that comprises the lipid component, and the desired size of the complex. Because the biological activity of apolipoproteins such as ApoA-I is believed to be mediated by the amphipathic helices that comprise the apolipoprotein, it is convenient to use the equivalent of ApoA-I protein to represent the apolipoprotein fraction of the lipid:apolipoprotein molar ratio. It is generally accepted that ApoA-I contains 6-10 amphipathic helices depending on the method used to calculate the helices. Other apolipoproteins can be expressed in terms of the ApoA-I equivalent based on the number of amphipathic helices they contain. For example, ApoA-I, which typically exists as a disulfide-bridged dimer, is a soluble form of apolipoprotein that is soluble in water. M ApoA-I M can be expressed as 2 ApoA-I equivalents since each molecule contains twice as many amphipathic helices as ApoA-I molecules. Conversely, a peptide apolipoprotein containing a single amphipathic helix can be expressed as 1 / 10 to 1 / 6 ApoA-I equivalents since each molecule contains 1 / 10 to 1 / 6 the number of amphipathic helices as ApoA-I molecules. Generally, the molar ratio of lipid:ApoA-I equivalents (defined herein as "Ri") of the lipoprotein complex ranges from about 105:1 to 110:1. In some embodiments, Ri is about 108:1. Weight ratios can be obtained using phospholipids with a MW of approximately 650 to 800.

[0058] In some embodiments, the molar ratio of lipid:ApoA-I equivalents ("RSM") ranges from about 80:1 to about 110:1, such as from about 80:1 to about 100:1. In one specific example, the RSM of the complex can be about 82:1.

[0059] In some embodiments, the lipoprotein complex used in the methods of the present disclosure is a negatively charged complex comprising a protein fraction, preferably mature full-length ApoA-I, and a lipid fraction comprising neutral phospholipids, sphingomyelin (SM), and negatively charged phospholipids.

[0060] In a specific embodiment, the lipid component contains SM (e.g., egg SM, palmitoyl SM, phyto SM, or a combination thereof) and negatively charged phospholipid (e.g., DPPG) in a weight ratio (SM:negatively charged phospholipid) in the range of 90:10 to 99:1, more preferably in the range of 95:5 to 98:2, for example 97:3.

[0061] In specific embodiments, the ratio of protein to lipid components can range from about 1:2.7 to about 1:3, with 1:2.7 being preferred. This corresponds to a molar ratio of ApoA-I protein to lipid ranging from approximately 1:90 to 1:140. In some embodiments, the molar ratio of protein to lipid in the complex is from about 1:90 to about 1:120, from about 1:100 to about 1:140, or from about 1:95 to about 1:125.

[0062] In certain embodiments, the complex comprises CER-001, CSL-111, CSL-112, CER-522, or ETC-216. In a preferred embodiment, the complex is CER-001.

[0063] CER-001 as used in the literature and in the following examples refers to the complex described in Example 4 of WO2012 / 109162. WO2012 / 109162 refers to CER-001 as a complex having a lipoprotein weight:total phospholipid weight ratio of 1:2.7 and a SM:DPPG weight:weight ratio of 97:3. Example 4 of WO2012 / 109162 also describes its preparation method.

[0064] As used in the context of the disclosed methods and / or CER-001 dosing regimens, CER-001 refers to a lipoprotein complex whose individual components can vary by 20% from CER-001 as described in Example 4 of WO 2012 / 109162. In certain embodiments, the components of the lipoprotein complex vary by 10% from CER-001 as described in Example 4 of WO 2012 / 109162. Preferably, the components of the lipoprotein complex are as described in Example 4 of WO 2012 / 109162 (± allowed manufacturing tolerance variations). The SM in CER-001 can be natural or synthetic. In some embodiments, the SM is a natural SM, such as a natural SM as described in WO 2012 / 109162, such as chicken egg SM. In some embodiments, the SM is a synthetic SM, such as a synthetic SM described in WO 2012 / 109162, e.g., a synthetic palmitoyl sphingomyelin, e.g., as described in WO 2012 / 109162. Methods for synthesizing palmitoyl sphingomyelin are known in the art, e.g., as described in WO 2014 / 140787. The lipoprotein in CER-001, apolipoprotein AI (ApoA-I), preferably has an amino acid sequence corresponding to amino acids 25-267 of SEQ ID NO: 1 of WO 2012 / 109162 (identical to SEQ ID NO: 2 of the present application). ApoA-I can be purified from an animal source (particularly from a human source) or recombinantly produced. In a preferred embodiment, the ApoA-I in CER-001 is recombinant ApoA-I. CER-001 for use in the dosing regimens of the present disclosure is preferably highly homogenous, e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% homogenous, as reflected by a single peak in gel permeation chromatography. See, e.g., Section 6.4 of WO 2012 / 109162.

[0065] CSL-111 is a reconstituted human ApoA-I purified from plasma complexed with soybean phosphatidylcholine (SBPC) (Tardif et al., 2007, JAMA 297:1675-1682).

[0066] CSL-112 is a formulation of ApoA-I purified from plasma and reconstituted to form HDL suitable for intravenous infusion (Diditchenko et al., 2013, DOI 10.1161 / ATVBAHA.113.301981).

[0067] ETC-216 (also known as MDCO-216) is a recombinant ApoA-I ミラノ It is a lipid-depleted form of HDL that contains. See Nicholls et al., 2011, Expert Opin Biol Ther. 11(3):387-94. doi: 10.1517 / 14712598.2011.557061.

[0068] In another embodiment, a complex that can be used in the methods of the present disclosure is CER-522. CER-522 is a lipoprotein complex that contains a combination of three phospholipids and a 22 amino acid peptide, CT80522.

[0069] [ka]

[0070] The phospholipid component of CER-522 consists of egg sphingomyelin, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (dipalmitoylphosphatidylcholine, DPPC) and 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (dipalmitoylphosphatidyl-glycerol, DPPG) in a weight ratio of 48.5:48.5:3. The ratio of peptide to total phospholipid in the CER-522 complex is 1:2.5 (w / w).

[0071] In some embodiments, the lipoprotein complex is delipidated HDL. Most HDL in plasma is cholesterol-rich. Lipids in HDL can be depleted, e.g., partially and / or selectively depleted, e.g., to reduce its cholesterol content. In some embodiments, delipidated HDL can resemble small alpha, pre-beta-1, and other pre-beta forms of HDL. The process of selective depletion of HDL is described in Sacks et al., 2009, J Lipid Res. 50(5): 894-907.

[0072] In certain embodiments, the lipoprotein complexes comprise bioactive agent delivery particles as described in US Patent Application Publication No. 2004 / 0229794.

[0073] The bioactive agent delivery particle can include a lipid-binding polypeptide (e.g., an apolipoprotein as previously described in this section or in Section 5.1.2), a lipid bilayer (e.g., including one or more phospholipids as previously described in this section or in Section 5.1.4.1), and a bioactive agent (e.g., an anti-cancer agent), wherein the interior of the lipid bilayer includes a hydrophobic region, and the bioactive agent is associated with the hydrophobic region of the lipid bilayer. In some embodiments, the bioactive agent delivery particle is described in U.S. Patent Application Publication No. 2004 / 0229794.

[0074] In some embodiments, the bioactive agent delivery particles do not include a hydrophilic core.

[0075] In some embodiments, the bioactive agent delivery particles are disc-shaped (eg, having a diameter of about 7 to about 29 nm).

[0076] The bioactive agent delivery particle comprises a bilayer-forming lipid, such as a phospholipid (e.g., as previously described in this section or in Section 5.1.4.1). In some embodiments, the bioactive agent delivery particle comprises both bilayer-forming and non-bilayer-forming lipids. In some embodiments, the lipid bilayer of the bioactive agent delivery particle comprises a phospholipid. In one embodiment, the phospholipids incorporated within the delivery particle include dimyristoyl phosphatidylcholine (DMPC) and dimyristoyl phosphatidylglycerol (DMPG). In one embodiment, the lipid bilayer comprises DMPC and DMPG in a molar ratio of 7:3.

[0077] In some embodiments, the lipid-binding polypeptide is an apolipoprotein (e.g., as described in this section or in section 5.1.2). The predominant interaction between the lipid-binding polypeptide, e.g., apolipoprotein molecule, and the lipid bilayer is generally a hydrophobic interaction between residues on the hydrophobic face of the amphiphilic structure, e.g., the α-helix of the lipid-binding polypeptide, and the fatty acyl chain of the lipid on the outer surface of the particle periphery. The bioactive agent delivery particle may include exchangeable and / or non-exchangeable apolipoproteins. In one embodiment, the lipid-binding polypeptide is ApoA-I.

[0078] In some embodiments, the bioactive agent delivery particles include lipid-binding polypeptide molecules, such as apolipoprotein molecules, that have been modified to increase the stability of the particles. In one embodiment, the modifications include the introduction of cysteine ​​residues to form intra- and / or intermolecular disulfide bonds.

[0079] In another embodiment, the bioactive agent delivery particles include chimeric lipid-binding polypeptide molecules, e.g., chimeric apolipoprotein molecules, having one or more attached functional moieties that can enhance or act synergistically with the activity of the bioactive agent incorporated within the delivery particle, e.g., one or more targeting moieties and / or one or more moieties having a desired biological activity, such as antimicrobial activity.

[0080] 5.1.2.ApoA-I formulation In one aspect, the disclosure relates to ApoA-I formulations comprising ApoA-I and one or more lipids, wherein the ApoA-I and lipids are in the form of a lipoprotein complex.

[0081] The ApoA-I can be any such apolipoprotein described in Section 5.1.3.1, including, inter alia, ApoA-I having the amino acid sequence of amino acids 25 to 267 of SEQ ID NO:2 and / or recombinantly expressed ApoA-I.

[0082] The lipids may be any one or more of those described in Section 5.1.4.1. The lipids may include neutral lipids and / or negatively charged lipids. The neutral lipids may include or consist of sphingomyelin, such as natural sphingomyelin (e.g., chicken egg sphingomyelin) and / or synthetic sphingomyelin (e.g., palmitoyl sphingomyelin). The negatively charged lipids may include or consist of 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] ("DPPG") or a salt thereof.

[0083] The lipids can include neutral phospholipids and negatively charged phospholipids in any weight or molar ratio described herein. The lipids may be comprised of 95-99% by weight neutral phospholipids and 1-5% by weight negatively charged phospholipids, for example, 96-98% by weight neutral phospholipids and 2-4% by weight negatively charged phospholipids, or 97% by weight neutral phospholipids and 3% by weight negatively charged phospholipids.

[0084] The formulation can include ApoA-I and lipid in any weight or molar ratio described herein. For example, the molar ratio of the negatively charged lipid component to neutral lipid to ApoA-I in the formulation is 2-6:90-120:1. Exemplary formulations can include an ApoA-I to lipid ratio in the range of 1:2-1:3 by weight, for example about 1:2.7 by weight.

[0085] The formulation can be used in methods of treating subjects with one or more symptoms associated with leukocytosis, subjects with endothelial dysfunction, subjects with or at risk of developing carditis, subjects with or at risk of developing leukocytosis, subjects experiencing an acute coronary syndrome or stroke, and / or subjects who have experienced an acute coronary syndrome or stroke, such as the methods described in Sections 5.2-5.4. The formulation can also be used for use in the treatment of such diseases or disorders. The formulation can also be for use in the manufacture of a medicament for treating such diseases or disorders.

[0086] Examples of ApoA-I formulations and uses thereof include those described as numbered embodiments 1-72 of Group 2.

[0087] 5.1.3. Lipid-binding protein molecules The lipid-binding protein molecules that can be used in the complexes described herein include apolipoproteins, such as those described in Section 5.1.3.1, and apolipoprotein mimetic peptides, such as those described in Section 5.1.3.2. In some embodiments, the complexes comprise a mixture of lipid-binding protein molecules. In some embodiments, the complexes comprise a mixture of one or more lipid-binding protein molecules and one or more apolipoprotein mimetic peptides.

[0088] In some embodiments, the complex comprises 1-8 ApoA-I equivalents (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 1-7, 1-6, 1-5, 1-4, 1-3, 1-2, 2-8, 2-6, 2-4, 4-6, or 4-8 ApoA-I equivalents). Lipid binding proteins can be expressed in terms of ApoA-I equivalents based on the number of amphipathic helices they contain. For example, ApoA-I, which typically exists as a disulfide-bridged dimer, is a lipid-binding protein that is capable of binding to at least one of the 20 ApoA-I equivalents. M ApoA-I M can be expressed as 2 ApoA-I equivalents, since each molecule contains twice the number of amphipathic helices as an ApoA-I molecule. Conversely, peptidomimetics containing a single amphipathic helix can be expressed as 1 / 10-1 / 6 ApoA-I equivalents, since each molecule contains 1 / 10-1 / 6 the number of amphipathic helices as an ApoA-I molecule.

[0089] Apolipoproteins Suitable apolipoproteins that can be included in the lipid-binding protein-based complex include apolipoproteins ApoA-I, ApoA-II, ApoA-IV, ApoA-V, ApoB, ApoC-I, ApoC-II, ApoC-III, ApoD, ApoE, ApoJ, ApoH, and any combination of two or more of the above. Polymorphic forms, isoforms, variants, and mutants, as well as truncated forms of the above apolipoproteins, the most common of which are apolipoprotein AI Milano (ApoA-IM), apolipoprotein AI Paris (ApoA-IP), and apolipoprotein AI Zaragoza (ApoA-IZ), are also known and can also be used (see, for example, US Patent Publication 2003 / 0181372). Apolipoproteins can be in the form of monomers or dimers, which can be homodimers or heterodimers. For example, ApoA-I (Duverger et al., 1996, Arterioscler. Thromb. Vasc. Biol. 16(12):1424-29), ApoA-IM (Franceschini et al., 1985, J. Biol. Chem. 260:1632-35), ApoA-IP (Daum et al., 1999, J. Mol. Med. 77:614-22), ApoA-II (Shelness et al., 1985, J. Biol. Chem. 260(14):8637-46, Shelness et al., 1984, J. Biol. Chem. 259(15):9929-35), ApoA-IV (Duverger et al., 1991, Euro. J. Biochem. 201(2):373-83), ApoE (McLean et al., 1983, J. Biol. Chem. 258(14):8993-9000), ApoJ, and homo- and heterodimers (where possible) of ApoH may be used.

[0090] Apolipoproteins can be modified in their primary sequence to make them less susceptible to oxidation, for example, as described in US Patent Publications 2008 / 0234192 and 2013 / 0137628, and US Patents 8,143,224 and 8,541,236. Apolipoproteins can contain residues that correspond to elements that facilitate their isolation, such as His tags, or other elements designed for other purposes. Preferably, the apolipoproteins in the complex are soluble in biological fluids (e.g., lymph, cerebrospinal fluid, vitreous fluid, aqueous humor, blood, or blood fractions (e.g., serum or plasma).

[0091] In some embodiments, the complex comprises a covalently linked lipid-binding protein monomer, e.g., dimeric apolipoprotein AI Milano, a variant of ApoA-I that contains cysteines. The cysteines allow for the formation of disulfide bridges, which can result in the formation of homodimers or heterodimers (e.g., ApoA-I Milano-ApoA-II).

[0092] In some embodiments, the apolipoprotein molecule includes an ApoA-I, ApoA-II, ApoA-IV, ApoA-V, ApoB, ApoC-I, ApoC-II, ApoC-III, ApoD, ApoE, ApoJ, or ApoH molecule, or a combination thereof.

[0093] In some embodiments, the apolipoprotein molecule comprises or consists of an ApoA-I molecule. In some embodiments, the ApoA-I molecule is a human ApoA-I molecule. In some embodiments, the ApoA-I molecule is recombinant. In some embodiments, the ApoA-I molecule is not ApoA-I Milano.

[0094] In some embodiments, the ApoA-I molecule is an Apolipoprotein AI Milano (ApoA-IM), Apolipoprotein AI Paris (ApoA-IP), or Apolipoprotein AI Zaragoza (ApoA-IZ) molecule.

[0095] Apolipoproteins can be purified from animal sources (especially human sources) or recombinantly produced, as is well known in the art.See, for example, Chung et al., 1980, J. Lipid Res. 21(3):284-91; Cheung et al., 1987, J. Lipid Res. 28(8):913-29.See also U.S. Patent Nos. 5,059,528, 5,128,318, 6,617,134, U.S. Patent Publication Nos. 2002 / 0156007, 2004 / 0067873, 2004 / 0077541, and 2004 / 0266660, and WO 2008 / 104890 and WO 2007 / 023476. Other purification methods are also possible, for example as described in WO 2012 / 109162, the disclosure of which is incorporated herein by reference in its entirety.

[0096] The apolipoprotein may be in a prepro form, a pro form, or a mature form. For example, the complex may include ApoA-I (e.g., human ApoA-I), in which ApoA-I is preproApoA-I, proApoA-I, or mature ApoA-I. In some embodiments, the complex includes ApoA-I having at least 90% sequence identity to SEQ ID NO:1.

[0097] [ka]

[0098] In other embodiments, the complex comprises an ApoA-I having at least 95% sequence identity to SEQ ID NO: 1. In other embodiments, the complex comprises an ApoA-I having at least 98% sequence identity to SEQ ID NO: 1. In other embodiments, the complex comprises an ApoA-I having at least 99% sequence identity to SEQ ID NO: 1. In other embodiments, the complex comprises an ApoA-I having 100% sequence identity to SEQ ID NO: 1.

[0099] In some embodiments, the complex comprises 1-8 apolipoprotein molecules (e.g., 1-6, 1-4, 1-2, 2-8, 2-6, 2-4, 4-8, 4-6, or 6-8 apolipoprotein molecules). In some embodiments, the complex comprises 1 apolipoprotein molecule. In some embodiments, the complex comprises 2 apolipoprotein molecules. In some embodiments, the complex comprises 3 apolipoprotein molecules. In some embodiments, the complex comprises 4 apolipoprotein molecules. In some embodiments, the complex comprises 5 apolipoprotein molecules. In some embodiments, the complex comprises 6 apolipoprotein molecules. In some embodiments, the complex comprises 7 apolipoprotein molecules. In some embodiments, the complex comprises 8 apolipoprotein molecules.

[0100] The apolipoprotein molecule can include a chimeric apolipoprotein that includes an apolipoprotein and one or more attached functional moieties, such as one or more CRN-001 complexes, one or more targeting moieties, a moiety with a desired biological activity, an affinity tag to aid in purification, and / or a reporter molecule for characterization or localization studies. The attached biologically active moiety can have an activity that can enhance and / or synergize with the biological activity of the compound incorporated into the complex of the present disclosure. For example, the biologically active moiety can have an antimicrobial (e.g., antifungal, antibacterial, antiprotozoal, bacteriostatic, fungistatic, or antiviral) activity. In one embodiment, the attached functional moiety of the chimeric apolipoprotein is not in contact with a hydrophobic surface of the complex. In another embodiment, the attached functional moiety is in contact with a hydrophobic surface of the complex. In some embodiments, the functional moiety of the chimeric apolipoprotein can be endogenous to the natural protein. In some embodiments, the chimeric apolipoprotein includes a ligand or sequence that can be recognized by or interact with a cell surface receptor or other cell surface moiety.

[0101] In one embodiment, the chimeric apolipoprotein includes a targeting moiety that is not endogenous to the native apolipoprotein, such as, for example, S. cerevisiae α-mating factor peptide, folic acid, transferrin, or lactoferrin. In another embodiment, the chimeric apolipoprotein includes a moiety that has a desired biological activity that enhances and / or synergizes with the activity of a compound incorporated into the complex of the present disclosure. In one embodiment, the chimeric apolipoprotein can include a functional moiety that is endogenous to the apolipoprotein. One example of an apolipoprotein endogenous functional moiety is the endogenous targeting moiety formed approximately by amino acids 130-150 of human ApoE, which includes the receptor binding region recognized by members of the low density lipoprotein receptor family. Other examples of apolipoprotein endogenous functional moieties include the region of ApoB-100 that interacts with the low density lipoprotein receptor and the region of ApoA-I that interacts with scavenger receptor B1. In other embodiments, functional moieties can be added synthetically or recombinantly to produce chimeric apolipoproteins. Another example is an apolipoprotein having a prepro or pro sequence from another preproapolipoprotein (e.g., a prepro sequence from proproapoA-II replaces the prepro sequence of preproapoAI). Another example is an apolipoprotein in which some of the amphipathic sequence segments are replaced by other amphipathic sequence segments from another apolipoprotein.

[0102] As used herein, "chimera" refers to two or more molecules that can exist separately and are linked together to form a single molecule that has all the desired functionality of its constituent molecules. The constituent molecules of a chimeric molecule can be synthetically combined by chemical conjugation, or, if the constituent molecules are all polypeptides or analogs thereof, the polynucleotides encoding the polypeptides can be recombinantly fused together so that a single continuous polypeptide is expressed. Such chimeric molecules are called fusion proteins. A "fusion protein" is a chimeric molecule in which the constituent molecules are all polypeptides and are attached (fused) to each other so that the chimeric molecule forms a continuous single chain. The various components can be directly attached to each other or can be coupled through one or more linkers. One or more segments of the various components can be inserted, for example, into the sequence of the apolipoprotein, or, as another example, can be added to the N-terminus or C-terminus of the apolipoprotein sequence. For example, the fusion protein can include an antibody light chain, an antibody fragment, a heavy chain antibody, or a single domain antibody.

[0103] In some embodiments, the chimeric apolipoprotein is prepared by chemically conjugating the apolipoprotein with the functional moiety to be attached. Means for chemically conjugating molecules are well known to those skilled in the art. Such means vary depending on the structure of the moiety to be attached, but would be readily ascertainable to those skilled in the art. Polypeptides typically contain a variety of functional groups, such as carboxylic acid (-COOH), free amino (-NH2), or sulfhydryl (-SH) groups, that can be reacted with appropriate functional groups on the functional moiety or on the linker to attach the moiety thereto. The functional moiety may be attached to a functional group on the N-terminus, C-terminus, or an internal residue (i.e., a residue intermediate the N-terminus and C-terminus) of the apolipoprotein molecule. Alternatively, the apolipoprotein and / or the tagging moiety may be derivatized to expose or attach additional reactive functional groups.

[0104] In some embodiments, a fusion protein comprising a polypeptide functional moiety is synthesized using a recombinant expression system. Typically, this involves creating a nucleic acid (e.g., DNA) sequence that encodes an apolipoprotein and a functional moiety such that the two polypeptides are in frame when expressed, placing the DNA under the control of a promoter, expressing the protein in a host cell, and isolating the expressed protein.

[0105] The nucleic acid encoding the chimeric apolipoprotein can be incorporated into a recombinant expression vector in a form suitable for expression in a host cell. As used herein, an "expression vector" is a nucleic acid that can be transcribed and translated into a polypeptide when introduced into a suitable host cell. The vector can include regulatory sequences such as promoters, enhancers, or other expression control elements (e.g., polyadenylation signals). Such regulatory sequences are known to those skilled in the art (see, for example, Goeddel, 1990, Gene Expression Technology: Meth. Enzymol. 185, Academic Press, San Diego, Calif.; Berger and Kimmel, Guide to Molecular Cloning Techniques, Methods in Enzymology 152 Academic Press, Inc., San Diego, Calif.; Sambrook et al., 1989, Molecular Cloning--A Laboratory Manual (2nd ed.) Vol. 1-3, Cold Spring Harbor Laboratory, Cold Spring Harbor Press, NY, etc.).

