Method for treating a hyperinflammatory state using a complex based on a lipid-binding protein

Abstract

A method for treating hyperinflammatory conditions such as hemophagocytic lymphohistiocytosis (HLH), dengue hemorrhagic fever, and dengue shock syndrome using a complex based on a lipid-binding protein.

Description

Technical Field

[0001] 1. Cross - Reference to Related Applications This application claims the benefit of priority of U.S. Application No. 63 / 351,129, filed on June 10, 2022, the content of which is incorporated herein by reference in its entirety.

[0002] 2. Sequence Listing This application includes a sequence listing submitted electronically in XML format, which is incorporated herein by reference in its entirety. The XML sequence listing created on June 1, 2023 is named CRN - 050WO_SL.xml and its size is 3,275 bytes.

Background Art

[0003] 3. Background Art In some studies, it has been reported that ApoA - I and HDL are not as abundant in COVID - 19 patients, especially in the most severe forms, and that HDL derived from COVID - 19 patients is not as protective in endothelial cells exposed to inflammatory triggers and does not protect endothelial cells from apoptosis. A decrease in serum levels of ApoA - I can increase both the risk of developing COVID - 19 and the risk of COVID - 19 becoming a severe form. Such a decrease in ApoA - I or HDL is a common finding in cytokine storms and is also observed in virus - induced and familial hemophagocytic lymphohistiocytosis (HLH), as well as dengue shock syndrome.

[0004] Current treatment methods for such highly inflammatory conditions are often insufficient or not optimal. Therefore, new treatment methods for highly inflammatory conditions, such as virus - induced highly inflammatory conditions, are needed.

Summary of the Invention

[0005] 4. Summary The present disclosure provides methods for treating a subject having or at risk of having an inflammatory condition, such as hemophagocytic lymphohistiocytosis (HLH), dengue hemorrhagic fever, and dengue shock syndrome. In some embodiments, the subject has hyperinflammation characterized by severe inflammation with a cytokine storm.

[0006] In some embodiments, the subject is treated with a high-dose lipid-bound protein-based complex. The high dose is typically higher than the dose that would be used to treat chronic conditions such as familial hypercholesterolemia. The high dose is typically administered over a relatively short period of time, such as over a period of 1 day to 2 weeks, and typically includes multiple administrations of the lipid-bound protein-based complex, such as 2 to 10 individual doses. The individual doses may be separated by less than 1 day (e.g., twice daily administration) or by more than 1 day (e.g., once daily administration).

[0007] In some embodiments of the methods of the present disclosure, the lipid-bound protein-based complex comprises sphingomyelin and / or a charged lipid, such as CER-001. CER-001 is a charged lipoprotein complex that comprises recombinant human ApoA-I, sphingomyelin (SM), and 1,2-dihexadecanoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (dipalmitoyl phosphatidyl-glycerol, DPPG). This mimics native nascent discoidal pre-beta HDL, the form that HDL particles take before acquiring cholesterol. Without being bound by theory, CER-001 treatment is thought to reduce serum levels of inflammatory cytokines, such as IL-6, and thereby provide a clinical benefit to a subject having an inflammatory condition described herein, such as a subject having or at risk of having a virus-induced hyperinflammatory condition.

[0008] In one aspect, the present disclosure provides a method of treating a subject having or at risk of having HLH, the method comprising administering to the subject a lipid-binding protein-based complex (e.g., CER-001).

[0009] In one aspect, the present disclosure provides a method of treating a subject having or at risk of having familial HLH, the method comprising administering to the subject a lipid-binding protein-based complex (e.g., CER-001).

[0010] In one aspect, the present disclosure provides a method of treating a subject having or at risk of having HLH subsequent to a malignant disease (e.g., acute leukemia or lymphoma) or a non-malignant disease (e.g., an autoimmune disease or an infectious disease), the method comprising administering to the subject a lipid-binding protein-based complex (e.g., CER-001).

[0011] In one aspect, the present disclosure provides a method of treating a subject having or at risk of having virus-induced HLH, the method comprising administering to the subject a lipid-binding protein-based complex (e.g., CER-001).

[0012] In another aspect, the present disclosure provides a method of treating a subject having dengue infection, the method comprising administering to the subject a lipid-binding protein-based complex (e.g., CER-001).

[0013] In another aspect, the present disclosure provides a method of treating a subject having or at risk of having dengue hemorrhagic fever, the method comprising administering to the subject a lipid-binding protein-based complex (e.g., CER-001).

[0014] In another aspect, the present disclosure provides a method of treating a subject having or at risk of having dengue shock syndrome, the method comprising administering to the subject a lipid-binding protein-based complex (e.g., CER-001).

[0015] In another aspect, the present disclosure provides a method of treating a subject having a herpes simplex infection, the method comprising administering to the subject a lipid-bound protein-based complex (e.g., CER-001).

[0016] In some aspects, the present disclosure provides a dosing regimen for a lipid-bound protein-based treatment (e.g., CER-001 treatment) performed on a subject as described herein.

[0017] The dosing regimens of the present disclosure generally require multiple administrations of CER-001 to the subject (e.g., administered daily or twice daily). CER-001 treatment can be continued for a pre-determined period, e.g., within one week (e.g., 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, or 7 days), or for a period longer than one week (e.g., 2 weeks). Alternatively, administration of CER-001 to the subject can be continued until one or more symptoms of a condition (e.g., acute inflammation or cytokine release syndrome (CRS)) are reduced, or until the serum levels of one or more inflammatory markers are decreased, e.g., to normal levels, or decreased compared to baseline measurements obtained prior to initiating CER-001 treatment. In the case of a subject having an infectious disease (e.g., a viral infection), the treatment can, in some embodiments, be continued until the subject recovers from the infectious disease.

[0018] The dosing regimens of the present disclosure may involve administering a lipid-bound protein-based complex (e.g., CER-001) to the subject according to an initial "induction" regimen, optionally followed by administering a lipid-bound protein-based complex to the subject according to a "consolidation therapy" regimen.

[0019] The induction regimen typically includes administering multiple doses of a lipid-bound protein-based complex (e.g., CER-001) to the subject, e.g., 6 doses over 3 days.

[0020] The consolidation therapy regimen typically involves administering one or more doses of a lipid-bound protein-based complex (e.g., CER-001) to a subject, once or multiple times, after the final dose of the induction regimen, for example, 1 day or multiple 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 include administration of a lipid-bound protein-based complex (e.g., CER-001) to a subject according to the induction regimen on days 1, 2, and 3, and administration of a lipid-bound protein-based complex to the subject according to the consolidation therapy regimen on day 6. In some embodiments, the consolidation therapy regimen includes two doses of a lipid-bound protein-based complex.

[0021] In certain embodiments, the present disclosure provides the following dosing regimen: - On days 1, 2, and 3, two doses per day (induction regimen), optionally followed by - After day 4, two subsequent doses (consolidation therapy regimen) for treating a subject having or at risk of having HLH (e.g., virus-induced HLH, familial HLH, or HLH subsequent to acute leukemia or lymphoma), having dengue infection, having or at risk of having dengue hemorrhagic fever, having or at risk of having dengue shock syndrome, or having herpes simplex infection, with a lipid-bound protein-based complex (e.g., CER-001). In some embodiments, the regimen is - On days 1, 2, and 3, two doses per day (induction regimen), followed by - On day 6, two doses (consolidation therapy regimen) and includes.

[0022] In certain aspects, the lipid-bound protein-based complex (e.g., CER-001) is administered in combination with standard therapy for the subject's disease or condition