[0106] In some embodiments, the apolipoprotein is modified such that the modification increases the stability of the complex and confers or increases the targeting ability when the apolipoprotein is incorporated into the complex of the present disclosure. In one embodiment, the modification includes the introduction of a cysteine ​​residue into the apolipoprotein molecule, for example by site-directed mutagenesis, to allow the formation of intramolecular or intermolecular disulfide bonds. In another embodiment, a chemical crosslinker is used to form an intermolecular linkage between apolipoprotein molecules to enhance the stability of the complex. The intermolecular crosslink prevents or reduces the dissociation of the apolipoprotein molecule from the complex and / or prevents the displacement of endogenous apolipoprotein molecules in the individual to which the complex is administered. In other embodiments, the apolipoprotein is modified by either chemical derivatization or site-directed mutagenesis of one or more amino acid residues to confer targeting ability or recognition by cell surface receptors.

[0107] The complex can be targeted to a specific cell surface receptor by engineering receptor recognition characteristics into the apolipoprotein. For example, the complex can be targeted to a particular cell type known to carry a particular type of infectious agent, for example, by modifying the apolipoprotein to allow it to interact with a receptor on the surface of the targeted cell type. For example, the complex can be targeted to macrophages by altering the apolipoprotein to confer recognition by macrophage endocytic class A scavenger receptor (SR-A). SR-A binding ability can be conferred to the complex by modifying the apolipoprotein by replacing one or more positively charged amino acids with neutral or negatively charged amino acids by site-directed mutagenesis. SR-A recognition can also be conferred by preparing chimeric apolipoproteins that contain N- or C-terminal extensions with ligands recognized by SR-A or amino acid sequences with a high concentration of negatively charged residues. Complexes containing apolipoproteins can also interact with apolipoprotein receptors, such as, but not limited to, the ABCA1 receptor, the ABCG1 receptor, Megalin, Cubulin, and the HDL receptor, e.g., SR-B1.

[0108] 5.1.3.2. Apolipoprotein Mimetics Peptides, peptide analogs, and agonists that mimic the activity of apolipoproteins (collectively referred to herein as "apolipoprotein peptidomimetics") can also be used in the complexes described herein, either alone or in combination with one or more other lipid-binding proteins. Peptides and peptide analogs corresponding to apolipoproteins, as well as ApoA-I, ApoA-I, and ApoA-I are suitable for inclusion in the complexes and compositions described herein. MNon-limiting examples of agonists that mimic the activity of ApoA-II, ApoA-IV, and ApoE are described in U.S. Pat. Nos. 6,004,925, 6,037,323, and 6,046,166 (issued to Dasseux et al.), U.S. Pat. No. 5,840,688 (issued to Tso), U.S. Pat. No. 6,743,778 (issued to Kohno), U.S. Patent Publication No. 2004 / 0266671, U.S. Pat. No. 6,743,778 (issued to Kohno), the disclosures of which are incorporated by reference in their entireties herein. US Patent Publication Nos. 2004 / 0254120, 2003 / 0171277, and 2003 / 0045460 (Fogelman), US Patent Publication No. 2006 / 0069030 (Bachovchin), US Patent Publication No. 2003 / 0087819 (Bielicki), US Patent Publication No. 2009 / 0081293 (Murase et al.), and WO 2010 / 093918 (Dasseux et al.). These peptides and peptide analogs may be composed of L- or D-amino acids or mixtures of L- and D-amino acids. They may also contain one or more non-peptide or amide bonds, such as one or more of the well-known peptide / amide isosteres. Such apolipoprotein peptidomimetics can be synthesized or produced using any of the techniques for peptide synthesis known in the art, including, for example, the techniques described in U.S. Pat. Nos. 6,004,925, 6,037,323, and 6,046,166.

[0109] In some embodiments, the lipid binding protein molecule comprises an apolipoprotein peptidomimetic molecule, and optionally one or more apolipoprotein molecules, such as those described above.

[0110] In some embodiments, the apolipoprotein peptidomimetic molecule comprises an ApoA-I peptidomimetic, an ApoA-II peptidomimetic, an ApoA-IV peptidomimetic, or an ApoE peptidomimetic, or a combination thereof.

[0111] 5.1.4. Amphiphilic molecules Amphipathic molecules are molecules that possess both hydrophobic (non-polar) and hydrophilic (polar) elements. Amphipathic molecules that can be used in the complexes described herein include lipids (e.g., as described in Section 5.1.4.1), detergents (e.g., as described in Section 5.1.4.2), fatty acids (e.g., as described in Section 5.1.4.3), and non-polar molecules covalently attached to polar molecules, such as, but not limited to, sugars or nucleic acids, and sterols (e.g., as described in Section 5.1.4.4).

[0112] A complex can include a single class of amphipathic molecule (e.g., a single species of phospholipid or a mixture of phospholipids) or can contain a combination of classes of amphipathic molecules (e.g., phospholipids and detergents). A complex can contain one amphipathic molecule or a combination of amphipathic molecules configured to facilitate solubilization of lipid-binding protein molecules.

[0113] In some embodiments, the amphipathic molecules included include phospholipids, detergents, fatty acids, nonpolar moieties covalently attached to sugars or sterols, or combinations thereof (e.g., selected from the types of amphipathic molecules described above).

[0114] In some embodiments, the amphipathic molecules comprise or consist of phospholipid molecules. In some embodiments, the phospholipid molecules comprise negatively charged phospholipids, neutral phospholipids, positively charged phospholipids, or a combination thereof. In some embodiments, the phospholipid molecules contribute 1-3 net charges per apolipoprotein molecule in the complex. In some embodiments, the net charge is a negative net charge. In some embodiments, the net charge is a positive net charge. In some embodiments, the phospholipid molecules consist of a combination of negatively charged and neutral phospholipids. In some embodiments, the molar ratio of negatively charged phospholipids to neutral phospholipids ranges from 1:1 to 1:3. In some embodiments, the molar ratio of negatively charged phospholipids to neutral phospholipids is about 1:1 or about 1:2.

[0115] In some embodiments, the amphipathic molecule comprises neutral phospholipids and negatively charged phospholipids in a weight ratio of 95:5 to 99:1.

[0116] Lipids A lipid-binding protein-based complex can include one or more lipids. In various embodiments, the one or more lipids can be saturated and / or unsaturated, natural and / or synthetic, charged or uncharged, zwitterionic or not. In some embodiments, the lipid molecules (e.g., phospholipid molecules) can together contribute a net charge of 1-3 (e.g., 1-3, 1-2, 2-3, 1, 2, or 3) per lipid-binding protein molecule in the complex. In some embodiments, the net charge is negative. In other embodiments, the net charge is positive.

[0117] In some embodiments, the lipid comprises a phospholipid. The phospholipid can have two acyl chains that are the same or different (e.g., chains with different numbers of carbon atoms, different degrees of saturation between the acyl chains, different branching of the acyl chains, or combinations thereof). The lipid can also be modified to contain a fluorescent probe (e.g., as described at avantilipids.com / product-category / products / fluorescent-lipids / ). Preferably, the lipid comprises at least one phospholipid.

[0118] Phospholipids can have unsaturated or saturated acyl chains ranging from about 6 to about 24 carbon atoms (e.g., 6-20, 6-16, 6-12, 12-24, 12-20, 12-16, 16-24, 16-20, or 20-24). In some embodiments, phospholipids for use in the conjugates of the present disclosure have one or two acyl chains of 12, 14, 16, 18, 20, 22, or 24 carbons (e.g., two acyl chains of the same length or two acyl chains of different lengths).

[0119] Non-limiting examples of acyl chains present in commonly occurring fatty acids that may be included in phospholipids are provided in Table 1 below.

[0120] [Table 1]

[0121] Lipids that may be present in the complexes of the present disclosure include, but are not limited to, small alkyl chain phospholipids, egg phosphatidylcholine, soy phosphatidylcholine, dipalmitoyl phosphatidylcholine, dimyristoyl phosphatidylcholine, distearoyl phosphatidylcholine, 1-myristoyl-2-palmitoyl phosphatidylcholine, 1-palmitoyl-2-myristoyl phosphatidylcholine, 1-palmitoyl-2-stearoyl phosphatidylcholine, 1-stearoyl-2-palmitoyl phosphatidylcholine, dioleoyl phosphatidylcholine, dioleoyl phosphatidylethanolamine, dilauroyl phosphatidylglycerol. Phosphatidylcholine, phosphatidylserine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylglycerol, diphosphatidylglycerol, for example, dimyristoylphosphatidylglycerol, dipalmitoylphosphatidylglycerol, distearoylphosphatidylglycerol, dioleoylphosphatidylglycerol, dimyristoylphosphatidic acid, dipalmitoylphosphatidic acid, dimyristoylphosphatidylethanolamine, dipalmitoylphosphatidylethanolamine, dimyristoylphosphatidylserine, dipalmitoylphosphatidylserine Examples of lipids that can be used include phosphatidylserine, brain sphingomyelin, palmitoyl sphingomyelin, dipalmitoyl sphingomyelin, egg sphingomyelin, milk sphingomyelin, phytosphingomyelin, distearoyl sphingomyelin, dipalmitoyl phosphatidylglycerol salts, phosphatidic acid, galactocerebroside, ganglioside, cerebroside, dilauryl phosphatidylcholine, (1,3)-D-mannosyl-(1,3) diglyceride, aminophenylglycosides, 3-cholesteryl-6'-(glycosylthio)hexyl ether glycolipids, and cholesterol and its derivatives. Synthetic lipids such as synthetic palmitoyl sphingomyelin or N-palmitoyl-4-hydroxysphinganine-1-phosphocholine (a form of phytosphingomyelin) can be used to minimize lipid oxidation.

[0122] In some embodiments, the lipid-binding protein-based complex comprises two types of phospholipids, namely, a neutral lipid, such as lecithin and / or sphingomyelin (abbreviated as SM), and a charged phospholipid (e.g., a negatively charged phospholipid). A "neutral" phospholipid has a net charge of about zero at physiological pH. In many embodiments, the neutral phospholipid is zwitterionic, although other types of net neutral phospholipids are known and can be used. In some embodiments, the molar ratio of charged phospholipid (e.g., negatively charged phospholipid) to neutral phospholipid is in the range of 1:1 to 1:3, e.g., about 1:1, about 1:2, or about 1:3.

[0123] The neutral phospholipids can include, for example, one or both of lecithin and / or SM, and can optionally include other neutral phospholipids. In some embodiments, the neutral phospholipids include lecithin but do not include SM. In other embodiments, the neutral phospholipids include SM but do not include lecithin. In still other embodiments, the neutral phospholipids include both lecithin and SM. All of these specific exemplary embodiments can include neutral phospholipids in addition to lecithin and / or SM, although many embodiments do not include such additional neutral phospholipids.

[0124] The expression "SM" as used herein includes sphingomyelin derived from or obtained from natural sources, as well as analogs and derivatives of naturally occurring SM that are not susceptible to hydrolysis by LCAT as naturally occurring SM is.SM is a phospholipid that is very similar in structure to lecithin, but unlike lecithin, it does not have a glycerol backbone and therefore does not have ester linkages that attach acyl chains.Instead, SM has a ceramide backbone, with amide bonds linking the acyl chains.SM can be obtained, for example, from milk, eggs, or brain.An analog or derivative of SM can also be used. Non-limiting examples of useful SM analogs and derivatives include, but are not limited to, palmitoyl sphingomyelin, N-palmitoyl-4-hydroxysphinganine-1-phosphocholine (a form of phytosphingomyelin), palmitoyl sphingomyelin, stearoyl sphingomyelin, D-erythro-N-16:0-sphingomyelin and its dihydro isomer, D-erythro-N-16:0-dihydro-sphingomyelin. To generate a more homogeneous complex with fewer contaminants and / or oxidation products than sphingolipids of animal origin, synthetic SMs such as synthetic palmitoyl sphingomyelin or N-palmitoyl-4-hydroxysphinganine-1-phosphocholine (phytosphingomyelin) can be used. Methods for synthesizing SMs are described in U.S. Patent Publication No. 2016 / 0075634.

[0125] Sphingomyelin isolated from natural sources can be artificially enriched for one particular saturated or unsaturated acyl chain. For example, milk sphingomyelin (Avanti Phospholipid, Alabaster, Alabama) is characterized by long saturated acyl chains (i.e., acyl chains with 20 or more carbon atoms). In contrast, egg sphingomyelin is characterized by short saturated acyl chains (i.e., acyl chains with fewer than 20 carbon atoms). For example, only about 20% of milk sphingomyelin contains C16:0 (16 carbons, saturated) acyl chains, while about 80% of egg sphingomyelin contains C16:0 acyl chains. Solvent extraction can be used to enrich the composition of milk sphingomyelin to have an acyl chain composition comparable to egg sphingomyelin, or vice versa.

[0126] SM can be semi-synthetic to have a specific acyl chain. For example, milk sphingomyelin can be first purified from milk, and then one specific acyl chain, e.g., the C16:0 acyl chain, can be cleaved and replaced by another acyl chain. SM can also be fully synthetic, e.g., by large-scale synthesis. See, e.g., Dong et al., U.S. Pat. No. 5,220,043, entitled Synthesis of D-erythro-sphingomyelins, published June 15, 1993; Weis, 1999, Chem. Phys. Lipids 102 (1-2):3-12. SM can be fully synthetic, e.g., as described in U.S. Patent Publication No. 2014 / 0275590.

[0127] The length and saturation level of the acyl chains comprising the semi-synthetic or synthetic SM can be selectively varied. The acyl chains can be saturated or unsaturated and can contain from about 6 to about 24 carbon atoms. Each chain can contain the same number of carbon atoms, or each chain can contain a different number of carbon atoms. In some embodiments, the semi-synthetic or synthetic SM comprises mixed acyl chains, such that one chain is saturated and one chain is unsaturated. In such mixed acyl chain SM, the chain lengths can be the same or different. In other embodiments, the acyl chains of the semi-synthetic or synthetic SM are either both saturated or both unsaturated. Again, the chains can contain the same or different numbers of carbon atoms. In some embodiments, both acyl chains comprising the semi-synthetic or synthetic SM are identical. In specific embodiments, the chains correspond to the acyl chains of naturally occurring fatty acids, such as oleic acid, palmitic acid, or stearic acid. In another embodiment, SM with saturated or unsaturated functionalized chains is used. In another specific embodiment, both acyl chains are saturated and contain from 6 to 24 carbon atoms. Non-limiting examples of acyl chains present in commonly occurring fatty acids that can be included in semi-synthetic and synthetic SMs are provided in Table 1 above.

[0128] In some embodiments, the SM is palmitoyl SM, eg, synthetic palmitoyl SM having a C16:0 acyl chain, or egg SM that contains palmitoyl SM as a major component.

[0129] In a specific embodiment, a functionalized SM, such as phytosphingomyelin, is used.

[0130] Lecithin can be derived from or isolated from natural sources, or it can be obtained by synthesis.The examples of suitable lecithin isolated from natural sources include, but are not limited to, egg phosphatidylcholine and soybean phosphatidylcholine.Further non-limiting examples of suitable lecithin include dipalmitoyl phosphatidylcholine, dimyristoyl phosphatidylcholine, distearoyl phosphatidylcholine, 1-myristoyl-2-palmitoyl phosphatidylcholine, 1-palmitoyl-2-myristoyl phosphatidylcholine, 1-palmitoyl-2-stearoyl phosphatidylcholine, 1-stearoyl-2-palmitoyl phosphatidylcholine, 1-palmitoyl-2-oleoyl phosphatidylcholine, 1-oleoyl-2-palmityl phosphatidylcholine, dioleoyl phosphatidylcholine and its ether derivatives or analogues.

[0131] Lecithin derived from or isolated from natural sources can be enriched to contain designated acyl chains. In embodiments using semi-synthetic or synthetic lecithin, the identity of the acyl chain can be selectively varied as described above in relation to SM. In some embodiments of the complexes described herein, both acyl chains on the lecithin are the same. In some embodiments of the complexes containing both SM and lecithin, the acyl chains of the SM and lecithin are all the same. In specific embodiments, the acyl chains correspond to the acyl chains of myristic acid, palmitic acid, oleic acid, or stearic acid.

[0132] The complexes of the present disclosure can include one or more negatively charged phospholipids (e.g., alone or in combination with one or more neutral phospholipids). As used herein, a "negatively charged phospholipid" is a phospholipid that has a net negative charge at physiological pH. The negatively charged phospholipid can include a single type of negatively charged phospholipid or a mixture of two or more differently negatively charged phospholipids. In some embodiments, the charged phospholipid is a negatively charged glycerophospholipid. Specific examples of suitable negatively charged phospholipids include, but are not limited to, 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)], phosphatidylglycerol, phosphatidylinositol, phosphatidylserine, phosphatidic acid, and salts thereof (e.g., sodium or potassium salts). In some embodiments, the negatively charged phospholipid includes one or more of phosphatidylinositol, phosphatidylserine, phosphatidylglycerol, and / or phosphatidic acid. In a specific embodiment, the negatively charged phospholipid comprises or consists of a salt of phosphatidylglycerol or a salt of phosphatidylinositol. In another specific embodiment, the negatively charged phospholipid comprises or consists of 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] or DPPG, or a salt thereof.

[0133] Negatively charged phospholipids can be obtained from natural sources or prepared by chemical synthesis. In embodiments using synthetic negatively charged phospholipids, the identity of the acyl chain can be selectively varied as described above with respect to SM. In some embodiments of the complexes of the present disclosure, both acyl chains on the negatively charged phospholipid are the same. In some embodiments, the acyl chains of all types of phospholipids included in the complexes of the present disclosure are all the same. In a specific embodiment, the complexes include negatively charged phospholipids and / or SMs that all have C16:0 or C16:1 acyl chains. In a specific embodiment, the fatty acid moiety of the SM is predominantly C16:1 palmitoyl. In one specific embodiment, the acyl chains of the charged phospholipids, lecithin and / or SM correspond to the acyl chains of palmitic acid. In yet another specific embodiment, the acyl chains of the charged phospholipids, lecithin and / or SM correspond to the acyl chains of oleic acid.

[0134] Examples of positively charged phospholipids that can be included in the complexes of the present disclosure include N1-[2-((1S)-1-[(3-aminopropyl)amino]-4-[di(3-amino-propyl)amino]butylcarboxamido)ethyl]-3,4-di[oleyloxy]-benzamide, 1,2-di-O-octadecenyl-3-trimethylammonium propane, 1,2-dimyristoleoyl-sn-glycero-3-ethylphosphocholine, 1-palmitoyl-2-oleoyl-sn-glyceroyl ... 1,2-Dioleoyl-sn-glycero-3-ethylphosphocholine, 1,2-Dioleoyl-sn-glycero-3-ethylphosphocholine, 1,2-Distearoyl-sn-glycero-3-ethylphosphocholine, 1,2-Dipalmitoyl-sn-glycero-3-ethylphosphocholine, 1,2-Dimyristoyl-sn-glycero-3-ethylphosphocholine, 1,2-Dilauroyl-sn-glycero-3-ethylphosphocholine, 1,2-Dilauroyl-sn-glycero-3-ethylphosphocholine, 1,2-Dioleoyl-sn-glycero-3-ethylphosphocholine, 1,2-Dimyristoyl-3-dimethylammonium-propane, 1,2-Dipalmitoyl-3-dimethylammonium-propane, N-(4-carboxybenzyl)-N,N-dimethyl-2,3-bis(oleoyloxy)propan-1-aminium, 1,2-Dioleoyl-3-trimethylammonium-propane, 1,2-Dioleoyl-3-trimethylammonium-propane, 1,2-Stearoyl-3-trimethyl Ammonium-propane, 1,2-dipalmitoyl-3-trimethylammonium-propane, 1,2-dimyristoyl-3-trimethylammonium-propane, N-[1-(2,3-dimyristyloxy)propyl]-N,N-dimethyl-N-(2-hydroxyethyl)ammonium bromide, N,N,N-trimethyl-2-bis[(1-oxo-9-octadecenyl)oxy]-(Z,Z)-1propaneaminium methyl sulfate, and salts thereof (e.g., chloride or bromide salts).

[0135] The lipids used are preferably at least 95% pure and / or have reduced levels of oxidizing agents, including but not limited to peroxides. Lipids obtained from natural sources preferably have less polyunsaturated fatty acid moieties and / or fatty acid moieties that are less susceptible to oxidation. The level of oxidation in a sample can be determined using iodometric methods that provide peroxide values, which are expressed as milliequivalents of isolated iodine per kg of sample, abbreviated as meq O / kg. See, for example, Gray, 1978, Measurement of Lipid Oxidation: A Review, Journal of the American Oil Chemists Society 55:539-545; Heaton, FW and Ur, Improved Iodometric Methods for the Determination of Lipid Peroxides, 1958, Journal of the Science of Food and Agriculture 9:781-786. Preferably, the level of oxidation, or peroxide levels, is low, for example, less than 5 meq O / kg, less than 4 meq O / kg, less than 3 meq O / kg, or less than 2 meq O / kg.

[0136] In some embodiments, the complex may contain a small amount of additional lipid. Virtually any type of lipid may be used, including but not limited to lysophospholipids, galactocerebrosides, gangliosides, cerebrosides, glycerides, triglycerides, and sterols and sterol derivatives (e.g., plant sterols, animal sterols such as cholesterol, or sterol derivatives such as cholesterol derivatives). For example, the complex of the present disclosure may contain cholesterol or cholesterol derivatives, such as cholesterol esters. The cholesterol derivatives may also be substituted cholesterol or substituted cholesterol esters. The complex of the present disclosure may also contain oxidized sterols, such as but not limited to, oxidized cholesterol or oxidized sterol derivatives (such as but not limited to, oxidized cholesterol esters). In some embodiments, the complex does not contain cholesterol and / or its derivatives (cholesterol esters or oxidized cholesterol esters).

[0137] Detergent The complex may contain one or more detergents. The detergents may be zwitterionic, nonionic, cationic, anionic, or a combination thereof. Exemplary zwitterionic detergents include 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), 3-[(3-cholamidopropyl)dimethylammonio]-2-hydroxy-1-propanesulfonate (CHAPSO), and N,N-dimethyldodecylamine N-oxide (LDAO). Exemplary non-ionic detergents include D-(+)-trehalose 6-monooleate, N-octanoyl-N-methylglucamine, N-nonanoyl-N-methylglucamine, N-decanoyl-N-methylglucamine, 1-(7Z-hexadecenoyl)-rac-glycerol, 1-(8Z-hexadecenoyl)-rac-glycerol, 1-(8Z-heptadecenoyl)-rac-glycerol, 1-(9Z-hexadecenoyl)-rac-glycerol, 1-decanoyl-rac-glycerol. Exemplary cationic detergents include (S)-O-methyl-serine dodecylamide hydrochloride, dodecylammonium chloride, decyltrimethylammonium bromide, and cetyltrimethylammonium sulfate. Exemplary anionic detergents include cholesteryl hemisuccinate, cholate, alkyl sulfate, and alkyl sulfonate.

[0138] 5.1.4.3.Fatty acids The complex can contain one or more fatty acids, which can include short chain fatty acids having an aliphatic tail of 5 carbons or less (e.g., butyric acid, isobutyric acid, valeric acid, or isovaleric acid), medium chain fatty acids having an aliphatic tail of 6-12 carbons (e.g., caproic acid, caprylic acid, capric acid, or lauric acid), long chain fatty acids having an aliphatic tail of 13-21 carbons (e.g., myristic acid, palmitic acid, stearic acid, or arachidic acid), very long chain fatty acids having an aliphatic tail of 22 or more carbons (e.g., behenic acid, lignoceric acid, or cerotic acid), or combinations thereof. The one or more fatty acids may be saturated (e.g., caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, behenic acid, lignoceric acid, or cerotic acid), unsaturated (e.g., myristoleic acid, palmitoleic acid, sapienic acid, oleic acid, elaidic acid, vaccenic acid, linoleic acid, linoleelaidic acid, alpha-linolenic acid, arachidonic acid, eicosapentaenoic acid, erucic acid, or docosahexaenoic acid), or a combination thereof. The unsaturated fatty acid may be a cis or trans fatty acid. In some embodiments, the unsaturated fatty acid used in the conjugates of the present disclosure is a cis fatty acid.

[0139] 5.1.4.4. Nonpolar molecules and sterols attached to sugars The conjugates can contain one or more amphiphilic molecules that include a nonpolar molecule or moiety (e.g., a hydrocarbon chain, acyl, or diacyl chain) or a sterol (e.g., cholesterol) attached with a sugar (e.g., a monosaccharide such as glucose or galactose, or a disaccharide such as maltose or trehalose). The sugar can be a modified or substituted sugar. Exemplary amphiphilic molecules comprising apolar molecules attached to a sugar include dodecan-2-yloxy-β-D-maltoside, tridecan-3-yloxy-β-D-maltoside, tridecan-2-yloxy-β-D-maltoside, n-dodecyl-β-D-maltoside (DDM), n-octyl-β-D-glucoside, n-nonyl-β-D-glucoside, n-decyl-β-D-maltoside, n-dodecyl-β-D-maltopyranoside, 4-n-dodecyl-α,α-trehalose, 6-n-dodecyl-α,α-trehalose, and 3-n-dodecyl-α,α-trehalose.

[0140] In some embodiments, the non-polar moiety is an acyl or diacyl chain.

[0141] In some embodiments, the sugar is a modified or substituted sugar.

[0142] 5.1.5. Formulations Lipid-binding protein-based conjugates can be formulated for the intended route of administration, for example, according to techniques known in the art (e.g., as described in Allen et al., eds., 2012, Remington: The Science and Practice of Pharmacy, 22nd Edition, Pharmaceutical Press, London, UK).

[0143] CER-001 intended for administration by infusion can be formulated in a phosphate buffer with sucrose and mannitol excipients, for example as described in WO 2012 / 109162.

[0144] 5.2. Target population The subjects which can be treated in accordance with the methods described herein are preferably mammals, most preferably human beings.

[0145] In some embodiments, the subject has or is at risk of developing leukocytosis. Leukocytosis may be caused by various conditions, such as infection, inflammatory process, and primary bone marrow disorders (e.g., acute leukemia, chronic leukemia, and myeloproliferative disorders). Physical stress, for example, from epileptic seizures, anesthesia, or overexertion, and emotional stress may also raise the leukocytosis count. Leukocytosis may also be caused by drugs, for example, corticosteroids, lithium, and beta-agonists. Increases in eosinophil or basophil counts resulting from various infections, allergic reactions, and other causes may also lead to leukocytosis in some cases.

[0146] In some embodiments, the subject with leukocytosis is 11×10 9 pcs / L higher and / or 100×10 9 In some embodiments, the subject has a white blood cell count of less than 11×10 9 pcs / L~50×10 9 pcs / L, 11×10 9 pcs / L~25×10 9 pcs / L, 25×10 9 pcs / L~50×10 9 pcs / L or 50 x 10 9 pcs / L~100×10 9 The patient had a white blood cell count of 10 / L.

[0147] In some embodiments, the subject is 9 Higher than 12 x 10 pieces / L 9 Higher than 15 x 10 pieces / L 9 pcs / L or higher, or 20 x 10 9 Alternatively or additionally, in some embodiments, the subject has a white blood cell count of greater than 100×10 9 Less than 50 x 10 pieces / L 9 Less than 25 x 10 9Less than 15 x 10 9 Have a white blood cell count of less than cells / L.

[0148] In some embodiments, the subject is 9 pcs / L~15×10 9 pcs / L or 15 x 10 9 pcs / L~20×10 9 The patient had a white blood cell count of 10 / L.

[0149] The subject may have one or more symptoms associated with leukocytosis. Exemplary symptoms associated with leukocytosis include fever, bleeding or bruising, sweating, pain or tingling in the legs, arms, or abdomen, vision problems (e.g., blurred vision, double vision, blind spots, cloudy vision, dim vision, or a combination thereof), unclear thinking, loss of appetite, or difficulty breathing (e.g., shortness of breath, lower than normal blood oxygen levels).

[0150] In some embodiments, the subject has or is at risk of developing leukocytosis due to inflammation, infection, white blood cell disorder, physical stress, emotional stress, medication, or an allergic reaction.

[0151] In some embodiments, the subject having or at risk of developing leukocytosis has an infection, e.g., a bacterial, fungal, parasitic, or viral infection (e.g., a coronavirus infection such as COVID-19 or influenza).

[0152] In some embodiments, the subject having or at risk of developing leukocytosis has diabetes.

[0153] In some embodiments, a subject having or at risk of developing leukocytosis has received or is receiving a drug that has leukocytosis as a side effect, such as a steroid, a corticosteroid, lithium, or a beta-agonist.

[0154] In some embodiments, the subject having or at risk of developing leukocytosis has a white blood cell disorder.

[0155] In some embodiments, the leukocyte disorder is a primary bone marrow disorder (eg, acute leukemia, chronic leukemia, or a myeloproliferative disorder).

[0156] In some embodiments, the white blood cell disorder is neutrophilia (eg, idiopathic neutrophilia, in some embodiments, chronic idiopathic neutrophilia).

[0157] In some embodiments, the white blood cell disorder is hemolytic anemia.

[0158] In some embodiments, the white blood cell disorder is thrombocytopenia (eg, idiopathic thrombocytopenia or essential thrombocytopenia).

[0159] In some embodiments, the white blood cell disorder is cancer (eg, bone cancer or blood cancer).

[0160] In some embodiments, the white blood cell disorder is a myeloproliferative disorder.

[0161] In some embodiments, the white blood cell disorder is a blood cancer.

[0162] In some embodiments, the white blood cell disorder is lymphoma.

[0163] In some embodiments, the leukocyte disorder is leukemia (eg, lymphocytic leukemia, chronic eosinophilic leukemia, chronic myeloid leukemia, or chronic neutrophilic leukemia).

[0164] In some embodiments, the white blood cell disorder is polycythemia vera.

[0165] In some embodiments, the leukocyte disorder is myelofibrosis, for example primary myelofibrosis.

[0166] In some embodiments of the methods of treating a subject having or at risk of developing leukocytosis and / or the methods of treating a subject having one or more symptoms associated with leukocytosis, administration of a lipid binding protein-based conjugate is effective to reduce the subject's white blood cell count and / or ameliorate one or more symptoms associated with leukocytosis.

[0167] In some embodiments, the subject has endothelial dysfunction (e.g., a subject experiencing or having experienced an acute coronary syndrome or stroke). Examples of acute coronary syndromes include myocardial infarction (e.g., ST-segment elevation myocardial infarction, non-ST-segment elevation myocardial infarction) and unstable angina.

[0168] In some embodiments, the subject is experiencing an acute coronary syndrome. In other embodiments, the subject has experienced an acute coronary syndrome. In other embodiments, the subject has experienced a stroke. In other embodiments, the subject has experienced a stroke.

[0169] In some aspects, the subject has or is at risk of developing carditis (e.g., myocarditis and / or pericarditis). In some embodiments, the subject is male. In some embodiments, the subject is under 60 years old, under 50 years old, under 40 years old, under 30 years old, under 20 years old, and / or at least 5 years old, at least 8 years old, at least 10 years old, at least 12 years old, at least 15 years old, or at least 18 years old.

[0170] In some embodiments, the subject having or at risk of developing carditis has an infection, e.g., a viral infection or a bacterial infection. Exemplary viral infections include influenza and coronavirus infection. In some embodiments, the subject has a COVID-19 infection. COVID-19 vaccines, e.g., mRNA COVID-19 vaccines, have been associated with carditis, particularly in young men. Thus, in some embodiments, the subject at risk of developing carditis is a young man (e.g., at least 5, 8, 10, 12, 15, or 18 years old, and less than 30 or 20 years old). In some embodiments, the subject receives a vaccine (e.g., a COVID-19 vaccine) prior to (e.g., 1-10 days) administration of the lipid-binding protein-based complex. In other embodiments, the subject receives a vaccine (e.g., a COVID-19 vaccine) after (e.g., 1-10 days) administration of the lipid-binding protein-based complex. In other embodiments, the subject receives a vaccine (e.g., a COVID-19 vaccine) concurrently (e.g., on the same day) with administration of the lipid-binding protein-based complex.

[0171] In some embodiments of the methods of the present disclosure, the subject has a SOFA score of 1 to 4, e.g., a score of 1, 2, 3, or 4, prior to treatment with the lipid-binding protein-based conjugate (see Vincent et al. 1996, Intensive Care Med, 22:707-710).

[0172] In some embodiments of the methods of the present disclosure, the subject has an endotoxin activity level measured by Endotoxin Activity Assay (EEA™) (Spectral Medical) of >0.6 prior to administration of the lipid-binding protein-based conjugate (see Marshall et al., 2004, J Infect Dis. 190(3):527-34).

[0173] In another aspect, the subject is a subject in need of a reduction in serum levels of one or more inflammatory markers, for example, a subject with elevated levels of one or more inflammatory markers compared to normal levels. Exemplary inflammatory cytokines include interleukin 6 (IL-6), C-reactive protein, D-dimer, ferritin, interleukin 8 (IL-8), granulocyte-macrophage colony stimulating factor (GM-CSF), monocyte chemoattractant protein (MCP) 1, and tumor necrosis factor alpha (TNFα). In some embodiments, the one or more cytokines include IL-6. In some embodiments, the one or more cytokines include a combination of the foregoing, for example, two, three, four, five, six, seven, or all eight of interleukin 6 (IL-6), C-reactive protein, D-dimer, ferritin, interleukin 8 (IL-8), granulocyte-macrophage colony stimulating factor (GM-CSF), monocyte chemoattractant protein (MCP) 1, and tumor necrosis factor alpha (TNFα).

[0174] 5.3. Medication Regimen The methods of the disclosure typically involve multiple administrations of a lipid-binding protein-based conjugate (e.g., CER-001), e.g., 2-20 individual doses (e.g., 2-16, 2-12, 2-10, 2-8, 2-6, 2-4, 4-20, 4-16, 4-10, 4-6, 6-20, 6-16, 6-10, 6-8, 8-20, 8-16, 8-10, 10-20, 10-16, or 16-20 individual doses). In some embodiments, the administration regimen may include 4 or more doses of a lipid-binding protein-based conjugate (e.g., CER-001), e.g., 5, 6, 7, 8, 9, 10, 11, 12, or more than 12 doses.

[0175] In some embodiments, the lipid-binding protein-based complex is administered according to the induction and optionally consolidation therapy regimen described in sections 5.3.1 and 5.3.2, respectively. In some embodiments, the lipid-binding protein-based complex can be administered in a single stage, for example according to the administration regimen described in this section. In some embodiments, the subject is not treated with the lipid-binding protein-based complex according to a maintenance regimen, for example a regimen that includes administration of the lipid-binding protein-based complex for a long period of time (e.g., for more than one month).

[0176] The lipid binding protein-based conjugate (eg, CER-001) dosing regimen of the present disclosure can last up to one week, one week, or more than one week (eg, two weeks).

[0177] For example, a lipid binding protein-based conjugate (e.g., CER-001) administration regimen can include administering: - CER-001, five doses over one week, - CER-001, six doses over one week, - CER-001, seven doses over one week, - 10 doses of CER-001 over 2 weeks, - 12 doses of CER-001 over 2 weeks, - 14 doses of CER-001 over 2 weeks.

[0178] In one embodiment, the method of the disclosure involves administering seven doses of CER-001 over a one week period, for example, on days 1, 2, 3, 4, 5, 6, and 7.

[0179] In some embodiments of the disclosed methods, the lipid-binding protein-based conjugate (e.g., CER-001) is administered daily, e.g., daily for at least 5 days, at least 6 days, at least 7 days, or for more than 7 days (e.g., daily for up to 1 week or daily for up to 2 weeks). In some embodiments of the disclosed methods, the lipid-binding protein-based conjugate (e.g., CER-001) is administered in multiple doses approximately 12 hours apart (e.g., 4-6 doses administered approximately 12 hours apart). In other embodiments, the lipid-binding protein-based conjugate (e.g., CER-001) is administered less frequently, e.g., every other day, twice a week, three times a week, or once a week.

[0180] In practice, a dosing window can be provided to accommodate slight variations in dosing schedule, for example, multiple weekly dosing, e.g., a window of ±2 days or ±1 day around the dosage date can be used.

[0181] The lipid-binding protein-based conjugate (e.g., CER-001) can be administered in the disclosed method for a predetermined period of time, for example, for one week. Alternatively, administration of the lipid-binding protein-based conjugate (e.g., CER-001) can be continued until one or more symptoms of the condition are reduced, or until the serum level of one or more inflammatory markers is reduced, for example, reduced to normal levels, or reduced compared to the subject's baseline value, for example, the baseline value measured before the start of lipid-binding protein-based conjugate (e.g., CER-001) treatment. Reference or "normal" levels of various inflammatory markers are known in the art. For example, the Mayo Clinic Laboratories test catalog (www.mayocliniclabs.com / test-catalog) provides the following reference values: IL-6: ≦1.8 pg / mL, C-reactive protein: ≦8.0 mg / mL, D-dimer: ≦500 ng / mL fibrinogen equivalent units (FEU), ferritin: 24-336 mcg / L (men), 11-307 mcg / L (women), IL-8 <57.8 pg / mL, TNF-α <5.6 pg / mL.

[0182] The disclosed methods (e.g., methods of treating a condition or symptom described herein) typically include administering a high dose of a lipid-binding protein-based conjugate (e.g., CER-001). The high dose can be a collection of multiple individual doses (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more than 10 individual doses), administered, for example, over multiple days (e.g., a period of 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days). In some embodiments, the individual doses of the high dose are administered daily, twice daily (e.g., approximately 12 hours apart), or 2-3 days apart.

[0183] In some embodiments, the high dose is effective to ameliorate one or more symptoms associated with leukocytosis.

[0184] In some embodiments, a high dose is an amount effective to increase a subject's HDL and / or ApoA-I blood levels and / or improve a subject's vascular endothelial function, e.g., as measured by circulating vascular cell adhesion molecule 1 (VCAM-1) and / or intercellular adhesion molecule 1 (ICAM-1) levels. In some embodiments, a high dose or individual dose is an amount that increases a subject's HDL and / or ApoA-I levels by at least 25%, at least 30%, or at least 35% 2-4 hours after administration.

[0185] In embodiments, the high dose is effective to reduce a subject's circulating VCAM-1 to at least 100ng / mL, at least 200ng / mL, at least 300ng / mL, and / or 400ng / mL within 2 days of the first administration of the lipid-binding protein-based complex. In embodiments, the high dose is effective to reduce a subject's circulating VCAM-1 to at least 100ng / mL, at least 200ng / mL, at least 300ng / mL, and / or 400ng / mL within 5 days of the first administration of the lipid-binding protein-based complex.

[0186] In embodiments, the high dose is effective to reduce a subject's circulating ICAM-1 to at least 50 ng / mL, at least 75 ng / mL, at least 100 ng / mL, and / or to 100 ng / mL or 125 ng / mL within 2 days of the first administration of the lipid binding protein-based complex. In embodiments, the high dose is effective to reduce a subject's circulating ICAM-1 to at least 50 ng / mL, at least 75 ng / mL, at least 100 ng / mL, and / or to 125 ng / mL within 5 days of the first administration of the lipid binding protein-based complex.

[0187] In some embodiments, the high dose is an amount effective to reduce serum levels of one or more inflammatory markers, such as one or more of IL-6, C-reactive protein, D-dimer, ferritin, IL-8, GM-CSF, MCP1, and TNF-α. In some embodiments, the serum levels of one or more inflammatory markers are reduced from an elevated range to the normal range and / or are reduced by at least 20%, at least 40%, or at least 60%.

[0188] In some embodiments, the high dose is effective to reduce serum levels of IL-6 by at least 20 pg / mL, at least 30 pg / mL, or at least 40 pg / mL, and / or to 40 pg / mL within 2 days of the first administration of the lipid binding protein-based complex. In some embodiments, the high dose is effective to reduce serum levels of IL-6 by at least 20 pg / mL, at least 30 pg / mL, or at least 40 pg / mL, and / or to 50 pg / mL or to 60 pg / mL within 5 days of the first administration of the lipid binding protein-based complex.

[0189] In some embodiments, the high dose is effective to reduce serum levels of C-reactive protein.

[0190] In some embodiments, the high dose is effective to reduce serum levels of D-dimers.

[0191] In some embodiments, the high dose is effective for reducing serum levels of ferritin. In some embodiments, the high dose is effective for reducing serum levels of ferritin to at least 200ng / mL, at least 300ng / mL, or at least 400ng / mL, and / or 700ng / mL within 2 days of the first administration of the lipid-binding protein-based complex. In some embodiments, the high dose is effective for reducing serum levels of ferritin to at least 200ng / mL, at least 300ng / mL, or at least 400ng / mL, and / or 700ng / mL within 5 days of the first administration of the lipid-binding protein-based complex.

[0192] In some embodiments, the high dose is effective to reduce serum levels of interleukin 8 (IL-8). In some embodiments, the high dose is effective to reduce serum levels of IL-8 to at least 100 pg / mL, at least 150 pg / mL, and / or 300 pg / mL within 2 days of the first administration of the lipid-binding protein-based complex. In some embodiments, the high dose is effective to reduce serum levels of IL-8 to at least 100 pg / mL, at least 150 pg / mL, and / or 300 pg / mL within 5 days of the first administration of the lipid-binding protein-based complex.

[0193] In some embodiments, the high dose is effective to reduce serum levels of monocyte chemoattractant protein (MCP) 1 and / or tumor necrosis factor alpha (TNF-α) and / or KIM-1.

[0194] In some embodiments, the high dose is effective to reduce the white blood cell count of the subject. In some embodiments, the high dose is effective to reduce the white blood cell count of the subject to at least 2000 WBC / mL, at least 3000 WBC / mL, at least 4000 WBC / mL, at least 5000 WBC / mL, and / or 8000 WBC / mL within 2 days of the first administration of the lipid-binding protein-based complex. In some embodiments, the high dose is effective to reduce the white blood cell count of the subject to at least 2000 WBC / mL, at least 3000 WBC / mL, at least 4000 WBC / mL, at least 5000 WBC / mL, and / or 6000 WBC / mL within 5 days of the first administration of the lipid-binding protein-based complex.

[0195] In some embodiments, the high dose is effective to transiently increase serum triglyceride levels, for example, for up to 9 days.

[0196] In some embodiments, the high dose is a dose effective to prevent carditis in a subject at risk for carditis or to reduce the severity of carditis in a subject having or at risk for carditis.

[0197] The dose of lipid-binding protein-based complex (e.g., CER-001) administered to a subject (e.g., an individual dose that forms a higher dose when aggregated with one or more other individual doses) may, in some embodiments, be in the range of 4-40 mg / kg (e.g., 10-40 mg / kg) on ​​a protein weight basis (e.g., 5, 10, 15, 20, 25, 30, 35, or 40 mg / kg, or any range bounded by any two of the aforementioned values, e.g., 10-20 mg / kg, 15-25 mg / kg, 20-40 mg / kg, 25-35 mg / kg, or 30-40 mg / kg). As used herein, the expression "protein weight basis" means that the dose of lipid-binding protein-based complex (e.g., CER-001) administered to a subject is calculated based on the amount of ApoA-I in the lipid-binding protein-based complex (e.g., CER-001) administered and the body weight of the subject. For example, a subject weighing 70 kg and receiving a dose of CER-001 of 20 mg / kg would receive an amount of CER-001 that provides 1400 mg of ApoA-I (70 kg x 20 mg / kg).

[0198] In yet other aspects, lipid-binding protein-based conjugates (e.g., CER-001) can be administered on a unit dosage basis. Unit dosages used in the methods of the present disclosure may, in some embodiments, range from 300 mg to 4000 mg (e.g., 600 mg to 4000 mg) per administration (based on protein weight).

[0199] In certain embodiments, the dosage of the lipid binding protein-based conjugate (e.g., CER-001) is between 600 mg and 3000 mg, between 800 mg and 3000 mg, between 1000 mg and 2400 mg, or between 1000 mg and 2000 mg per dose (based on protein weight).

[0200] In some aspects, a high dose of a lipid-binding protein-based conjugate (e.g., CER-001), e.g., a collection of multiple individual doses, is between 600 mg and 40 g (by protein weight). In certain embodiments, the high dose is between 3 g and 35 g or between 5 g and 30 g (by protein weight).

[0201] The lipid-binding protein-based conjugate (e.g., CER-001) is preferably administered as an IV infusion. For example, a stock solution of CER-001 can be diluted with standard saline, such as saline (0.9% NaCl), to a total volume of 125-250 ml. In some embodiments, subjects weighing less than 80 kg will receive a total volume of 125 ml, while subjects weighing at least 80 kg will receive a total volume of 250 ml. In some embodiments, a dose of CER-001 is administered in a total volume of 250 ml. The lipid-binding protein-based conjugate (e.g., CER-001) can be administered over a period ranging from 1 hour to 24 hours. Depending on the needs of the subject, administration can be by slow infusion for a period of more than 1 hour (e.g., up to 2 hours or up to 24 hours), by rapid infusion for up to 1 hour, or by a single bolus injection. In one embodiment, the lipid-binding protein-based conjugate (e.g., CER-001) is administered using a fixed rate infusion pump, e.g., 125 ml / hr or 250 ml / hr, over a period of 1 hour. In one embodiment, the lipid-binding protein-based conjugate (e.g., CER-001) dose is administered as an infusion over a period of 24 hours.

[0202] 5.3.1. Induction regimen In one embodiment, an induction regimen suitable for use in the methods of the present disclosure involves administration of multiple doses of a lipid-binding protein-based conjugate (e.g., CER-001) over multiple consecutive days, e.g., three consecutive days.

[0203] In some embodiments, an induction regimen suitable for use in the methods of the present disclosure involves administration of a lipid-binding protein-based conjugate (e.g., CER-001) twice daily, e.g., twice daily administration for consecutive days. Twice daily administration can include, for example, two doses approximately 12 hours apart, or a morning dose and an evening dose, which can be more or less than 12 hours apart.

[0204] In one embodiment, the induction regimen comprises two doses per day of a lipid-binding protein-based conjugate (eg, CER-001) for three consecutive days.

[0205] The therapeutic dose of lipid-binding protein-based conjugate (e.g., CER-001) administered by infusion in an induction regimen can be in the range of 4-40 mg / kg (e.g., 4-30 mg / kg) on ​​a protein weight basis (e.g., 4, 5, 6, 7, 8, 9, 10, 12, 15, 20, 25, 30, or 40 mg / kg, or any range bounded by any two of the preceding values, e.g., 5-15 mg / kg, 10-20 mg / kg, or 15-25 mg / kg). In some embodiments, the dose of lipid-binding protein-based conjugate (e.g., CER-001) used in an induction regimen is 5 mg / kg. In some embodiments, the dose of lipid-binding protein-based conjugate (e.g., CER-001) used in an induction regimen is 10 mg / kg. In some embodiments, the dose of lipid-binding protein-based conjugate (e.g., CER-001) used in an induction regimen is 15 mg / kg. In some embodiments, the dose of lipid-binding protein-based conjugate (e.g., CER-001) used in the induction regimen is 20 mg / kg. In some embodiments, the induction regimen includes 6 doses of lipid-binding protein-based conjugate (e.g., CER-001) administered over 3 days at doses of 5 mg / kg, 10 mg / kg, 15 mg / kg, or 20 mg / kg.

[0206] In yet another embodiment, the lipid-binding protein-based conjugate (e.g., CER-001) can be administered on a unit dosage basis. The unit dosage used in the induction phase can vary from 300 mg to 4000 mg (e.g., 300 mg to 3000 mg) (protein weight basis) per infusion.

[0207] In certain embodiments, the dosage of lipid-binding protein-based conjugate (e.g., CER-001) used during the induction phase is 300 mg to 1500 mg, 400 mg to 1500 mg, 500 mg to 1200 mg, or 500 mg to 1000 mg (protein weight basis) per infusion.

[0208] 5.3.2. Consolidation regimens Consolidation therapy regimens suitable for use in the methods of the present disclosure involve administration of one or more doses of a lipid-binding protein-based conjugate (eg, CER-001) following an induction regimen.

[0209] In one embodiment, the consolidation therapy regimen includes administering two doses of a lipid-binding protein-based conjugate (e.g., CER-001), for example, the two doses can be administered approximately 12 hours apart, or as a morning dose and an evening dose, which can be more or less than 12 hours apart.

[0210] A dose of a lipid-binding protein-based conjugate (e.g., CER-001) in a consolidation therapy regimen can be administered on day 6 of a dosing regimen starting from day 1 of an induction regimen in some embodiments. A dose of a lipid-binding protein-based conjugate (e.g., CER-001) in a consolidation therapy regimen can be administered on day 4 of a dosing regimen starting from day 1 of an induction regimen in some embodiments. A dose of a lipid-binding protein-based conjugate (e.g., CER-001) in a consolidation therapy regimen can be administered on day 5 of a dosing regimen starting from day 1 of an induction regimen in some embodiments. A dose of a lipid-binding protein-based conjugate (e.g., CER-001) in a consolidation therapy regimen can be administered on day 7 of a dosing regimen starting from day 1 of an induction regimen in some embodiments.

[0211] The therapeutic dose of lipid-binding protein-based conjugate (e.g., CER-001) administered by infusion in a consolidation therapy regimen can be in the range of 4 mg / kg to 40 mg / kg (e.g., 4 to 30 mg / kg) on ​​a protein weight basis (e.g., 4, 5, 6, 7, 8, 9, 10, 12, 15, 20, 25, 30, or 40 mg / kg, or any range bounded by any two of the preceding values, e.g., 5 to 15 mg / kg, 10 to 20 mg / kg, or 15 to 25 mg / kg). In some embodiments, the dose of lipid-binding protein-based conjugate (e.g., CER-001) used in a consolidation therapy regimen is 5 mg / kg. In some embodiments, the dose of lipid-binding protein-based conjugate (e.g., CER-001) used in a consolidation therapy regimen is 10 mg / kg. In some embodiments, the dose of lipid-binding protein-based conjugate (e.g., CER-001) in a consolidation therapy regimen is 15 mg / kg. In some embodiments, the dose of lipid-binding protein-based conjugate (e.g., CER-001) used in the consolidation therapy regimen is 20 mg / kg. In some embodiments, the consolidation therapy regimen includes two doses of lipid-binding protein-based conjugate (e.g., CER-001) administered daily at doses of 5 mg / kg, 10 mg / kg, 15 mg / kg, or 20 mg / kg.

[0212] In yet another embodiment, the lipid-binding protein-based conjugate (e.g., CER-001) can be administered on a unit dosage basis. The unit dosage used in the consolidation phase can vary from 300 mg to 4000 mg (e.g., 300 mg to 3000 mg) (protein weight basis) per infusion.

[0213] In certain embodiments, the dosage of lipid-binding protein-based conjugate (e.g., CER-001) used during the consolidation phase is 300 mg to 1500 mg, 400 mg to 1500 mg, 500 mg to 1200 mg, or 500 mg to 1000 mg (protein weight basis) per infusion.

[0214] The lipid-binding protein-based conjugate (eg, CER-001) can be administered during the consolidation phase in the same manner as described in Section 5.3, for example, as an IV infusion over a one hour period.

[0215] 5.4. Combination Therapy The lipid-binding protein-based complex (e.g., CER-001) can be administered to the subject described herein as a monotherapy or as part of a combination therapy regimen. For example, the combination therapy can include a lipid-binding protein-based complex (e.g., CER-001) in combination with a standard treatment for sepsis or other infections. See, for example, Rhodes et al., 2017, Intensive Care Med 43:304-377; Dugar et al., 2020, Cleveland Clinic Journal of Medicine 87(1):53-64.

[0216] In some embodiments, the subject is treated with a lipid-binding protein-based conjugate (e.g., CER-001) in combination with fluid replacement therapy. In some embodiments, the subject is treated with a lipid-binding protein-based conjugate (e.g., CER-001) in combination with an antimicrobial. In some embodiments, the subject is treated with a lipid-binding protein-based conjugate (e.g., CER-001) in combination with an antibiotic (e.g., ceftriaxone, meropenem, ceftazidime, cefotaxime, cefepime, piperacillin and tazobactam, ampicillin and sulbactam, imipenem and cilastatin, levofloxacin, or clindamycin). In some embodiments, the subject is treated with a lipid-binding protein-based conjugate (e.g., CER-001) in combination with an antiviral agent. In some embodiments, the subject is treated with a lipid-binding protein-based conjugate (e.g., CER-001) in combination with an agent that increases blood pressure (e.g., norepinephrine or epinephrine).

[0217] The combination therapy regimen may, in some embodiments, include one or more anti-IL-6 agents and / or one or more other agents for treating CRS, such as corticosteroids (e.g., methylprednisolone and / or dexamethasone). Exemplary anti-IL6 agents include tocilizumab, siltuximab, olokizumab, elcilimomab, BMS-945429, sirukumab, revilimab, and CPSI-2364. In some embodiments, a lipid-binding protein-based conjugate (e.g., CER-001) is administered in combination with tocilizumab. Subjects who have or have had a COVID-19 infection can be treated with the lipid-binding protein-based conjugate (e.g., CER-001) in combination with one or more additional therapies, such as antibodies from recovered COVID-19 patients, antibodies against the COVID-19 spike protein, one or more antiviral agents (e.g., lopinavir, remdesivir, danoprevir, galidesivir, darunavir, ritonavir), chloroquine, hydroxychloroquine, azithromycin, interferon (e.g., interferon alpha or interferon beta, each of which may be pegylated), or combinations thereof.

[0218] In certain embodiments, an antihistamine (e.g., diphenhydramine, cetirizine, fexofenadine, or loratadine) can be administered prior to administration of the lipid-binding protein-based conjugate (e.g., CER-001). The antihistamine can reduce the likelihood of an allergic reaction. EXAMPLES

[0219] 6. Working Example [Example 1] 6.1. Example 1: CER-001 treatment in a porcine model of LPS-induced AKI The ability of CER-001 to attenuate sepsis-associated AKI was evaluated in a porcine model of lipopolysaccharide (LPS)-induced AKI.

[0220] Materials and Methods Pigs were randomized into three groups: LPS (endotoxemic pigs, n=3), single dose CER-001 treated pigs (endotoxemic pigs treated with a single dose of 20 mg / kg CER-001, n=3), and multiple dose CER-001 treated pigs (endotoxemic pigs treated with two doses of 20 mg / kg CER-001, n=3).

[0221] Sepsis was induced in pigs by intravenous infusion of saline containing 300 μg / kg LPS at T0. Pigs treated with single dose CER-001 and pigs treated with multiple doses of CER-001 received a 20 mg / kg dose of CER-001 at T0. Pigs treated with multiple doses of CER-001 received a second 20 mg / kg dose of CER-001 3 hours later (T3). Serum IL-6, LPS, MCP-1, sVCAM-1, and sICAM-1 levels were monitored over time. Renal tissue damage and fibrosis were evaluated at the end of the study period.

[0222] 6.1.2.Results Increased survival was observed in both CER-001-treated groups compared to the LPS group (data not shown). LPS injection resulted in a time-dependent increase in IL-6 in endotoxemic animals compared to basal conditions (T0) (Figure 1). CER-001 treatment was able to reverse the LPS effect, as shown by the reduced IL-6 levels (Figure 1, "20MG" and "40MG"). A second infusion of CER-001 3 hours (T3) after the first dose potently reduced IL-6 serum levels to basal levels by the end of the study (Tend) (Figure 1, "40MG"). Similarly, while higher levels of MCP-1 were observed in endotoxemic pigs compared to basal conditions, MCP-1 levels were lower in pigs treated with CER-001 (data not shown).

[0223] Endothelial dysfunction was assessed by measuring sVCAM-1 and sICAM-1 serum levels. A time-dependent increase in sVCAM-1 and sICAM-1 was observed in endotoxemic animals, while CER-001 treatment potently reduced sVCAM-1 and sICAM-1 levels in both treatment groups (Figures 2 and 3, respectively). Consistent with the IL-6 results, infusion of two doses of CER-001 (Figure 2, "40MG") was more efficient in reducing sVCAM-1 to basal levels. LPS levels were potently reduced in animals treated with CER-001 (Figure 4, "20MG" and "40MG"), with the effect being more evident after the second infusion of CER-001 (Figure 4, "40MG").

[0224] Endotoxemic kidney biopsies presented tubular vacuolation, epithelial flattening, and some apoptotic tubular cells. CER-001 treatment significantly reduced the inflammatory process and tubular injury. In endotoxemic animals, Masson's trichrome staining revealed extensive collagen deposition at the interstitial level. In both CER-001-treated groups, there was significantly less collagen deposition in the renal parenchyma compared to the LPS group.

[0225] This preclinical data indicates that CER-001 treatment reduces systemic inflammation and endothelial dysfunction, thereby limiting renal injury in a porcine model of LPS-induced AKI.

[0226] [Example 2] 6.2. Example 2: A randomized pilot study comparing short-term CER-001 infusions at various doses to prevent sepsis-induced acute kidney injury Currently, there is no approved treatment for sepsis-associated AKI. Given that the inflammatory response to endotoxemia is the primary cause of hemodynamic instability and progression to AKI in septic patients, the primary objective of this study is to investigate whether the use of various doses of CER-001 in combination with standard of care (SOC) treatment is safe and effective, providing a new strategy to treat septic patients, reducing the inflammatory response, and preventing progression to AKI. Without being bound by theory, the predicted mechanism of action is two-fold and involves both binding of endotoxin by CER-001 and a direct anti-inflammatory effect of CER-001.

[0227] Study protocol Study Population: This is a single-center, randomized, dose-ranging (Phase II) study including patients with sepsis due to intra-abdominal infection or urosepsis admitted to the intensive care units (ICUs) of the participating centers. The investigators will ensure that all patients who meet the following inclusion and exclusion criteria will be offered enrollment in the study.

[0228] Inclusion Criteria: - Adult males or non-pregnant females ≥ 18 years of age at the time of enrollment, - Meeting criteria for sepsis 3, defined as an acute increase of at least 2 points in the SOFA score compared to the SOFA score at admission, - endotoxin levels (measured by Endotoxin Activity Assay (EEA™), Spectral Medical) >0.6 (see Marshall et al., 2004, J Infect Dis. 190(3):527-34), - Signed and dated informed consent by the patient or their legal representative.

[0229] Exclusion criteria: - Patients weighing more than 100 kg, -Alanine transaminase / aspartate transaminase (ALT / AST) > 5x upper limit of normal; - Stage 4 severe chronic kidney disease or requiring dialysis (i.e. estimated glomerular filtration rate (eGFR) < 30 ml / min / 1.73 m 2 ), - White blood cell count <2.0×10^9, - Pregnancy or breastfeeding, - Received an organ transplant within the past year - Anticipated transfer to another hospital outside the study site within 72 hours, - End-stage disease, including metastases or hematologic malignancies, with a life expectancy of less than 30 days (as assessed by the treating physician) or classified as "refusal to resuscitate"; - History of end-stage chronic organ failure, -HIV diagnosis, - Uncontrollable bleeding within the past 24 hours, - Patients who have used an investigational drug or device within 30 days of the first dose of CER-001.

[0230] Number of subjects: 20 subjects were enrolled and randomized (1:1:1:1) into 4 experimental groups: Group A patients continued to receive conventional treatment, Group B: patients added CER-001 at 5 mg / kg BID for 3 days followed by 5 mg / kg BID on day 6 to conventional treatment, Group C: patients added CER-001 at 10 mg / kg BID for 3 days followed by 10 mg / kg BID on day 6 to conventional treatment, and Group D patients added CER-001 at 20 mg / kg BID for 3 days followed by 20 mg / kg BID on day 6 to conventional treatment (Figure 5).

[0231] Duration of the Study: The study will be completed in 24 weeks (6 months). The enrollment period will be approximately 20 weeks (5 months) from the first subject enrolled. The end of the study will be the last visit of the last subject.

[0232] Primary Endpoint: The primary endpoint of the study is to define the safety and optimal dose of CER-001 in combination with standard of care in patients with sepsis sustained by Gram-negative bacteria.

[0233] Secondary Endpoints: The secondary endpoints were: - Change in endotoxin and IL-6 levels from baseline to days 3, 6, and 9. - Baseline is defined as the last measurement taken prior to dosing on Day 1. - Change in SOFA score from baseline to days 3, 6, and 9 (Vincent et al. 1996, Intensive Care Med, 22:707-710). - Changes in key inflammatory markers (CRP, D-dimer, ferritin, IL-8, GM-CSF, MCP1, and TNF-α) from baseline to days 3, 6, and 9. - Changes in AKI biomarkers and the development of AKI according to the KDIGO (Kidney Disease Improving Global Outcomes) criteria (KDIGO Clinical Practice Guideline for Acute Kidney Injury. Kidney International Supplements 2012; 2: 1-138) -30-day mortality - Independent medical experts review outcome data during the trial.

[0234] Interventions / Exposures: Twenty patients fulfilling the eligibility criteria, who signed and dated an Ethics Committee (EC) approved informed consent form, were randomized and assigned in a (1:1:1:1) ratio to conventional treatment (group A), low dose CER-001 (group B) or medium dose CER-001 (group C) or high dose CER-001 (group D). Conventional treatment was modulated according to clinical status. All non-experimental treatments were allowed to be administered concomitantly during the patient's participation in the study: any medication taken by the patient, other than the study drug specified by the protocol, will be considered as a concomitant medication and will be recorded in the study records.

[0235] Each patient will be identified by a patient number at screening. Once assigned to a patient, patient numbers will not be reused. The investigators who generate and enroll the randomization list and allocation sequence will not participate in any part of the task. The blocked randomization list will be appropriately concealed to prevent attempts to subvert the randomization.

[0236] Treatment group: All patients will receive conventional treatment. The treatment group will receive additional treatment with the study drug, specifically: - Group A: Conventional treatment (i.e. antibiotic treatment and hemodynamic support depending on the patient's condition). - Group B: conventional therapy + CER-001 at 5 mg / kg BID for 3 consecutive days, followed by 5 mg / kg BID on the 6th day. - Group C: conventional therapy + CER-001 at 10 mg / kg BID for 3 consecutive days, followed by 10 mg / kg BID on the 6th day. - Group D: conventional therapy + CER-001 at 20 mg / kg BID for 3 consecutive days, followed by 20 mg / kg BID on the 6th day.

[0237] Patients will be pretreated with an antihistamine prior to each CER-001 dose (e.g., dexchlorphenylamine 5 mg or hydroxyzine 100 mg) to avoid any potential infusion reactions. Patients may discontinue or withdraw from study drug if any of the following occur:

[0238] Any drug-related adverse events or other reasons that, in the investigator's opinion, compromise the patient's participation in the trial or the interpretation of the trial data (e.g. serious intercurrent illness requiring additional care measures or preventing further medication), significant tolerability problems.

[0239] Upon discontinuation of study drug, the study site will document the reason for drug discontinuation, continue to follow the patient clinically, and make every attempt to restart study drug within 2 days of study drug discontinuation, unless otherwise contraindicated.

[0240] Reasons for withdrawal from study drug may include, but are not limited to: - At the request of the investigator, for safety reasons, such as serious adverse reactions, - For other reasons, such as investigator request, patient noncompliance, etc. - At the patient's request or for reasons of durability - For other reasons, such as patient request, withdrawal of informed consent, etc.

[0241] Discontinuation of study drug alone does not constitute discontinuation or withdrawal from the study. Patients will continue to be followed as if they had completed the treatment phase. Patients who discontinue study medication prematurely (e.g., before completion of the third dose) will undergo end of study evaluations whenever possible.

[0242] Statistical analysis: Comparisons between groups are performed using appropriate statistical tests: dichotomous variables (baseline characteristics, mortality, occurrence of AKI) are compared using chi-square or Fisher's exact test, continuous baseline characteristics by ANOVA or Kruskal-Wallis test, Student's test, or Mann-Whitney U test, as appropriate. Changes in inflammatory markers are compared between groups by ANOVA and presented graphically. The proportion of patients with AKI and mortality are calculated for each group. All analyses are performed using SPSS 12.0 for Windows. p<0.05 is considered statistically significant.

[0243] Procedures: The following procedures will be performed during the Screening Visit: After randomization, subjects will begin treatment within 2 business days. -Informed consent - Medical History: Includes recording of past and present illnesses and collection of subject demographic data (date of birth, sex, and race). - Physical examination with review of system, height and weight, BMI, and waist circumference -Vital signs (pulse, blood pressure, and oral, ear, axillary, or core temperature). -Review of inclusion / exclusion criteria. - Adverse events will be recorded starting from the time informed consent is obtained. - Previous medications will be collected from 4 weeks prior to the first dose of test article. All current medications will be recorded. - Complete Blood Count (CBC) - Includes white blood cell (WBC) differential, platelet count, red blood cell (RBC), hemoglobin (Hb), and hematocrit (Hct). -Fasting Chemistry Panel / Electrolytes: Includes sodium, potassium, chloride, blood urea nitrogen (BUN, or urea), serum creatinine, calculated clearance creatinine (CKD-EPI), glucose, calcium, phosphorus, total protein, uric acid, AST, ALT, γGT, ALP, total and direct bilirubin, albumin, total cholesterol, HDL, LDL, triglycerides, LDH, CPK, -ABG (to assess respiratory and / or metabolic disorders) -ApoA-I (for pharmacokinetic and pharmacodynamic evaluation) -Coagulation tests - include prothrombin time (PT) (expressed as international normalized ratio [INR]) and partial thromboplastin time (PTT). -Urinary analysis - includes specific gravity, pH, protein / albumin assessment, glucose, ketones, and hemoglobin / blood. - Microalbuminuria and proteinuria, g / 24 hours - Serum or urine pregnancy test (for women of childbearing potential) within 7 days prior to randomization. - Pharmacokinetic and pharmacodynamic assessments will include apoA-I and total cholesterol levels. - Endotoxin levels are measured using the EAA™ kit. AKI biomarkers (TIMP-2 and IGFBP-7) are measured using the Nephrocheck® kit. Inflammatory markers include: CRP, D-dimer, ferritin, IL-6, IL-8, GM-CSF, MCP1, and TNF-α.

[0244] In addition to the biospecimens collected for daily routine laboratory evaluation at the central laboratory, biospecimens will be collected for research purposes, including the following: - Two tubes of 5 ml serum -One tube of 3ml plasma - 30ml of urine

[0245] These samples will be used to evaluate additional inflammatory cytokines and urinary biomolecules to obtain a more comprehensive characterization of enrolled patients, to better evaluate their response to treatment, to provide further information in follow-up, and more importantly, to discover new potential biomarkers that may be useful for the early diagnosis of sepsis-induced AKI. Analysis will be performed by ELISA tests and protein arrays.

[0246] At the Treatment Visit (Treatment Period): The treatment period is defined as the start of treatment. Visits are scheduled on days 3, 6, and 9. A final visit is scheduled on day 30. The following procedures will be performed during the treatment visits: - Records of adverse events and concomitant medications -Review of pertinent laboratory information -Physical examination -Assess vital signs (pulse, blood pressure, and oral, ear, axillary, or core temperature) -Continuously record adverse events and concomitant medications - Complete Blood Count (CBC) - Includes white blood cell (WBC) differential, platelet count, red blood cell (RBC), hemoglobin (Hb), and hematocrit (Hct). -Fasting Chemistry Panel / Electrolytes: Includes sodium, potassium, chloride, blood urea nitrogen (BUN, or urea), serum creatinine, and calculated clearance creatinine (CKD-EPI); -Glucose, calcium, phosphorus, total protein, uric acid, AST, ALT, γGT, ALP, total and direct bilirubin, albumin, total cholesterol, HDL, LDL, triglycerides, LDH, CPK -ABG (to assess respiratory and / or metabolic disorders) -ApoA-I (for pharmacokinetic and pharmacodynamic evaluation) -Coagulation tests - include prothrombin time (PT) (expressed as international normalized ratio [INR]) and partial thromboplastin time (PTT). -Urinary analysis - includes specific gravity, pH, protein / albumin assessment, glucose, ketones, and hemoglobin / blood. - Microalbuminuria and proteinuria, g / 24 hours - Serum or urine pregnancy test (for women of childbearing potential) within 7 days prior to randomization. - Pharmacokinetic and pharmacodynamic assessments will include apoA-I and total cholesterol levels. - Endotoxin levels are measured using the EAA™ kit. AKI biomarkers (TIMP-2 and IGFBP-7) are measured using the Nephrocheck® kit. Inflammatory markers include: CRP, D-dimer, ferritin, IL-6, IL-8, GM-CSF, MCP1, and TNF-α.

[0247] In addition to the biospecimens collected for daily routine laboratory evaluation at the central laboratory, biospecimens will be collected for research purposes, including the following: - Two tubes of 5 ml serum -One tube of 3ml plasma - 30ml of urine.

[0248] Clinical scores include the SOFA score (Table 2) and the KDIGO criteria for AKI assessment and staging (Table 3). The individual components of each score are documented.

[0249] [Table 2]

[0250] [Table 3]

[0251] Table 4 provides a summary of the study protocol for this example.

[0252] [Table 4]

[0253] Safety Assessment: Safety assessment will be accomplished utilizing information collected from the following assessments: physical examination (including weight), vital signs (blood pressure, pulse, temperature), CBC differential, platelet count, blood chemistry, fasting lipid profile [including HDL-cholesterol, LDL-cholesterol, and lipoprotein(a)], urea, glucose, 24-hour urinary protein determination, serum creatinine and calculated creatinine clearance (CKD-EPI), and monitoring of adverse events. All women of childbearing potential will undergo a qualitative serum pregnancy test during pre-study screening / baseline assessment and thereafter if clinically indicated. Patients will be monitored and recorded for the occurrence of adverse events throughout the study. Adverse events volunteered by subjects or discovered by general questioning or physical examination by the investigator will be recorded. The duration (start and end dates), severity, cause, and relationship to study drug, patient outcome, actions taken, and an assessment of whether the event was serious will be recorded for each reported adverse event.

[0254] Adverse Events: Definitions The term "adverse event" is synonymous with the term "adverse experience" as used by the FDA. An adverse event (AE) is any untoward, undesirable, unscheduled clinical occurrence in the form of signs, symptoms, illness, or laboratory or physiological observations, occurring in humans participating in a clinical study, regardless of causality. This includes: - Any clinically significant worsening of a pre-existing condition. - Any recurrence of a pre-existing condition. - AE resulting from an overdose of the investigational test substance (i.e., a dose higher than that prescribed by a health care professional for clinical reasons), whether accidental or intentional. - AEs resulting from abuse of the investigational test substance (i.e., use without clinical justification). - AEs associated with discontinuation of use of the investigational test substance.

[0255] The procedure is not an AE, but the reason for the procedure may be an AE.

[0256] A "pre-existing condition" is a clinical condition that has been diagnosed and documented as part of the subject's medical history before the subject signs the informed consent form (including the condition being treated). Questions regarding whether the condition was present before the start of the active phase of the study and whether it has increased in severity and / or frequency are used to determine whether the event is a treatment-emergent adverse event (TEAE). An AE is considered treatment-emergent if (1) it is not present when the active phase of the study begins and is not a chronic condition that is part of the subject's medical history, or (2) it is present or part of the subject's medical history at the start of the active phase of the study but increases in severity or frequency during the active phase. The active phase of the study begins with the first dose of drug.

[0257] A "serious adverse event" is any AE occurring at any dose that meets one or more of the following criteria: -Brings death -It is life-threatening (see below) - Results in a qualifying hospitalization or a prolongation of an existing hospitalization (see below) -Causing persistent or significant disability or incapacity (see below) -Bringing about new malignancies -Resulting in birth abnormalities or birth defects.

[0258] Furthermore, a significant medical event that may not be fatal, life-threatening, or require hospitalization may be considered an SAE if, based on appropriate medical judgment, it may endanger the subject and require medical or surgical treatment to prevent one of the outcomes listed above. Examples of such events include allergic bronchospasm that requires intensive care in an emergency room or at home, blood dysbiosis or convulsions that do not require hospitalization, or the occurrence of drug dependence or abuse.

[0259] "Life-threatening adverse event" means any AE that, when it occurs, places a subject at risk of immediate death from the event. A life-threatening event does not include an event that could have caused death if it had occurred in a more severe form, but that did not pose an immediate risk of death when it did occur. For example, drug-induced hepatitis that resolves without evidence of liver failure is not considered life-threatening, even though drug-induced hepatitis of a more severe nature may be fatal.

[0260] Hospitalization or extended hospitalization is a criterion for considering an AE serious. In the absence of an AE, participating investigators should not report hospitalization or extended hospitalization on the form. This is the case in the following circumstances: Hospitalization or extended hospitalization is necessary for a procedure required by the protocol. Daytime or overnight study visits required by the protocol are not considered serious.

[0261] Timing of Reporting Serious Adverse Events: Any SAEs, regardless of causality, will be reported immediately (within 24 hours of the Investigator noticing the SAE) to the Medical Monitor by faxing a completed Serious Adverse Event Form. Follow-up information related to an SAE will be reported to the Medical Monitor (or designee) within 24 hours of receipt by the Investigator by faxing a completed Serious Adverse Event Form. Subjects will be closely observed and monitored until the condition resolves or stabilizes or its cause is identified. Any emergency situations will be reported immediately (within 24 hours) to the Medical Monitor (or designee) by contacting the Medical Monitor.

[0262] Reportable Events / Information: An AE or SAE, regardless of test article or protocol relatedness, may occur from the time the subject signs the informed consent form up to 15 days after the subject's last dose. This includes events occurring during screening and placebo run-in periods. Record all AEs and SAEs in source documentation and on the CRF. All AEs and SAEs occurring after the screening period will be recorded on the CRF.

[0263] For SEA: The Investigator will provide the Medical Monitor with all documentation related to the event (e.g., additional laboratory tests, medical examination reports, discharge summaries, post-mortem reports, etc.) in a timely manner. Subsequent progress reports on the subject will be provided to the Investigator until the event has subsided or, in the case of permanent impairment, the condition has stabilized.

[0264] The following events will be recorded and reported in the same timeframes and according to the same process as SAE:

[0265] Abuse and overdose (i.e., use for non-clinical reasons) of the test substance, with or without AEs. An overdose is a dose higher than that prescribed by a health care professional for clinical reasons. It is up to the participating investigator to determine whether a dose is an overdose.

[0266] Inadvertent or accidental exposure to the test substance, with or without an AE.

[0267] Post-study test article-related SAEs.

[0268] SAEs occurring after unauthorized or accidental use in people not participating in the study.

[0269] Any abnormal biological or vital sign values ​​deemed clinically relevant by the participating investigator. These will be reported in the same timeframe and following the same process as AEs or SAEs.

[0270] Recording and Reporting: At each required study visit, all AEs occurring since the previous visit will be recorded in the Adverse Event Record on the subject's CRF. Information recorded is based on the physical examination and any signs or symptoms detected during the subject's clinical evaluation. In addition to information obtained from these sources, subjects will be asked the following non-specific question: "How have you been feeling since your last visit?". Signs and symptoms will be recorded using standard medical terminology. The health outcome assessment survey administered to study subjects is intended to explore the subject's own perception of their quality of life. However, the investigator will review the survey for the presence of potential AEs or SAEs and will take the subject's perception into account when determining AE or SAE occurrence. Subject assessments are not intended to be influenced by the clinical investigator. Every effort will be made to maintain unbiased assessments. Include the following AE information (if applicable): specific condition or event and direction of change, whether the condition was pre-existing (i.e., history of an acute condition or chronic condition present at the start of the study) and, if so, whether it worsened (in severity and / or frequency), date and time of occurrence, severity, causal relationship to the test article, actions taken, and outcome. All laboratory abnormalities that, in the investigator's opinion, are clinically significant will be reported as AEs.

[0271] A causal relationship between an AE and the test article will be determined by the investigator based on the investigator's clinical judgment and the following definitions: - Definitely related: The event can be completely explained by the administration of the test substance. - Probably related: The event is more likely to be explained by the administration of the test article rather than the subject's clinical condition or another drug / treatment. -Potentially related: The event may be explained by administration of the test article or by the subject's clinical condition or other medications / treatments. -Probably not related: The event is more likely to be explained by the subject's clinical condition or another drug / treatment than by the test substance. - Definitely Not Related: The event can be completely explained by the subject's clinical condition or another drug / treatment.

[0272] When assessing the relationship between test substance administration and AEs, the following will be considered: - The temporal relationship between administration of the test substance and the AE -The biological plausibility of the relationship - Subject's underlying clinical condition or concomitant medications and / or treatments

[0273] If applicable, whether the AE abates upon discontinuation of the test substance

[0274] If applicable, whether the AE re-emerges upon repeated exposure to the test article. SAEs that are not test article related may nevertheless be considered by the participating investigator or medical monitor (or designee) as related to the conduct of the clinical study, i.e., the subject's participation in the study. For example, a protocol-related SAE may be an event that occurs during a washout period or is related to a procedure required by the protocol. The severity of the AE will be assessed according to the National Cancer Institute's (NCI) Common Toxicity Criteria for Adverse Events (CTCAE), version 5.0. For toxicities not defined in the NCI CTCAE, the following definitions will be used: -Mild (Grade 1): The AE is noticed by the subject but does not interfere with daily activities. The AE does not require discontinuing administration of the test article or reducing the dose. - Moderate (Grade 2): The AE interferes with daily activities but responds to symptomatic treatment or rest. The AE may require a reduction in the dose of the test article but does not require discontinuation of administration. - Severe (Grade 3): The AE significantly limits the subject's ability to perform daily activities despite symptomatic treatment. Furthermore, the AE leads to discontinuation or dose reduction of the test article. - Life-threatening (Grade 4): The AE requires that administration of the test article be discontinued. The subject is at immediate risk of death.

[0275] 6.2.2.Results A first cohort of 10 subjects was treated.

[0276] [Table 5]

[0277] Results from the first cohort are shown in Figures 8A-8J.

[0278] Compared to standard of care, CER-001 rapidly improved biomarkers of inflammation, leukocytosis, and endothelial dysfunction and prevented patients from declining into acute kidney injury. CER-001 treatment was well tolerated at all dose levels (5, 10, and 20 mg / kg twice daily). No treatment-related serious side effects were observed in this critically ill patient population.

[0279] VCAM and ICAM The study continued with a total of 20 subjects, n=5 per treatment group. Figures 9A-9F show the change in VCAM in the standard care and CER-001 groups as measured by ELISA. Statistical significance was assessed using mixed model ANOVA (ns: p>0.05). Figure 9A: Change in VCAM from baseline in the SOC and aggregate CER-001 groups. Figure 9B: Change in VCAM from baseline in the SOC and respective study groups. Figure 9C: Change in VCAM reported as a percentage of peak VCAM levels (peak=100%) in the SOC and aggregate CER-001 groups. The effect of treatment x study day on peak was p<0.0001. Figure 9D: Change in VCAM in the SOC and aggregate CER-001 groups broken down by whether the subject was enrolled from the ICU or nephrology department of the center. Figure 9E: Change in VCAM from baseline for each subject in the SOC and aggregate CER-001 groups. FIG. 9F: Change in VCAM from baseline for each subject in each group.

[0280] In general, treatment regimens providing CER-001 in addition to SOC reduced VCAM more than SOC alone. The results are summarized in the table below.

[0281] [Table 6]

[0282] Figures 10A-F show the change in ICAM in standard care (SOC) and experimental (CER-001) groups as measured by ELISA. Statistical significance was assessed using mixed model ANOVA (ns: p>0.05). Figure 10A: Change in ICAM from baseline in SOC and aggregate CER-001 groups. Figure 10B: Change in ICAM from baseline in SOC and each study group. Figure 10C: Change in ICAM reported as a percentage of peak ICAM levels (peak=100%) in SOC and aggregate CER-001 groups. The effect of treatment x study day on peak was p<0.0001. Figure 10D: Change in ICAM in SOC and aggregate CER-001 groups broken down by whether the subject was enrolled from the ICU or nephrology department of the center. Figure 10E: Change in ICAM from baseline for each subject in SOC and aggregate CER-001 groups. FIG. 10F: Change in ICAM from baseline for each subject in each study group.

[0283] In general, treatment regimens providing CER-001 in addition to SOC reduced ICAM more than SOC alone. The results are summarized in the table below.

[0284] [Table 7]

[0285] 6.2.2.2.Leukocytes Figures 18A-F show the change in white blood cell counts for the standard of care (SOC) and CER-001 groups. Figure 18A: Change in white blood cell counts from baseline for the SOC and aggregate CER-001 groups. Figure 18E shows the individual data points summarized in Figure 18A. Figure 18B: Change in white blood cell counts from baseline for the SOC and respective CER-001 groups. Figure 18F shows the individual data points summarized in Figure 18B. Figure 18C: Change in white blood cell counts for the SOC and aggregate CER-001 groups reported as a percentage of peak white blood cell counts (peak = 100%). The effect of treatment x study day on peak was p = 0.5492. Figure 18D: Change in white blood cell counts for the SOC and aggregate CER-001 groups broken down by whether subjects were enrolled from the ICU or nephrology department of the center. Units for Figures 18A, 18B, and 18D: cells / microliter.

[0286] [Example 3] 6.3. Example 3: CER-001 Therapy to Treat CRS Secondary to Covid-19 Infection COVID-19 infects host cells through binding of the viral spike protein (SARS-2-S) to the cell surface receptor angiotensin-converting enzyme 2 (ACE2), and HDL scavenger receptor type B 1 (SR-B1) facilitates ACE2-dependent viral entry. (Wei et al., Nature Metabolism doi.org / 10.1038 / s42255-020-00324-0). Without being bound by theory, it is believed that lipid-binding protein-based complexes such as CER-001 may provide therapeutic benefit (e.g., reducing the severity and / or duration of CRS) in subjects with COVID-19 infection through competitive binding to SR-B1, thereby limiting the virus' ability to infect additional cells.

[0287] A pilot study will be conducted to investigate the safety and efficacy of seven infusions of CER-001 in patients with CRS secondary to COVID-19 infection. The study will consist of nine visits: · Pre-dosing (Baseline) Visit: Assessment of baseline inflammatory markers and safety labs. Dosing Visit: Seven doses (doses 1-7) are administered as a once daily infusion over a 7 day period. IL-6 is measured daily from pre-infusion samples. Follow-up visit: Patients will have their final evaluation on day 8. Inflammatory markers and safety labs will be measured.

[0288] The study flow diagram is shown in Figure 6.

[0289] 6.3.1. Selection of study subjects 6.3.1.1. Inclusion criteria Eligible patients who meet the following criteria will be enrolled in the study: 1. Adult males or non-pregnant females ≥ 18 years of age at the time of enrollment. 2. Have laboratory-confirmed novel coronavirus (COVID-19) infection as determined by polymerase chain reaction (PCR), or other commercial or public health assay, in an oropharyngeal or anal specimen within 72 hours prior to hospitalization. 3. Illness of any duration and at least one of the following: a. Radiographic infiltrates on imaging (chest x-ray, CT scan, etc.), or b. Clinical assessment (evidence of rales / moist rales on physical exam) and SpO2 ≤ 93% on air, or c. Requiring artificial respiration and / or supplemental oxygen; or d. Persistent fever in the past 24 hours that is refractory to NSAIDs or steroids 4. Serum IL-6 ≥ 3x upper limit of normal 5. Females of childbearing potential who agree and promise to use acceptable forms of contraception throughout the study. Acceptable forms of contraception for this study are defined as blockade plus hormonal therapy (implants, injections, oral contraceptives, and IUDs) or abstinence.

[0290] 6.3.1.1. Exclusion criteria Patients who meet the following criteria will be excluded from the study: 1. Patients weighing more than 100 kg 2. Alanine transaminase / aspartate transaminase (ALT / AST) > 5x upper limit of normal. 3. Stage 4 severe chronic kidney disease or requiring dialysis (i.e., estimated glomerular filtration rate (eGFR) < 30ml / min / 1.73m^2) 4. Hemoglobin < 80g / L 5. White blood cell count<2.0×10^9 6. Platelet count <50×10^9 7. Pregnancy or breastfeeding. 8. Anticipated transfer to another hospital outside the study site within 72 hours. 9. Expected life span is not to exceed 7 days. 10. Patients have used an investigational drug within 30 days of the first dose of CER-001.

[0291] 6.3.1.2. Limitations during the study There are no patient restrictions other than those outlined in the inclusion / exclusion criteria above.

[0292] 6.3.1.3. Withdrawal criteria Reasons for patient withdrawal from study drug may include, but are not limited to: At the request of investigators, for safety reasons such as serious adverse reactions, For other reasons, such as investigator request or patient non-compliance, - At the request of the patient or for reasons of durability For other reasons, such as at the patient's request or withdrawal of informed consent.

[0293] Discontinuation of study drug alone does not constitute discontinuation or withdrawal from the study. Patients will continue to be followed as if they had completed the treatment phase. Patients who discontinue study drug prematurely (e.g., before completion of the 7th dose) will undergo end of study evaluations whenever possible.

[0294] 6.3.2. Patient Treatment 6.3.2.1. Research Products CER-001 is provided frozen in 20 mL vials containing approximately 18 mL of product at a concentration of 8 mg / mL (ApoA-I content). CER-001 is dosed by weight. All doses are thawed and then diluted to a volume of 250 mL with normal saline.

[0295] Dosing occurs at each of seven dosing visits. At each of these visits, patients receive a single IV infusion of CER 001 (20 mg / kg) over a 24-hour period using an infusion pump. Patients are pretreated with an antihistamine prior to each CER-001 dose (e.g., dexchlorphenylamine 5 mg or hydroxyzine 100 mg) to avoid any potential infusion reactions.

[0296] 6.3.2.2. Interruption or discontinuation of study medication Patients will be discontinued or withdrawn from study medication if any of the following occur: Any drug-related adverse event or other reason that, in the investigator's opinion, compromises the patient's participation in the trial or the interpretation of the trial data (e.g., serious intercurrent illness requiring additional care measures or preventing further medication). Significant durability issues

[0297] Upon discontinuation of study drug, the study site will document the reason for drug discontinuation, continue to follow the patient clinically, and make every attempt to restart study drug within 2 days of study drug discontinuation, unless otherwise contraindicated.

[0298] 6.3.3. Concomitant Treatment All non-experimental treatments were permitted to be administered concomitantly during patient participation in this study. Any medications taken by patients other than the study drug specified by the protocol will be considered concomitant medications and will be recorded in the study records.

[0299] 6.3.4. Prohibited drugs No drugs have been excluded.

[0300] 6.3.5. Monitoring Patient Compliance CER-001 will be administered under direct observation in a hospital setting.

[0301] Efficacy assessment Efficacy evaluation Inflammatory markers include: CRP, D-dimer, ferritin, IL-6, IL-8, GM-CSF, MCP1, and TNF-α.

[0302] Efficacy parameters (a) Primary efficacy parameters The primary efficacy parameter is the change in IL-6 from baseline to day 8. Baseline is defined as the average of measurements taken at the baseline visit and pre-dose on day 1.

[0303] (b) Secondary efficacy parameters Secondary efficacy parameters included change from baseline to day 8 in the inflammatory markers CRP, D-dimer, ferritin, IL-8, GM-CSF, MCP1, and TNF-α.

[0304] Safety Assessment 6.3.7.1 Safety parameters (a) Pregnancy testing (if applicable) Women of childbearing potential will have documentation of a negative pregnancy test taken any time during hospitalization and prior to dosing.

[0305] (b) Clinical Safety Tests Blood samples will be taken for chemical and hematological analyses at two time points: baseline and day 8. The following tests will be performed by the local hospital laboratory:

[0306] [Table 8]

[0307] 6.3.8.Results IL-6 levels are reduced from baseline to day 8. Secondary efficacy parameters are also reduced from baseline to day 8, indicating that CER-001 treatment can be used to treat CRS and reduce serum levels of inflammatory markers.

[0308] [Example 4] 6.4. Example 4: CER-001 Therapy for Treating CRS Secondary to Covid-19 Infection - Additional Treatment Protocols This example is a study of CER-001 treatment in COVID-19 patients with severe cytokine release syndrome and renal injury.

[0309] 6.4.1.Selection of Object 6.4.1.1. Inclusion criteria Eligible patients will meet the following criteria prior to being enrolled in the study: 1. Adult males or non-pregnant females aged 18 years or older at the time of enrollment. 2. Have laboratory-confirmed novel coronavirus infection as determined by polymerase chain reaction (PCR), or other commercial or public health assay, in an oropharyngeal or anal specimen within 72 hours prior to hospitalization. 3. Illness of any duration and at least one of the following: Radiographic infiltrates on imaging (chest x-ray, CT scan, etc.), or Clinical evaluation (evidence of rales / moist rales on physical exam) and SpO2 <93% on air, or Require artificial respiration and / or supplemental oxygen; or Persistent fever in the past 24 hours that is refractory to NSAIDs or steroids 4. Serum IL-6 > 3x upper limit of normal 5. Females of childbearing potential who agree and promise to use acceptable forms of contraception throughout the study. Acceptable forms of contraception for this study are defined as blockade plus hormonal therapy (implants, injections, oral contraceptives, and IUDs) or abstinence.

[0310] 6.4.1.2. Exclusion criteria Patients who meet any of the following criteria will be excluded from the study. 1. Medical history suggesting allergy to CER-001 2. Pregnancy or breastfeeding. 3. Anticipated transfer to another hospital within 72 hours. 4. The expected life span is not to exceed 7 days. 5. Patients have used an investigational drug within 30 days of the first dose of CER-001.

[0311] Treatment 6.4.2.1. Treatment administered Patients will be pretreated with an antihistamine prior to each CER-001 dose (eg, dexchlorphenylamine 5 mg or hydroxyzine 100 mg) to avoid any potential infusion reactions.

[0312] Patients will receive an IV infusion of CER-001 at a dosage of 15 mg / kg BID for three consecutive days. At the investigator's discretion, patients may receive up to two additional doses.

[0313] Patients may be discontinued or withdrawn from the study drug if any of the following occur: 1. Any drug-related adverse event or other reason that, in the investigator's opinion, compromises the patient's participation in the trial or the interpretation of the trial data (e.g., serious intercurrent illness requiring additional care measures or preventing further medication), 2. Significant durability issues.

[0314] Upon discontinuation of study drug, the study site will document the reason for drug discontinuation, continue to follow the patient clinically, and make every attempt to restart study drug within 2 days of study drug discontinuation, unless otherwise contraindicated.

[0315] Reasons for withdrawal from study drug may include, but are not limited to: 1. At the request of the investigator, for safety reasons such as serious adverse reactions 2. For other reasons, such as investigator request or patient noncompliance. 3. At the patient's request or for reasons of durability 4. For other reasons, such as at the patient's request or withdrawal of informed consent.

[0316] Discontinuation of study drug alone does not constitute discontinuation or withdrawal from the study. Patients will continue to be followed as if they had completed the treatment phase. Patients who discontinue study medication prematurely (e.g., before completion of the third dose) will undergo end of study evaluations whenever possible.

[0317] 6.4.2.2. Dose Changes In case of clinical need defined by the principal investigator, the dose of the drug may be reduced or increased.

[0318] Concomitant medications / treatments All non-experimental treatments were permitted to be administered concomitantly during patient participation in this study. Any medications taken by patients other than the study drug specified by the protocol will be considered concomitant medications and will be recorded in the study records.

[0319] Research evaluation The following procedures will be performed during the baseline visit: The following tests will be performed by the local hospital laboratory: 1. Informed consent 2. Medical history includes recording past and present illnesses and collecting the subject's demographic data (date of birth, sex, and race). 3. Physical examination with review of the system, height and weight, BMI, and waist circumference 4. Vital signs (pulse, blood pressure, and oral, ear, axillary, or core temperature). 5. Review of inclusion / exclusion criteria. 6. Adverse events will be recorded starting from the time informed consent is obtained. 7. Previous medications will be collected starting 4 weeks prior to the first dose of test substance. All current medications will be recorded. 8. Complete Blood Count (CBC) - Includes white blood cell (WBC) differential, platelet count, red blood cell (RBC), hemoglobin (Hb), and hematocrit (Hct). 9. Fasting Chemistry Panel / Electrolytes: Includes sodium, potassium, chloride, blood urea nitrogen (BUN, or urea), serum creatinine, calculated clearance creatinine (CKD-EPI), glucose, calcium, phosphorus, total protein, uric acid, AST, ALT, □GT, ALP, total and direct bilirubin, albumin, total cholesterol, HDL, LDL, triglycerides, LDH, CPK, 10. ABG (to assess respiratory and / or metabolic disorders) 11. ApoA-I (for pharmacokinetic and pharmacodynamic evaluation) 12. Coagulation tests-include prothrombin time (PT) (expressed as international normalized ratio [INR]) and partial thromboplastin time (PTT). 13. Urinalysis- Includes specific gravity, pH, protein / albumin assessment, glucose, ketones, and hemoglobin / blood. 14. Microalbuminuria and proteinuria, g / 24 hours 15. Serum or urine pregnancy test (for females of childbearing potential) within 7 days prior to randomization. 16. Pharmacokinetic and pharmacodynamic assessments will include apoA-I and total cholesterol levels. 17. Inflammatory markers include CRP, PCT, D-dimer, ferritin, IL-6, IL-8, GM-CSF, MCP1, and TNF-α.

[0320] As reported in FIG. 7, clinical and laboratory parameters were monitored from baseline to the final visit on day 8, which included the following procedures: 1. Recording of adverse events and concomitant medications 2. Review of pertinent laboratory information 3. Physical Examination 4. Assess vital signs (pulse, blood pressure, and oral, ear, axillary, or core temperature). 5. Continuously record adverse events and concomitant medications 6. Complete Blood Count (CBC) - Includes white blood cell (WBC) differential, platelet count, red blood cell (RBC), hemoglobin (Hb), and hematocrit (Hct). 7. Fasting Chemistry Panel / Electrolytes: Includes sodium, potassium, chloride, blood urea nitrogen (BUN, or urea), serum creatinine, calculated clearance creatinine (CKD-EPI), glucose, calcium, phosphorus, total protein, uric acid, AST, ALT, □GT, ALP, total and direct bilirubin, albumin, total cholesterol, HDL, LDL, triglycerides, LDH, CPK 8. ABG (to assess respiratory and / or metabolic disorders) 9. ApoA-I (for pharmacokinetic and pharmacodynamic evaluation) 10. Coagulation tests-include prothrombin time (PT) (expressed as international normalized ratio [INR]) and partial thromboplastin time (PTT). 11. Urinalysis-includes specific gravity, pH, protein / albumin assessment, glucose, ketones, and hemoglobin / blood. 12. Microalbuminuria and proteinuria, g / 24 hours 13. Inflammatory markers include CRP, PCT, D-dimer, ferritin, IL-6, IL-8, GM-CSF, MCP1, and TNF-α.

[0321] 6.4.4. Reporting of Adverse Events (AEs) An AE is any untoward medical occurrence associated with the use of an investigational product (active or placebo drug, biological agent, or device) in a clinical investigational patient, and does not necessarily have a causal relationship to the product. Thus, an AE can be any undesirable and unintended sign (e.g., an abnormal laboratory finding), symptom, or disease that is temporally related to the use of the investigational product, whether or not it is considered related to the investigational product.

[0322] Adverse events may include: Symptoms described by the patient Clinically significant changes in the patient's physical examination or other signs observed by the investigator or medical staff Laboratory abnormalities (clinical tests) that reflect a change from baseline and / or that may result in a change in administration of the investigational product or a change in medical care (diagnostic or therapeutic). Conditions present at baseline that either worsened or resolved and then recurred.

[0323] Patients are assessed for the status of new and pre-existing AEs at each study visit.

[0324] 6.4.5.Results IL-6 levels and other inflammatory markers decrease from baseline to day 8.

[0325] [Example 5] 6.5. Example 5: CER-001 Therapy to Treat Ischemia / Reperfusion AKI This example is a study of CER-001 therapy for treating ischemia / reperfusion AKI.

[0326] Materials and Methods Pigs, weighing 45-60 kg, are fasted for 24 h prior to the study. All animals are administered azaperone (8 mg kg 2 -1 ) and atropine (0.03 mg kg -1The animals are premedicated with an intramuscular mixture of 1-(2-aminopropyl)-2-propanediol (1-NaCl) and 1-(2-methylpropyl)-2-propanediol (1-NaCl). After anesthesia, both kidneys are approached through a midline abdominal incision. The renal artery and vein are then isolated, and a vascular loop is placed around the renal artery using right-angled forceps. Warm ischemia is induced for 60 min by pulling the vascular loop. Ischemia is followed by 3 h of reperfusion, with half of the animals receiving CER-001 administered directly through the renal artery for 5 min before the start of reperfusion. At 24 h, the animals are euthanized by IV administration of 1-mL / kg BW pentobarbital. The kidneys are then collected for analysis.

[0327] 6.5.2.Results CER-001 attenuates ischemia / reperfusion AKI.

[0328] [Example 6] 6.6. Example 6: Lipid-binding protein molecule therapy in a model of LPS-induced vascular endothelial injury The ability of the ApoA-I-containing complex CER-001 to attenuate sepsis-induced injury to the vascular endothelium was evaluated in a lipopolysaccharide (LPS)-induced in vitro model.

[0329] Materials and Methods 6.6.1.1.Cell culture Human umbilical vein endothelial cells (HUVECs, ECs) were purchased from the American Type Culture Collection (ATCC-LGC Standards Srl, Sesto San Giovanni, Milan, Italy). ECs were maintained in their recommended medium, EndGro (Merck Millipore, Darmstadt, Germany).

[0330] Peripheral blood mononuclear cells (PBMCs) were isolated by gradient centrifugation from buffy coats of healthy donors (selected from a research repository) using the Ficoll-Hypaque method as previously described (Sallustio, et al., 2021, Nephrol Dial Transplant 36, 452-464). PBMCs were maintained in their recommended medium (ibid.).

[0331] When cells became confluent, they were stimulated with LPS 0.3 μg / ml, 4 μg / ml (E. Coli O111:B4, Sigma-Aldrich, Milan, Italy) and CER-001 50, 100, and 500 μg / ml for the indicated periods.

[0332] PBMC culture supernatants were collected and analyzed for TNF-α by ELISA (R&D Systems, Minneapolis, MN, USA).

[0333] 6.6.1.2. Cell proliferation assay ECs and PBMCs were incubated with 0.3 μg / ml LPS and / or 50 and 500 μg / ml CER-001 for 60 min and 24 h. Proliferation rates were measured by MTT cell proliferation assay kit according to the manufacturer's instructions (Sigma Aldrich). Briefly, 3 × 10 4 Cells / well were seeded in 96-well plates, and then cells were treated with LPS and CER-001 as indicated, after which the absorbance at 570 nm was measured spectrophotometrically.

[0334] 6.6.1.3. Immunophenotypic analysis After stimulation, ECs were permeabilized with an IntraPrep kit (Instrumentation Laboratory) and incubated with unconjugated primary antibody p-ENOS (Abcam) for 25 min at 4° C. Then, cells were washed and labeled with secondary antibody AlexaFluor 488 (Molecular Probes) for 25 min at 4° C. Finally, cells were washed twice and resuspended in FACS buffer for acquisition.

[0335] PBMCs were stained with the following monoclonal antibody, CD14 Monoclonal Antibody (61D3)-PE, (eBioscience™, Thermo Fisher Scientific, Italy) for 20 minutes at room temperature in the dark, washed twice and resuspended in FACS buffer, after which the stained PBMCs were acquired.

[0336] Data were acquired using a FC500 (Beckman Coulter) flow cytometer and analyzed using Kaluza software. Three independent studies were performed on both EC and PBMC. Positive areas were determined using isotype-matched mAbs, and a total of 104 events were acquired for each sample.

[0337] 6.6.1.4.Statistical analysis Data are representative of three independent studies. Data are presented as mean ± standard deviation (SD) and compared using Student's t test.

[0338] 6.6.2.Results The effects of LPS and CER-001 on endothelial cells and endothelial nitric oxide synthase (eNOS) activation were analyzed. The results of the MTT cell viability assay in Figure 11 showed a slight decrease in proliferation after LPS stimulation. CER-001 at 50 and 500 μg / ml did not affect endothelial viability. Endothelial cells treated with LPS and CER-001, both at 50 and 500 μg / ml, increased the proliferation rate compared to cells stimulated with LPS, especially at the highest concentration.

[0339] The production of eNOS has been described as a marker of vascular endothelial integrity (Zhao, et al., 2015, J Pharmacol Sci 129, 83-94). In our in vitro model, eNOS phosphorylation and activation (Figures 12-13) were altered by LPS and upregulated by CER-001. Specifically, a strong reduction in eNOS (phospho-S1177) (p-ENOS) was observed after 60 min of LPS stimulation compared to basal and VEGF (positive control). Supplementation with 500 μg / ml CER-001 completely reversed the LPS effect. (In Figure 13, representative data from one of three total studies are shown. Histograms show p-ENOS expression levels).

[0340] Furthermore, CER-001 modulated the response of peripheral blood mononuclear cells (PBMCs) stimulated for 24 hours with 0.3 μg / ml LPS and / or 50 and 500 μg / ml CER-001, decreasing mCD14 expression and TNF-α secretion. As shown in FIG. 14, MTT assays showed no significant differences in cell viability versus basal for the above conditions. PBMC culture supernatants were analyzed by ELISA and the results are shown in FIG. 15. 24 hours after LPS stimulation, PBMCs increased the synthesis of TNF-α. Stimulation of PBMCs with 50 and 500 μg / m CER-001 alone did not affect TNF-α production. Addition of both 50 and 500 μg / m CER-001 into the culture medium of LPS-activated PBMCs reversed the LPS effect. Alternatively, FACS demonstrated a strong upregulation of CD14 surface expression by PCMB 24 hours after LPS stimulation (Figure 16). PBMCs treated with a combination of LPS and CER-001 maintained CD14 expression at basal levels (Figure 17).

[0341] 7. Incorporation by Reference All publications, patents, patent applications, and other documents cited in this application are incorporated by reference herein in their entirety for all purposes to the same extent as if each individual publication, patent, patent application, or other document was individually indicated to be incorporated by reference for all purposes.

[0342] Any discussion of documents, acts, materials, devices, articles or the like which has been included in this specification is solely for the purpose of providing the context of the present disclosure and is not to be taken as an admission that any or all of these matters form part of the prior art base or were common general knowledge in the field relevant to the present disclosure, as they existed anywhere prior to the priority date of this application.

[0343] 8. Specific embodiments Various aspects of the disclosure are described in the embodiments set forth in the numbered paragraphs of Group 1 below.

[0344] Group 1: 1. A method of treating a subject having or at risk of developing leukocytosis, comprising administering to the subject a lipid-binding protein-based conjugate in a dose effective to reduce the subject's white blood cell count. 2. The method of embodiment 1, wherein the subject has one or more symptoms associated with leukocytosis. 3. A method of treating a subject having one or more symptoms associated with leukocytosis, comprising administering to the subject a lipid-binding protein-based complex in a dose effective to reduce the subject's white blood cell count and / or ameliorate one or more of the one or more symptoms associated with leukocytosis. 4. The method of embodiment 2 or embodiment 3, wherein the one or more symptoms include fever. 5. The method of any one of embodiments 2 to 4, wherein the one or more symptoms include bleeding or bruising. 6. The method of any one of embodiments 2 to 5, wherein the one or more symptoms include sweating. 7. The method of any one of embodiments 2 to 6, wherein the one or more symptoms include pain or tingling in the legs, arms, or abdomen. 8. The method of any one of embodiments 2 to 7, wherein the one or more symptoms include one or more vision problems. 9. The method of embodiment 8, wherein the one or more vision problems include blurred vision. 10. The method of embodiment 8 or embodiment 9, wherein the one or more vision problems include diplopia. 11. The method of any one of embodiments 8 to 10, wherein the one or more vision problems include blind spots. 12. The method of any one of embodiments 8 to 11, wherein the one or more vision problems include cloudy vision. 13. The method of any one of embodiments 8 to 12, wherein the one or more vision problems include dim vision. 14. The method of any one of embodiments 2 to 13, wherein one or more symptoms include slurred thinking. 15. The method of any one of embodiments 2 to 14, wherein the one or more symptoms include loss of appetite. 16. The method of any one of embodiments 2 to 15, wherein the one or more symptoms include dyspnea (e.g., shortness of breath, below-normal blood oxygen levels). 17. The method of any one of embodiments 2 to 16, wherein the dose of the lipid-binding protein-based complex is effective to ameliorate one or more of the one or more symptoms associated with leukocytosis. 18. The method of any one of embodiments 1 to 17, wherein the subject has or is at risk of developing leukocytosis due to inflammation, infection, white blood cell disorder, physical stress, emotional stress, medication, or an allergic reaction. 19. The method of embodiment 18, wherein the subject has or is at risk of developing leukocytosis due to inflammation. 20. The method of embodiment 18, wherein the subject has or is at risk of developing leukocytosis due to an infection. 21. The method of embodiment 18, wherein the subject has or is at risk of developing leukocytosis due to a white blood cell disorder. 22. The method of embodiment 18, wherein the subject has or is at risk of developing leukocytosis due to physical stress, and optionally the physical stress results from an epileptic seizure, anesthesia, or excessive exertion. 23. The method of embodiment 18, wherein the subject has or is at risk of developing leukocytosis due to emotional stress. 24. The method of embodiment 18, wherein the subject has or is at risk of developing drug-induced leukocytosis, and optionally the drug is a steroid, a corticosteroid, lithium, or a beta-agonist. 25. The method of embodiment 18, wherein the subject has or is at risk of developing leukocytosis due to an allergic reaction. 26. The method of any one of embodiments 1 to 25, wherein the subject has an infection. 27. The method of embodiment 26, wherein the infectious disease is a viral infection, optionally wherein the viral infection is a coronavirus infection, optionally wherein the coronavirus infection is COVID-19. 28. The method of embodiment 26, wherein the infection is a bacterial infection. 29. The method of embodiment 26, wherein the infection is a fungal infection. 30. The method of embodiment 26, wherein the infectious disease is a parasitic infection. 31. The method of any one of embodiments 1 to 30, wherein the subject has diabetes. 32. The method of any one of embodiments 1 to 31, wherein the subject has a white blood cell disorder. 33. The method of embodiment 32, wherein the leukocyte disorder is a primary bone marrow disorder (e.g., acute leukemia, chronic leukemia, or a myeloproliferative disorder). 34. The method of embodiment 32, wherein the leukocyte disorder is neutrophilia, and optionally, the neutrophilia is idiopathic neutrophilia, optionally chronic idiopathic neutrophilia. 35. The method of embodiment 32, wherein the white blood cell disorder is hemolytic anemia. 36. The method of embodiment 32, wherein the white blood cell disorder is thrombocytopenia, and optionally, the thrombocytopenia is idiopathic thrombocytopenia or essential thrombocytopenia. 37. The method of embodiment 32, wherein the white blood cell disorder is cancer, optionally bone cancer or blood cancer. 38. The method of embodiment 32, wherein the leukocyte disorder is a myeloproliferative disorder or a blood cancer. 39. The method of embodiment 38, wherein the leukocyte disorder is a myeloproliferative disorder. 40. The method of embodiment 38, wherein the white blood cell disorder is a blood cancer. 41. The method of embodiment 38, wherein the leukocyte disorder is lymphoma. 42. The method of embodiment 38, wherein the white blood cell disorder is leukemia. 43. The method of embodiment 42, wherein the leukemia is chronic leukemia. 44. The method of embodiment 42, wherein the leukemia is acute leukemia. 45. The method of embodiment 42, wherein the leukemia is lymphocytic leukemia, chronic eosinophilic leukemia, chronic myelogenous leukemia, or chronic neutrophilic leukemia. 46. ​​The method of embodiment 38, wherein the white blood cell disorder is polycythemia vera. 47. The method of embodiment 38, wherein the leukocyte disorder is myelofibrosis, and optionally, the myelofibrosis is primary myelofibrosis. 48. The method of any one of embodiments 1 to 47, wherein the subject has leukocytosis. 49. Target is 11×10 9 49. The method of embodiment 48, wherein the patient has a white blood cell count of more than cells / L. 50. The target is 12×10 9 The method of embodiment 48 or embodiment 49, wherein the patient has a white blood cell count of more than 10 cells / L. 51. Target is 15×10 9 The method of any one of embodiments 48 to 50, wherein the patient has a white blood cell count of more than cells / L. 52. The target is 20×10 9The method of any one of embodiments 48 to 51, wherein the patient has a white blood cell count of more than cells / L. 53. The target is 100 x 10 9 The method of embodiment 48 or embodiment 49, wherein the patient has a white blood cell count of less than 10 cells / L. 54. The target is 100 x 10 9 The method of any one of embodiments 48 to 52, wherein the patient has a white blood cell count of less than 1 / L. 55. Target is 50×10 9 The method of any one of embodiments 48 to 54, wherein the patient has a white blood cell count of less than 100 / L. 56. Target is 25×10 9 The method of any one of embodiments 48 to 55, wherein the patient has a white blood cell count of less than 100 / L. 57. Target is 15×10 9 The method of any one of embodiments 48 to 56, wherein the patient has a white blood cell count of less than 10 cells / L. 58. Target is 11×10 9 pcs / L~50×10 9 The method of any one of embodiments 48 to 57, wherein the patient has a white blood cell count of 100 / L. 59. Target is 11×10 9 pcs / L~25×10 9 The method of any one of embodiments 48 to 57, wherein the patient has a white blood cell count of 100 / L. 60. Target is 25×10 9 pcs / L~50×10 9 The method of any one of embodiments 48 to 57, wherein the patient has a white blood cell count of 100 / L. 61. Target is 50×10 9 pcs / L~100×10 9 The method of any one of embodiments 48 to 51, wherein the patient has a white blood cell count of 100 / L. 62. Target is 11×10 9 pcs / L~15×10 9 The method of any one of embodiments 48 to 57, wherein the patient has a white blood cell count of 100 / L. 63. Target is 15×10 9 pcs / L~20×10 9The method of any one of embodiments 48 to 57, wherein the patient has a white blood cell count of 100 / L. 64. A method according to any one of embodiments 1 to 47, wherein the subject is at risk for leukocytosis. 65. A method of treating a subject having endothelial dysfunction, comprising administering to the subject a dose of a lipid-binding protein-based complex, wherein the dose is a high dose. 66. A method of treating a subject experiencing or who has experienced an acute coronary syndrome or stroke, comprising administering to the subject a dose of a lipid-binding protein-based complex, wherein the dose is a high dose. 67. The method of embodiment 65 or embodiment 66, wherein the subject is experiencing an acute coronary syndrome, optionally wherein the acute coronary syndrome is myocardial infarction (e.g., ST-elevation myocardial infarction or non-ST-elevation myocardial infarction) or unstable angina. 68. The method of embodiment 65 or embodiment 66, wherein the subject has experienced an acute coronary syndrome, and optionally the acute coronary syndrome is myocardial infarction (e.g., ST-segment elevation myocardial infarction or non-ST-segment elevation myocardial infarction) or unstable angina. 69. The method of embodiment 65 or embodiment 66, wherein the subject has experienced a stroke. 70. The method of embodiment 65 or embodiment 66, wherein the subject has experienced a stroke. 71. A method of treating a subject having or at risk of developing carditis, comprising administering to the subject a dose of a lipid-binding protein-based complex, optionally wherein the carditis is myocarditis and / or pericarditis. 72. The method of embodiment 71, wherein the carditis comprises myocarditis. 73. The method of embodiment 71 or embodiment 72, wherein the carditis comprises pericarditis. 74. The method of any one of embodiments 71 to 73, wherein the subject is male. 75. The method of any one of embodiments 71 to 74, wherein the subject is under 60 years of age. 76. The method of any one of embodiments 71 to 74, wherein the subject is under 50 years of age. 77. The method of any one of embodiments 71 to 74, wherein the subject is under 40 years of age. 78. The method of any one of embodiments 71 to 74, wherein the subject is under 30 years of age. 79. The method of any one of embodiments 71 to 74, wherein the subject is under 20 years of age. 80. The method of any one of embodiments 71 to 79, wherein the subject is at least 5 years old. 81. The method of any one of embodiments 71 to 79, wherein the subject is at least 8 years old. 82. The method of any one of embodiments 71 to 79, wherein the subject is at least 10 years old. 83. The method of any one of embodiments 71 to 79, wherein the subject is at least 12 years old. 84. The method of any one of embodiments 71 to 79, wherein the subject is at least 15 years of age. 85. The method of any one of embodiments 71 to 79, wherein the subject is at least 18 years of age. 86. The method of any one of embodiments 71 to 85, wherein the subject has an infection. 87. The method of embodiment 74, wherein the infectious disease is a viral infection, which is optionally a coronavirus infection, which is optionally COVID-19. 88. The method of embodiment 74, wherein the infection is a bacterial infection. 89. A method according to any one of embodiments 71 to 88, wherein the lipid-binding protein-based conjugate is administered prior to administering the vaccine to the subject. 90. A method according to any one of embodiments 71 to 88, wherein the lipid-binding protein-based complex is administered to the subject concomitantly with the vaccine. 91. A method according to any one of embodiments 71 to 88, wherein the lipid-binding protein-based conjugate is administered after administration of the vaccine to the subject. 92. The method of any one of embodiments 89 to 91, wherein the vaccine is a coronavirus vaccine, optionally a COVID-19 vaccine. 93. The method of any one of embodiments 71 to 92, wherein the subject has carditis. 94. The method of embodiment 93, wherein the dose of the lipid-binding protein-based complex is effective to ameliorate one or more symptoms of carditis. 95. The method of any one of embodiments 71 to 92, wherein the subject is at risk for carditis. 96. The method of embodiment 95, wherein the dose of the lipid-binding protein-based complex is effective to prevent carditis or reduce the severity of carditis. 97. The method of any one of embodiments 1 to 96, wherein the dose is effective to reduce circulating VCAM-1 and / or ICAM-1 in the subject. 98. The method of embodiment 97, wherein the dose is effective to reduce the subject's circulating VCAM-1 by at least 100 ng / mL within 2 days of the first administration of the lipid-binding protein-based conjugate. 99. The method of embodiment 97, wherein the dose is effective to reduce the subject's circulating VCAM-1 by at least 200 ng / mL within 2 days of the first administration of the lipid-binding protein-based conjugate. 100. The method of embodiment 97, wherein the dose is effective to reduce circulating VCAM-1 in the subject by at least 300 ng / mL within 2 days of the first administration of the lipid binding protein-based conjugate. 101. The method of any one of embodiments 97 to 100, wherein the dose is effective to reduce the subject's circulating VCAM-1 to 400 ng / mL within 2 days of the first administration of the lipid-binding protein-based conjugate. 102. The method of any one of embodiments 97 to 101, wherein the dose is effective to reduce the subject's circulating VCAM-1 by at least 100 ng / mL within 5 days of the first administration of the lipid-binding protein-based conjugate. 103. The method of any one of embodiments 97 to 101, wherein the dose is effective to reduce the subject's circulating VCAM-1 by at least 200 ng / mL within 5 days of the first administration of the lipid-binding protein-based conjugate. 104. The method of any one of embodiments 97 to 101, wherein the dose is effective to reduce the subject's circulating VCAM-1 by at least 300 ng / mL within 5 days of the first administration of the lipid-binding protein-based conjugate. 105. The method of any one of embodiments 97 to 104, wherein the dose is effective to reduce the subject's circulating VCAM-1 to 400 ng / mL within 5 days of the first administration of the lipid-binding protein-based conjugate. 106. The method of any one of embodiments 97 to 105, wherein the dose is effective to reduce the subject's circulating ICAM-1 by at least 50 ng / mL within 2 days of the first administration of the lipid binding protein-based complex. 107. The method of any one of embodiments 97 to 105, wherein the dose is effective to reduce the subject's circulating ICAM-1 by at least 75 ng / mL within 2 days of the first administration of the lipid binding protein-based complex. 108. The method of any one of embodiments 97 to 105, wherein the dose is effective to reduce the subject's circulating ICAM-1 by at least 100 ng / mL within 2 days of the first administration of the lipid binding protein-based complex. 109. The method of any one of embodiments 97 to 108, wherein the dose is effective to reduce the subject's circulating ICAM-1 to 125 ng / mL within 2 days of the first administration of the lipid binding protein-based complex. 110. The method of any one of embodiments 97 to 110, wherein the dose is effective to reduce the subject's circulating ICAM-1 by at least 50 ng / mL within 5 days of the first administration of the lipid binding protein-based complex. 111. The method of any one of embodiments 97 to 110, wherein the dose is effective to reduce the subject's circulating ICAM-1 by at least 75 ng / mL within 5 days of the first administration of the lipid-binding protein-based complex. 112. The method of any one of embodiments 97 to 110, wherein the dose is effective to reduce the subject's circulating ICAM-1 by at least 100 ng / mL within 5 days of the first administration of the lipid binding protein-based complex. 113. The method of any one of embodiments 97 to 112, wherein the dose is effective to reduce the subject's circulating ICAM-1 to 125 ng / mL within 5 days of the first administration of the lipid binding protein-based complex. 114. A method according to any one of embodiments 1 to 113, wherein the dose is a high dose. 115. The method of embodiment 114, wherein the high dose is administered for a period of 2 days to approximately 2 weeks, optionally, the high dose is administered for a period of 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, or 15 days. 116. The method of embodiment 115, wherein the high dose is administered over a period of two days. 117. The method of embodiment 115, wherein the high dose is administered over a period of three days. 118. The method of embodiment 115, wherein the high dose is administered over a period of 4 days. 119. The method of embodiment 115, wherein the high dose is administered over a period of 5 days. 120. The method of embodiment 115, wherein the high dose is administered over a period of 6 days. 121. The method of embodiment 115, wherein the high dose is administered over a period of 7 days. 122. The method of embodiment 115, wherein the high dose is administered over a period of 8 days. 123. The method of embodiment 115, wherein the high dose is administered over a period of 9 days. 124. The method of embodiment 115, wherein the high dose is administered over a period of 10 days. 125. The method of embodiment 115, wherein the high dose is administered over a period of 11 days. 126. The method of embodiment 115, wherein the high dose is administered over a period of 12 days. 127. The method of embodiment 115, wherein the high dose is administered over a period of 13 days. 128. The method of embodiment 115, wherein the high dose is administered over a period of 14 days. 129. The method of embodiment 115, wherein the high dose is administered for a period of 15 days. 130. The method of any one of embodiments 114 to 129, wherein the high dose is a collection of 2 to 20 individual doses, optionally wherein the high dose is a collection of 2, 3, 4, 5, 6, 7, 8, 9, or 10 individual doses. 131. The method of embodiment 130, wherein the high dose comprises 2 to 16 individual doses. 132. The method of embodiment 130, wherein the high dose comprises 2 to 12 individual doses. 133. The method of embodiment 130, wherein the high dose comprises 2 to 10 individual doses. 134. The method of embodiment 130, wherein the high dose comprises 2 to 8 individual doses. 135. The method of embodiment 130, wherein the high dose comprises 2 to 6 individual doses. 136. The method of embodiment 130, wherein the high dose comprises 2 to 4 individual doses. 137. The method of embodiment 130, wherein the high dose comprises 4 to 20 individual doses. 138. The method of embodiment 130, wherein the high dose comprises 4 to 16 individual doses. 139. The method of embodiment 130, wherein the high dose comprises 4 to 10 individual doses. 140. The method of embodiment 130, wherein the high dose comprises 4 to 6 individual doses. 141. The method of embodiment 130, wherein the high dose comprises 6 to 20 individual doses. 142. The method of embodiment 130, wherein the high dose comprises 6 to 16 individual doses. 143. The method of embodiment 130, wherein the high dose comprises 6 to 10 individual doses. 144. The method of embodiment 130, wherein the high dose comprises 6 to 8 individual doses. 145. The method of embodiment 130, wherein the high dose comprises 8 to 20 individual doses. 146. The method of embodiment 130, wherein the high dose comprises 8 to 16 individual doses. 147. The method of embodiment 130, wherein the high dose comprises 8 to 10 individual doses. 148. The method of embodiment 130, wherein the high dose comprises 10 to 20 individual doses. 149. The method of embodiment 130, wherein the high dose comprises 10 to 16 individual doses. 150. The method of embodiment 130, wherein the high dose comprises 16 to 20 individual doses. 151. The method of embodiment 130, wherein the high dose comprises two individual doses. 152. The method of embodiment 130, wherein the high dose comprises three individual doses. 153. The method of embodiment 130, wherein the high dose comprises four individual doses. 154. The method of embodiment 130, wherein the high dose comprises five individual doses. 155. The method of embodiment 130, wherein the high dose comprises six individual doses. 156. The method of embodiment 130, wherein the high dose comprises seven individual doses. 157. The method of embodiment 130, wherein the high dose comprises eight individual doses. 158. The method of embodiment 130, wherein the high dose comprises nine individual doses. 159. The method of embodiment 130, wherein the high dose comprises 10 individual doses. 160. The method of any one of embodiments 130 to 159, wherein multiple individual doses are administered daily or twice daily. 161. The method of any one of embodiments 130 to 159, wherein multiple individual doses are administered approximately 12 hours apart. 162. The method of any one of embodiments 130 to 161, in which multiple individual doses are administered at intervals of 2 to 3 days. 163. The method of any one of embodiments 130 to 162, wherein each individual dose is effective to increase HDL levels in the subject. 164. The method of embodiment 163, wherein each individual dose is effective to increase the subject's HDL levels by at least 25%, at least 30%, or at least 35% 2 to 4 hours after administration. 165. The method of embodiment 164, wherein each individual dose is effective to increase the subject's HDL levels by at least 25%, at least 30%, or at least 35% 2 hours after administration. 166. The method of embodiment 164, wherein each individual dose is effective to increase the subject's HDL levels by at least 25%, at least 30%, or at least 35% 3 hours after administration. 167. The method of embodiment 164, wherein each individual dose is effective to increase the subject's HDL levels by at least 25%, at least 30%, or at least 35% 4 hours after administration. 168. The method of any one of embodiments 130 to 167, wherein each individual dose is effective to increase the subject's ApoA-I levels. 169. The method of embodiment 168, wherein each individual dose is effective to increase the subject's ApoA-I levels by at least 25%, at least 30%, or at least 35% 2 to 4 hours after administration. 170. The method of embodiment 169, wherein each individual dose is effective to increase the subject's ApoA-I levels by at least 25%, at least 30%, or at least 35% 2 hours after administration. 171. The method of embodiment 169, wherein each individual dose is effective to increase the subject's ApoA-I levels by at least 25%, at least 30%, or at least 35% 3 hours after administration. 172. The method of embodiment 169, wherein each individual dose is effective to increase the subject's ApoA-I levels by at least 25%, at least 30%, or at least 35% 4 hours after administration. 173. A method according to any one of embodiments 1 to 172, wherein the dose is effective to improve vascular endothelial function in the subject, and optionally, the vascular endothelial function is measured by circulating VCAM-1 and / or ICAM-1. 174. The method of embodiment 173, wherein the dose is effective to reduce the subject's circulating VCAM-1 by at least 100 ng / mL within 2 days of the first administration of the lipid-binding protein-based conjugate. 175. The method of embodiment 173, wherein the dose is effective to reduce the subject's circulating VCAM-1 by at least 200 ng / mL within 2 days of the first administration of the lipid-binding protein-based conjugate. 176. The method of embodiment 173, wherein the dose is effective to reduce the subject's circulating VCAM-1 by at least 300 ng / mL within 2 days of the first administration of the lipid-binding protein-based conjugate. 177. The method of any one of embodiments 173 to 176, wherein the dose is effective to reduce the subject's circulating VCAM-1 to 400 ng / mL within 2 days of the first administration of the lipid binding protein-based conjugate. 178. The method of embodiment 173 to 177, wherein the dose is effective to reduce circulating VCAM-1 in the subject by at least 100 ng / mL within 5 days of the first administration of the lipid-binding protein-based conjugate. 179. The method of embodiment 173 to 177, wherein the dose is effective to reduce circulating VCAM-1 in the subject by at least 200 ng / mL within 5 days of the first administration of the lipid-binding protein-based conjugate. 180. The method of embodiment 173 to 177, wherein the dose is effective to reduce circulating VCAM-1 in the subject by at least 300 ng / mL within 5 days of the first administration of the lipid-binding protein-based conjugate. 181. The method of any one of embodiments 173 to 180, wherein the dose is effective to reduce the subject's circulating VCAM-1 to 400 ng / mL within 5 days of the first administration of the lipid-binding protein-based conjugate. 182. The method of any one of embodiments 173 to 181, wherein the dose is effective to reduce the subject's circulating ICAM-1 by at least 50 ng / mL within 2 days of the first administration of the lipid binding protein-based conjugate. 183. The method of any one of embodiments 173 to 181, wherein the dose is effective to reduce the subject's circulating ICAM-1 by at least 75 ng / mL within 2 days of the first administration of the lipid binding protein-based conjugate. 184. The method of any one of embodiments 173 to 181, wherein the dose is effective to reduce the subject's circulating ICAM-1 by at least 100 ng / mL within 2 days of the first administration of the lipid binding protein-based conjugate. 185. The method of any one of embodiments 173 to 184, wherein the dose is effective to reduce the subject's circulating ICAM-1 to 125 ng / mL within 2 days of the first administration of the lipid binding protein-based conjugate. 186. The method of any one of embodiments 173 to 185, wherein the dose is effective to reduce the subject's circulating ICAM-1 by at least 50 ng / mL within 5 days of the first administration of the lipid binding protein-based conjugate. 187. The method of any one of embodiments 173 to 185, wherein the dose is effective to reduce the subject's circulating ICAM-1 by at least 75 ng / mL within 5 days of the first administration of the lipid binding protein-based conjugate. 188. The method of any one of embodiments 173 to 185, wherein the dose is effective to reduce the subject's circulating ICAM-1 by at least 100 ng / mL within 5 days of the first administration of the lipid-binding protein-based conjugate. 189. The method of any one of embodiments 173 to 188, wherein the dose is effective to reduce the subject's circulating ICAM-1 to 125 ng / mL within 5 days of the first administration of the lipid-binding protein-based conjugate. 190. The method of any one of embodiments 1 to 189, wherein the dose is effective to reduce the subject's white blood cell count by at least 2000 WBC / microliter within two days of the first administration of the lipid-binding protein-based conjugate. 191. The method of any one of embodiments 1 to 189, wherein the dose is effective to reduce the subject's white blood cell count by at least 3,000 WBC / microliter within two days of the first administration of the lipid-binding protein-based conjugate. 192. The method of any one of embodiments 1 to 189, wherein the dose is effective to reduce the subject's white blood cell count by at least 4000 WBC / microliter within two days of the first administration of the lipid-binding protein-based conjugate. 193. The method of any one of embodiments 1 to 189, wherein the dose is effective to reduce the subject's white blood cell count by at least 5,000 WBC / microliter within two days of the first administration of the lipid-binding protein-based conjugate. 194. The method of any one of embodiments 1 to 189, wherein the dose is effective to reduce the subject's white blood cell count by at least 6,000 WBC / microliter within two days of the first administration of the lipid-binding protein-based conjugate. 195. The method of any one of embodiments 1 to 194, wherein the dose is effective to reduce the subject's white blood cell count to 8,000 WBC / microliter within two days of the first administration of the lipid-binding protein-based conjugate. 196. The method of any one of embodiments 1 to 189, wherein the dose is effective to reduce the subject's white blood cell count by at least 2000 WBC / microliter within 5 days of the first administration of the lipid-binding protein-based conjugate. 197. The method of any one of embodiments 1 to 189, wherein the dose is effective to reduce the subject's white blood cell count by at least 3,000 WBC / microliter within 5 days of the first administration of the lipid-binding protein-based conjugate. 198. The method of any one of embodiments 1 to 189, wherein the dose is effective to reduce the subject's white blood cell count by at least 4000 WBC / microliter within 5 days of the first administration of the lipid-binding protein-based conjugate. 199. The method of any one of embodiments 1 to 189, wherein the dose is effective to reduce the subject's white blood cell count by at least 5,000 WBC / microliter within 5 days of the first administration of the lipid-binding protein-based conjugate. 200. The method of any one of embodiments 196 to 199, wherein the dose is effective to reduce the subject's white blood cell count to 6,000 WBC / microliter within two days of the first administration of the lipid-binding protein-based conjugate. 201. The method of any one of embodiments 1 to 200, wherein the dose is effective to reduce serum levels of one or more inflammatory markers in the subject. 202. The method of embodiment 201, wherein the dose is effective to reduce serum levels of interleukin-6 ("IL-6"). 203. The method of embodiment 202, wherein the dose is effective to reduce serum levels of IL-6 by at least 20 pg / mL within 2 days of the first administration of the lipid-binding protein-based conjugate. 204. The method of embodiment 202, wherein the dose is effective to reduce serum levels of IL-6 by at least 30 pg / mL within 2 days of the first administration of the lipid-binding protein-based conjugate. 205. The method of any one of embodiments 202 to 204, wherein the dose is effective to reduce serum levels of IL-6 to 40 pg / mL within 2 days of the first administration of the lipid-binding protein-based complex. 206. The method of any one of embodiments 202 to 205, wherein the dose is effective to reduce serum levels of IL-6 by at least 20 pg / mL within 5 days of the first administration of the lipid-binding protein-based complex. 207. The method of any one of embodiments 202 to 205, wherein the dose is effective to reduce serum levels of IL-6 by at least 30 pg / mL within 5 days of the first administration of the lipid-binding protein-based complex. 208. The method of any one of embodiments 202 to 205, wherein the dose is effective to reduce serum levels of IL-6 by at least 40 pg / mL within 5 days of the first administration of the lipid-binding protein-based complex. 209. The method of any one of embodiments 202 to 208, wherein the dose is effective to reduce serum levels of IL-6 to 60 pg / mL within 5 days of the first administration of the lipid-binding protein-based complex. 210. The method of any one of embodiments 202 to 208, wherein the dose is effective to reduce serum levels of IL-6 to 50 pg / mL within 5 days of the first administration of the lipid-binding protein-based complex. 211. The method of any one of embodiments 201 to 210, wherein the dose is effective to reduce serum levels of C-reactive protein. 212. The method of any one of embodiments 201 to 211, wherein the dose is effective to reduce serum levels of D-dimer. 213. The method of any one of embodiments 201 to 212, wherein the dose is effective to reduce serum levels of ferritin. 214. The method of embodiment 213, wherein the dose is effective to reduce serum levels of ferritin by at least 200 ng / mL within 2 days of the first administration of the lipid-binding protein-based complex. 215. The method of embodiment 213, wherein the dose is effective to reduce serum levels of ferritin by at least 300 ng / mL within 2 days of the first administration of the lipid-binding protein-based complex. 216. The method of embodiment 213, wherein the dose is effective to reduce serum levels of ferritin by at least 400 ng / mL within 2 days of the first administration of the lipid-binding protein-based complex. 217. The method of any one of embodiments 213 to 216, wherein the dose is effective to reduce serum levels of ferritin to 700 ng / mL within 2 days of the first administration of the lipid binding protein-based complex. 218. The method of any one of embodiments 213 to 217, wherein the dose is effective to reduce serum levels of ferritin by at least 200 ng / mL within 5 days of the first administration of the lipid binding protein-based complex. 219. The method of any one of embodiments 213 to 217, wherein the dose is effective to reduce serum levels of ferritin by at least 300 ng / mL within 5 days of the first administration of the lipid binding protein-based complex. 220. The method of any one of embodiments 213 to 217, wherein the dose is effective to reduce serum levels of ferritin by at least 400 ng / mL within 5 days of the first administration of the lipid binding protein-based complex. 221. The method of any one of embodiments 213 to 220, wherein the dose is effective to reduce serum levels of ferritin to 700 ng / mL within 2 days of the first administration of the lipid binding protein-based complex. 222. The method of any one of embodiments 201 to 221, wherein the dose is effective to reduce serum levels of interleukin 8 (IL-8). 223. The method of embodiment 222, wherein the dose is effective to reduce serum levels of IL-8 by at least 100 pg / mL within 2 days of the first administration of the lipid-binding protein-based conjugate. 224. The method of embodiment 222, wherein the dose is effective to reduce serum levels of IL-8 by at least 150 pg / mL within 2 days of the first administration of the lipid-binding protein-based conjugate. 225. The method of any one of embodiments 222 to 224, wherein the dose is effective to reduce serum levels of IL-8 to 300 pg / mL within 2 days of the first administration of the lipid-binding protein-based complex. 226. The method of any one of embodiments 222 to 225, wherein the dose is effective to reduce serum levels of IL-8 by at least 100 pg / mL within 5 days of the first administration of the lipid-binding protein-based complex. 227. The method of any one of embodiments 222 to 225, wherein the dose is effective to reduce serum levels of IL-8 by at least 150 pg / mL within 5 days of the first administration of the lipid-binding protein-based complex. 228. The method of any one of embodiments 222 to 227, wherein the dose is effective to reduce serum levels of IL-8 to 300 pg / mL within 5 days of the first administration of the lipid-binding protein-based complex. 229. The method of any one of embodiments 201 to 228, wherein the dose is effective to reduce serum levels of granulocyte-macrophage colony-stimulating factor (GM-CSF). 230. The method of any one of embodiments 201 to 229, wherein the dose is effective to reduce serum levels of monocyte chemoattractant protein (MCP) 1. 231. The method of any one of embodiments 201 to 230, wherein the dose is effective to reduce serum levels of tumor necrosis factor alpha (TNF-α). 232. The method of any one of embodiments 201 to 231, wherein the dose is effective to reduce serum levels of KIM-1. 233. The method of any one of embodiments 201 to 232, wherein the dose is effective to reduce serum levels of one or more inflammatory markers from an elevated range to the normal range. 234. The method of any one of embodiments 201 to 233, wherein the dose is effective to reduce serum levels of one or more inflammatory markers by at least 20%, at least 40%, or at least 60%. 235. The method of any one of embodiments 1 to 234, wherein the subject has an endotoxin activity level of >0.6 prior to administration of the lipid-binding protein-based conjugate. 236. A method according to any one of embodiments 1 to 235, wherein the dose is effective to reduce endotoxin activity levels in the subject. 237. The method of any one of embodiments 1 to 236, wherein the dose is effective to transiently increase serum triglyceride levels. 238. The method of embodiment 237, wherein the transient increase is an increase for up to 9 days. 239. A method according to any one of embodiments 1 to 238, wherein the lipid-binding protein-based complex is a reconstituted HDL or HDL mimic. 240. A method according to any one of embodiments 1 to 238, wherein the lipid-binding protein-based complex is an Apomer or Cargomer. 241. A method according to any one of embodiments 1 to 240, wherein the lipid-binding protein-based complex comprises sphingomyelin. 242. A method according to any one of embodiments 1 to 241, wherein the lipid-binding protein-based complex comprises a negatively charged lipid. 243. The method of embodiment 242, wherein the negatively charged lipid is 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol) (DPPG) or a salt thereof. 244. The method of embodiment 239, wherein the lipid-binding protein-based complex is CER-001, CSL-111, CSL-112, CER-522, or ETC-216. 245. The method of embodiment 244, wherein the lipid-binding protein-based conjugate is CER-001. 246. A method according to any one of embodiments 1 to 245, in which the lipid-binding protein-based complex is administered systemically, optionally by infusion. 247. A method according to any one of embodiments 1 to 246, wherein the lipid-binding protein-based complex is administered until serum levels of one or more inflammatory markers are reduced. 248. The method of embodiment 247, wherein the lipid-binding protein-based complex is administered until serum levels of one or more inflammatory markers are reduced to the normal range. 249. The method of embodiment 247, wherein the lipid-binding protein-based complex is administered until the serum level of one or more inflammatory markers falls below the baseline level of one or more inflammatory markers measured prior to administration of the lipid-binding protein-based complex. 250. The method of any one of embodiments 1 to 249, wherein each individual dose of the lipid-binding protein-based conjugate administered is 4 to 40 mg / kg (protein weight basis). 251. The method of embodiment 250, wherein each individual dose of the lipid-binding protein-based conjugate is 4 to 30 mg / kg (protein weight basis). 252. The method of embodiment 250, wherein each individual dose of the lipid-binding protein-based conjugate is 15 to 25 mg / kg (protein weight basis). 253. The method of embodiment 250, wherein the individual dose of each of the lipid-binding protein-based conjugates is 10 to 30 mg / kg (protein weight basis). 254. The method of embodiment 250, wherein the individual dose of each of the lipid-binding protein-based conjugates is 10 to 20 mg / kg (protein weight basis). 255. The method of embodiment 250, wherein each individual dose of the lipid-binding protein-based conjugate is 5 mg / kg (protein weight basis). 256. The method of embodiment 250, wherein each individual dose of the lipid-binding protein-based conjugate is 10 mg / kg (protein weight basis). 257. The method of embodiment 250, wherein each individual dose of the lipid-binding protein-based conjugate is 15 mg / kg (protein weight basis). 258. The method of embodiment 250, wherein each individual dose of the lipid-binding protein-based conjugate is 20 mg / kg (protein weight basis). 259. The method of embodiment 250, wherein the individual dose of each of the lipid-binding protein-based conjugates is 5 to 15 mg / kg (protein weight basis). 260. The method of embodiment 250, wherein each individual dose of the lipid-binding protein-based conjugate is 10 to 20 mg / kg (protein weight basis). 261. The method of embodiment 250, wherein each individual dose of the lipid-binding protein-based conjugate is 15 to 25 mg / kg (protein weight basis). 262. The method of any one of embodiments 1 to 261, wherein the dose is administered according to an induction regimen, optionally followed by a consolidation therapy regimen. 263. The method of embodiment 262, wherein the induction regimen comprises administering the lipid-binding protein-based conjugate once a day or twice a day. 264. The method of embodiment 262 or embodiment 263, wherein the consolidation therapy regimen comprises administering a lipid-binding protein-based conjugate once a day or once every two days. 265. The method of any one of embodiments 1 to 264, wherein the subject is not treated with a maintenance regimen. 266. The method of any one of embodiments 262 to 265, wherein the consolidation therapy regimen comprises administering one or more doses of a lipid-binding protein-based complex to the subject one or more days after administration of the final dose of the induction regimen. 267. The method of embodiment 266, wherein the first dose of the lipid-binding protein-based conjugate administered during the consolidation therapy regimen is administered more than two days after administration of the final dose of the induction regimen. 268. The method of embodiment 266, wherein the first dose of the lipid-binding protein-based conjugate administered during the consolidation therapy regimen is administered 3 days or more after administration of the final dose of the induction regimen. 269. The method of embodiment 268, wherein the first dose of the lipid-binding protein-based conjugate administered during the consolidation therapy regimen is administered 3 days after administration of the final dose of the induction regimen. 270. The method of any one of embodiments 262 to 269, comprising an induction regimen comprising administration of a lipid-binding protein-based conjugate twice daily on days 1, 2, and 3, and a consolidation therapy regimen comprising two doses of a lipid-binding protein-based conjugate on day 6. 271. The method of any one of embodiments 262 to 270, wherein each individual dose of lipid-binding protein-based conjugate administered in the induction regimen is 4 to 40 mg / kg (protein weight basis). 272. The method of any one of embodiments 262 to 271, wherein each individual dose of lipid-binding protein-based conjugate administered in the induction regimen is 4 to 30 mg / kg (protein weight basis). 273. The method of any one of embodiments 262 to 271, wherein each individual dose of lipid-binding protein-based conjugate administered in the induction regimen is 15 to 25 mg / kg (protein weight basis). 274. The method of any one of embodiments 262 to 271, wherein each individual dose of lipid-binding protein-based conjugate administered in the induction regimen is 10 to 30 mg / kg (protein weight basis). 275. The method of any one of embodiments 262 to 271, wherein each individual dose of lipid-binding protein-based conjugate administered in the induction regimen is 10 to 20 mg / kg (protein weight basis). 276. The method of any one of embodiments 262 to 271, wherein each individual dose of the lipid-binding protein-based conjugate administered in the induction regimen is 5 mg / kg (protein weight basis). 277. The method of any one of embodiments 262 to 271, wherein each individual dose of the lipid-binding protein-based conjugate administered in the induction regimen is 10 mg / kg (protein weight basis). 278. The method of any one of embodiments 262 to 271, wherein each individual dose of the lipid-binding protein-based conjugate administered in the induction regimen is 15 mg / kg (protein weight basis). 279. The method of any one of embodiments 262 to 271, wherein each individual dose of the lipid-binding protein-based conjugate administered in the induction regimen is 20 mg / kg (protein weight basis). 280. The method of any one of embodiments 262 to 279, wherein the dose of the lipid-binding protein-based conjugate administered in the consolidation therapy regimen is 5 to 15 mg / kg (protein weight basis). 281. The method of any one of embodiments 262 to 279, wherein the dose of the lipid-binding protein-based conjugate administered in the consolidation therapy regimen is 10 to 20 mg / kg (protein weight basis). 282. The method of any one of embodiments 262 to 279, wherein the dose of the lipid-binding protein-based conjugate administered in the consolidation therapy regimen is 15 to 25 mg / kg (protein weight basis). 283. The method of any one of embodiments 262 to 279, wherein the dose of the lipid-binding protein-based conjugate administered in the consolidation therapy regimen is 5 mg / kg (protein weight basis). 284. The method of any one of embodiments 262 to 279, wherein the dose of the lipid-binding protein-based conjugate administered in the consolidation therapy regimen is 10 mg / kg (protein weight basis). 285. The method of any one of embodiments 262 to 279, wherein the dose of the lipid-binding protein-based conjugate administered in the consolidation therapy regimen is 15 mg / kg (protein weight basis). 286. The method of any one of embodiments 1 to 285, wherein each individual dose of the lipid-binding protein-based conjugate administered is between 300 mg and 4000 mg (protein weight basis). 287. The method of embodiment 286, wherein each individual dose of the lipid-binding protein-based conjugate administered is between 300 mg and 3000 mg (protein weight basis). 288. The method of embodiment 286, wherein each individual dose of the lipid-binding protein-based conjugate administered is between 300 mg and 1500 mg (protein weight basis). 289. The method of embodiment 286, wherein each individual dose of the lipid-binding protein-based conjugate administered is between 400 mg and 4000 mg (protein weight basis). 290. The method of embodiment 286, wherein each individual dose of the lipid-binding protein-based conjugate administered is between 400 mg and 1500 mg (protein weight basis). 291. The method of embodiment 286, wherein each individual dose of the lipid-binding protein-based conjugate administered is 500 mg to 1200 mg (protein weight basis). 292. The method of embodiment 286, wherein each individual dose of the lipid-binding protein-based conjugate administered is 500 mg to 1000 mg (protein weight basis). 293. The method of embodiment 286, wherein each individual dose of the lipid-binding protein-based conjugate administered is between 600 mg and 3000 mg (protein weight basis). 294. The method of embodiment 286, wherein each individual dose of the lipid-binding protein-based conjugate administered is between 800 mg and 3000 mg (protein weight basis). 295. The method of embodiment 286, wherein each individual dose of the lipid-binding protein-based conjugate administered is between 1000 mg and 2400 mg (protein weight basis). 296. The method of embodiment 286, wherein each individual dose of the lipid-binding protein-based conjugate administered is 1000 mg to 2000 mg (protein weight basis). 297. The method of any one of embodiments 1 to 296, wherein the high dose of the lipid-binding protein-based complex is 600 mg to 40 g (protein weight basis). 298. The method of any one of embodiments 1 to 296, wherein the high dose of the lipid-binding protein-based complex is 3 g to 35 g (protein weight basis). 299. The method of any one of embodiments 1 to 296, wherein the high dose of the lipid-binding protein-based complex is 5 g to 30 g (protein weight basis). 300. A method according to any one of embodiments 1 to 299, wherein the lipid-binding protein-based complex is administered by infusion. 301. The method of embodiment 300, in which each individual dose is administered over a period of 1 to 24 hours. 302. The method of embodiment 301, in which each individual dose is administered over the course of a 24-hour period. 303. The method of any one of embodiments 1 to 302, further comprising administering to the subject an antihistamine prior to each individual dose. 304. The method of embodiment 303, wherein the antihistamine comprises dexchlorphenylamine or hydroxyzine. 305. The method of any one of embodiments 1 to 304, wherein the subject is undergoing or has undergone one or more additional therapies, and / or further comprising administering one or more additional therapies to the subject. 306. The method of embodiment 305, wherein the one or more additional treatments include a standard treatment. 307. The method of embodiment 305 or embodiment 306, wherein the subject has an infection and the one or more additional treatments comprises antibiotic treatment. 308. The method of embodiment 305 or embodiment 306, wherein the one or more additional therapies include hemodynamic support. 309. The method of any one of embodiments 305 to 308, wherein the one or more additional therapies comprise one or more anti-IL-6 agents. 310. The method of embodiment 309, wherein the one or more anti-IL-6 agents comprise tocilizumab, siltuximab, olokizumab, ersilimomab, BMS-945429, sirukumab, revilimab, CPSI-2364, or a combination thereof. 311. The method of embodiment 310, wherein the one or more anti-IL-6 agents comprises tocilizumab. 312. The method of any one of embodiments 305 to 311, wherein the one or more additional treatments comprise one or more corticosteroids. 313. The method of embodiment 312, wherein the one or more corticosteroids comprise methylprednisolone, dexamethasone, or a combination thereof. 314. The method of any one of embodiments 305 to 313, wherein the subject has or has had a COVID-19 infection and the one or more additional treatments include antibodies from recovered COVID-19 patients. 315. The method of any one of embodiments 305 to 314, wherein the subject has or has had a COVID-19 infection and the one or more additional treatments comprises an antibody against the spike protein of COVID-19. 316. The method of any one of embodiments 305 to 315, wherein the subject has or has had a COVID-19 infection and the one or more additional treatments include one or more antiviral agents. 317. The method of embodiment 316, wherein the one or more antiviral agents comprises lopinavir. 318. The method of embodiment 316 or embodiment 317, wherein the one or more antiviral agents comprises remdesivir. 319. The method of any one of embodiments 316 to 318, wherein the one or more antiviral agents comprises danoprevir. 320. The method of any one of embodiments 316 to 319, wherein the one or more antiviral agents comprises galidesivir. 321. The method of any one of embodiments 316 to 320, wherein the one or more antiviral agents comprises darunavir. 322. The method of any one of embodiments 316 to 321, wherein the one or more antiviral agents comprises ritonavir. 323. The method of any one of embodiments 305 to 322, wherein the subject has or has had a COVID-19 infection and the one or more additional treatments comprises chloroquine or hydroxychloroquine. 324. The method of any one of embodiments 305 to 323, wherein the subject has or has had a COVID-19 infection and the one or more additional treatments comprises azithromycin. 325. The method of any one of embodiments 305 to 324, wherein the subject has or has had a COVID-19 infection and the one or more additional treatments comprises interferon. 326. The method of embodiment 325, wherein the interferon is interferon alpha. 327. The method of embodiment 325, wherein the interferon is interferon beta. 328. The method of any one of embodiments 325 to 327, wherein the interferon is pegylated. 329. The method of any one of embodiments 1 to 328, wherein the lipid-binding protein-based conjugate is CER-001. 330. The method of embodiment 329, wherein CER-001 is a lipoprotein complex comprising ApoA-I and phospholipids in a ratio of 1:2.7+ / -20% ApoA-I weight:total phospholipid weight and phospholipids sphingomyelin and DPPG in a weight:weight ratio of 97:3+ / -20% sphingomyelin:DPPG. 331. The method of embodiment 329, wherein CER-001 is a lipoprotein complex comprising ApoA-I and phospholipids in a ratio of 1:2.7+ / -10% ApoA-I weight:total phospholipid weight and phospholipids sphingomyelin and DPPG in a weight:weight ratio of 97:3+ / -10% sphingomyelin:DPPG. 332. The method of embodiment 329, wherein CER-001 is a lipoprotein complex comprising ApoA-I and phospholipids in a ratio of ApoA-I weight:total phospholipid weight of 1:2.7 and the phospholipids sphingomyelin and DPPG in a weight:weight ratio of sphingomyelin:DPPG of 97:3. 333. The method of any one of embodiments 330 to 332, wherein ApoA-I has the amino acid sequence of amino acids 25 to 267 of SEQ ID NO:2. 334. The method of any one of embodiments 330 to 333, wherein ApoA-I is recombinantly expressed. 335. The method of any one of embodiments 330 to 334, wherein CER-001 comprises natural sphingomyelin. 336. The method of embodiment 335, wherein the natural sphingomyelin is chicken egg sphingomyelin. 337. The method of any one of embodiments 330 to 334, wherein CER-001 comprises synthetic sphingomyelin. 338. The method of embodiment 337, wherein the synthetic sphingomyelin is palmitoyl sphingomyelin. 339. The method of any one of embodiments 329 to 338, wherein CER-001 is administered in the form of a formulation in which CER-001 is at least 95% homogeneous. 340. The method of embodiment 339, wherein CER-001 is administered in the form of a formulation in which CER-001 is at least 97% homogeneous. 341. The method of embodiment 339, wherein CER-001 is administered in the form of a formulation in which CER-001 is at least 98% homogeneous. 342. The method of embodiment 339, wherein CER-001 is administered in the form of a formulation in which CER-001 is at least 99% homogeneous.

[0345] Various aspects of the disclosure are described in the embodiments set forth in the numbered paragraphs of Group 2 below.

[0346] Group 2: 1. A method of treating a subject having one or more symptoms associated with leukocytosis, comprising administering to the subject a dose of an apolipoprotein AI ("ApoA-I") formulation comprising ApoA-I and one or more lipids, wherein the ApoA-I and the lipids are in the form of a lipoprotein complex that is effective for lowering the subject's white blood cell count and / or ameliorating one or more of the one or more symptoms associated with leukocytosis. 2. A method for treating a subject having endothelial dysfunction, comprising administering to the subject an ApoA-I formulation comprising ApoA-I and one or more lipids, wherein the ApoA-I and lipids are in the form of a lipoprotein complex. 3. A method for treating a subject having or at risk of developing carditis, comprising administering to the subject an ApoA-I formulation comprising ApoA-I and one or more lipids, wherein the ApoA-I and lipids are in the form of a lipoprotein complex. 4. A method of treating a subject having or at risk of developing leukocytosis, comprising administering to the subject a dose of an ApoA-I formulation comprising ApoA-I and one or more lipids, wherein the ApoA-I and the lipids are in the form of a lipoprotein complex that is effective to lower the subject's white blood cell count. 5. A method of treating a subject experiencing or who has experienced an acute coronary syndrome or stroke, comprising administering to the subject an ApoA-I formulation comprising ApoA-I and one or more lipids, wherein the ApoA-I and lipids are in the form of a lipoprotein complex. 6. The method of any one of embodiments 1 to 5, wherein ApoA-I has the amino acid sequence of amino acids 25 to 267 of SEQ ID NO:2. 7. The method of any one of embodiments 1 to 6, wherein ApoA-I is recombinantly expressed. 8. The method of any one of embodiments 1 to 7, wherein the one or more lipids comprise neutral lipids. 9. The method of embodiment 8, wherein the neutral lipid comprises sphingomyelin. 10. The method of embodiment 9, wherein the neutral lipids consist of sphingomyelin. 11. The method of embodiment 9 or embodiment 10, wherein the sphingomyelin comprises native sphingomyelin. 12. The method of embodiment 11, wherein the natural sphingomyelin is chicken egg sphingomyelin. 13. The method of embodiment 9 or embodiment 10, wherein the sphingomyelin comprises synthetic sphingomyelin. 14. The method of embodiment 13, wherein the synthetic sphingomyelin is palmitoyl sphingomyelin. 15. The method of any one of embodiments 1 to 10, wherein the one or more lipids further comprise a negatively charged lipid. 16. The method of embodiment 15, wherein the negatively charged lipid comprises 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] ("DPPG") or a salt thereof. 17. The method of embodiment 16, wherein the negatively charged lipid consists of DPPG or a salt thereof. 18. The method of any one of embodiments 11 to 17, wherein the molar ratio of negatively charged lipid components to neutral lipids to ApoA-I in the formulation is 2-6:90-120:1. 19. The method of any one of embodiments 8 to 18, wherein the lipids consist of 95-99% by weight of neutral phospholipids and 1-5% by weight of negatively charged phospholipids. 20. The method of embodiment 19, wherein the lipids consist of 96-98% by weight of neutral phospholipids and 2-4% by weight of negatively charged phospholipids. 21. The method of embodiment 20, wherein the lipids consist of 97% by weight neutral phospholipids and 3% by weight negatively charged phospholipids. 22. A method according to any one of embodiments 1 to 21, having a ratio of ApoA-I to lipid ranging from 1:2 to 1:3 by weight. 23. The method of embodiment 22, having an ApoA-I to lipid ratio of about 1:2.7 by weight. 24. The method of any one of embodiments 1 to 23, wherein the lipoprotein complex is at least 95% homogeneous, as reflected by a single peak in gel permeation chromatography. 25. An apolipoprotein AI ("ApoA-I") formulation comprising ApoA-I and one or more lipids for use in treating a subject having one or more conditions associated with leukocytosis, wherein the ApoA-I and lipid are in the form of a lipoprotein complex. 26. An apolipoprotein AI ("ApoA-I") formulation comprising ApoA-I and one or more lipids for use in treating a subject with endothelial dysfunction, wherein the ApoA-I and lipids are in the form of a lipoprotein complex. 27. An apolipoprotein AI ("ApoA-I") formulation comprising ApoA-I and one or more lipids for use in treating a subject having or at risk of developing carditis, wherein the ApoA-I and lipid are in the form of a lipoprotein complex. 28. An apolipoprotein AI ("ApoA-I") formulation comprising ApoA-I and one or more lipids for use in treating a subject having or at risk of developing leukocytosis, wherein the ApoA-I and lipid are in the form of a lipoprotein complex. 29. An apolipoprotein AI ("ApoA-I") formulation comprising ApoA-I and one or more lipids, for use in treating a subject experiencing or who has experienced an acute coronary syndrome or stroke, wherein the ApoA-I and lipids are in the form of a lipoprotein complex. 30. A formulation for use according to any one of embodiments 25 to 29, wherein ApoA-I has the amino acid sequence of amino acids 25 to 267 of SEQ ID NO:2. 31. A formulation for use according to any one of embodiments 25 to 30, wherein ApoA-I is recombinantly expressed. 32. A formulation for use according to any one of embodiments 25 to 31, wherein the one or more lipids comprises a neutral lipid. 33. The formulation for use according to embodiment 32, wherein the neutral lipid comprises sphingomyelin. 34. The formulation for use according to embodiment 33, wherein the neutral lipid comprises sphingomyelin. 35. The formulation for use according to embodiment 33 or embodiment 34, wherein the sphingomyelin comprises natural sphingomyelin. 36. The formulation for use according to embodiment 35, wherein the natural sphingomyelin is chicken egg sphingomyelin. 37. The formulation for use according to embodiment 33 or embodiment 34, wherein the sphingomyelin comprises synthetic sphingomyelin. 38. The formulation for use according to embodiment 37, wherein the synthetic sphingomyelin is palmitoyl sphingomyelin. 39. A formulation for use according to any one of embodiments 25 to 34, wherein the one or more lipids further comprise a negatively charged lipid. 40. The formulation for use according to embodiment 39, wherein the negatively charged lipid comprises 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] ("DPPG") or a salt thereof. 41. The formulation for use according to embodiment 40, wherein the negatively charged lipid consists of DPPG or a salt thereof. 42. A formulation for use according to any one of embodiments 35 to 41, wherein the molar ratio of negatively charged lipid components to neutral lipids to ApoA-I in the formulation is 2-6:90-120:1. 43. A formulation for use according to any one of embodiments 32 to 42, wherein the lipids consist of 95 to 99% by weight of neutral phospholipids and 1 to 5% by weight of negatively charged phospholipids. 44. The method of embodiment 43, wherein the lipids consist of 96-98% by weight of neutral phospholipids and 2-4% by weight of negatively charged phospholipids. 45. A formulation for use according to embodiment 44, wherein the lipids consist of 97% by weight of neutral phospholipids and 3% by weight of negatively charged phospholipids. 46. ​​A formulation for use according to any one of embodiments 25 to 45, having a ratio of ApoA-I to lipid in the range of 1:2 to 1:3 by weight. 47. A formulation for use according to embodiment 46, having an ApoA-I to lipid ratio of about 1:2.7 by weight. 48. A formulation for use according to any one of embodiments 25 to 47, wherein the lipoprotein complex is at least 95% homogeneous, as reflected by a single peak in gel permeation chromatography. 49. An apolipoprotein AI ("ApoA-I") formulation comprising ApoA-I and one or more lipids, for use in the manufacture of a medicament for one or more conditions associated with leukocytosis, wherein the ApoA-I and the lipid are in the form of a lipoprotein complex. 50. An apolipoprotein AI ("ApoA-I") formulation comprising ApoA-I and one or more lipids, for use in the manufacture of a medicament for endothelial dysfunction, wherein the ApoA-I and the lipid are in the form of a lipoprotein complex. 51. An apolipoprotein AI ("ApoA-I") formulation comprising ApoA-I and one or more lipids, for use in the manufacture of a medicament for carditis or the risk of developing carditis, wherein the ApoA-I and the lipid are in the form of a lipoprotein complex. 52. An apolipoprotein AI ("ApoA-I") formulation comprising ApoA-I and one or more lipids, for use in the manufacture of a medicament for leukocytosis or the risk of developing leukocytosis, wherein the ApoA-I and the lipid are in the form of a lipoprotein complex. 53. An apolipoprotein AI ("ApoA-I") formulation comprising ApoA-I and one or more lipids, for use in the manufacture of a medicament for acute coronary syndrome or stroke, wherein the ApoA-I and lipid are in the form of a lipoprotein complex. 54. A formulation for use according to any one of embodiments 49 to 53, wherein ApoA-I has the amino acid sequence of amino acids 25 to 267 of SEQ ID NO:2. 55. A formulation for use according to any one of embodiments 49 to 54, wherein ApoA-I is recombinantly expressed. 56. A formulation for use according to any one of embodiments 49 to 55, wherein the one or more lipids comprise neutral lipids. 57. A formulation for use according to embodiment 56, wherein the neutral lipid comprises sphingomyelin. 58. A formulation for use according to embodiment 57, wherein the neutral lipid comprises sphingomyelin. 59. A formulation for use according to embodiment 57 or embodiment 58, wherein the sphingomyelin comprises natural sphingomyelin. 60. A formulation for use according to embodiment 59, wherein the natural sphingomyelin is chicken egg sphingomyelin. 61. A formulation for use according to embodiment 57 or embodiment 58, wherein the sphingomyelin comprises synthetic sphingomyelin. 62. A formulation for use according to embodiment 61, wherein the synthetic sphingomyelin is palmitoyl sphingomyelin. 63. A formulation for use according to any one of embodiments 49 to 58, wherein the one or more lipids further comprise a negatively charged lipid. 64. The formulation for use according to embodiment 63, wherein the negatively charged lipid comprises 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] ("DPPG") or a salt thereof. 65. The formulation for use according to embodiment 64, wherein the negatively charged lipid consists of DPPG or a salt thereof. 66. A formulation for use according to any one of embodiments 59 to 65, wherein the molar ratio of negatively charged lipid components to neutral lipids to ApoA-I in the formulation is 2-6:90-120:1. 67. A formulation for use according to any one of embodiments 56 to 66, wherein the lipids consist of 95 to 99% by weight of neutral phospholipids and 1 to 5% by weight of negatively charged phospholipids. 68. The method of embodiment 67, wherein the lipids consist of 96-98% by weight of neutral phospholipids and 2-4% by weight of negatively charged phospholipids. 69. A formulation for use according to embodiment 68, wherein the lipids consist of 97% by weight of neutral phospholipids and 3% by weight of negatively charged phospholipids. 70. A formulation for use according to any one of embodiments 49 to 69, having a ratio of ApoA-I to lipid in the range of 1:2 to 1:3 by weight. 71. A formulation for use according to embodiment 70, having an ApoA-I to lipid ratio of about 1:2.7 by weight. 72. A formulation for use according to any one of embodiments 49 to 71, wherein the lipoprotein complex is at least 95% homogeneous, as reflected by a single peak in gel permeation chromatography. 73. A method or formulation for use according to any one of embodiments 1 to 72, wherein the dose is administered twice a day. 74. A method or formulation for use according to any one of embodiments 1 to 73, wherein the dose is administered for 5 days. 75. The method or formulation for use according to any one of embodiments 1 to 74, wherein the dose is effective to reduce the subject's white blood cell count by at least 2000 WBC / microliter within 5 days of the first administration of the lipid binding protein-based conjugate. 76. The method or formulation for use of any one of embodiments 1 to 75, wherein the dose is effective to reduce the subject's white blood cell count by at least 3000 WBC / microliter within 5 days of the first administration of the lipid binding protein-based conjugate.

Claims

1. A lipid-binding protein-based complex used in a method for treating a subject having one or more symptoms associated with leukocytosis, comprising administering to the subject a dose of the lipid-binding protein-based complex that is effective in reducing the subject's white blood cell count and / or alleviating one or more of the one or more symptoms associated with leukocytosis.

2. The complex according to claim 1, wherein one or more of the symptoms include fever, bleeding or bruising, sweating, pain or stinging in the legs, arms, or abdomen, one or more visual problems, blurred thinking, loss of appetite, and / or difficulty breathing (e.g., shortness of breath, subnormal blood oxygen levels).

3. The complex according to claim 1 or 2, wherein the subject has or is at risk of developing leukocytosis caused by inflammation, infection, leukocyte disorder, physical stress, emotional stress, drugs, or allergic reactions.

4. The aforementioned subject is, (a) Having an infectious disease, wherein the infectious disease is a viral infection, bacterial infection, fungal infection, or parasitic infection, and the viral infection may be a coronavirus infection, and the coronavirus infection may be COVID-19. (b) Do you have diabetes? (c) Having a leukocyte disorder, (d) Having leukocytosis, (e) The complex according to claim 1, having a white blood cell count of more than 11 × 10⁹ cells / L.

5. A lipid-binding protein-based complex used in a method for treating a subject having or at risk of developing carditis, comprising administering a predetermined dose of the lipid-binding protein-based complex to the subject, wherein the carditis may be myocarditis and / or pericarditis.

6. The composite according to claim 5, wherein the subject has an infectious disease, for example, a viral infection or a bacterial infection, and the viral infection may be a coronavirus infection, and the coronavirus infection may be COVID-19.

7. The composite according to claim 5 or 6, wherein the subject has a risk of cardiac inflammation.

8. The complex according to claim 7, wherein the dose of the complex based on the lipid-binding protein is effective in preventing or reducing the severity of the carditis.

9. A lipid-binding protein-based complex used in a method for treating a subject having or at risk of developing leukocytosis, comprising administering to the subject a dose of the lipid-binding protein-based complex that is effective in reducing the subject's white blood cell count.

10. The complex according to claim 1, wherein the lipid-binding protein-based complex is CER-001.

11. an apolipoprotein A-I ("ApoA-I") formulation comprising ApoA-I and one or more lipids, used in a method for treating subjects having one or more symptoms associated with leukocytosis.

12. an apolipoprotein A-I ("ApoA-I") formulation comprising ApoA-I and one or more lipids, used in a method for treating subjects having or at risk of developing carditis, wherein the ApoA-I and the lipids are in the form of lipoprotein complexes.

13. an apolipoprotein A-I ("ApoA-I") formulation comprising ApoA-I and one or more lipids, used in a method for treating subjects who have or are at risk of developing leukocytosis.

14. The formulation according to claim 11, wherein ApoA-I has the amino acid sequence of amino acids 25 to 267 of SEQ ID NO:

2.

15. The formulation according to claim 11, wherein ApoA-I is expressed by recombinant DNA.

16. The formulation according to claim 11, wherein one or more of the lipids include a triglyceride.

17. The formulation according to claim 16, wherein the aforementioned triglycerides contain sphingomyelin.

18. The formulation according to claim 11, wherein the synthetic sphingomyelin is palmitoyl sphingomyelin.

19. The formulation according to claim 19, wherein the one or more lipids further comprise loaded electrolipids.

20. The formulation according to claim 19, wherein the loaded electrolipid comprises 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] ("DPPG") or a salt thereof.

21. The formulation according to claim 16, wherein the lipids consist of 95 to 99% by weight of neutral phospholipids and 1 to 5% by weight of loaded electrophospholipids.

22. The formulation according to claim 21, wherein the lipids consist of 96 to 98% by weight of neutral phospholipids and 2 to 4% by weight of loaded electrophospholipids.

23. The formulation according to claim 22, wherein the lipids consist of 97% by weight of neutral phospholipids and 3% by weight of loaded electrophospholipids.

24. The formulation according to claim 11, having an ApoA-I to lipid ratio in the range of 1:2 to 1:3 by weight.

25. The formulation according to claim 24, having an ApoA-I to lipid ratio of approximately 1:2.7 by weight.

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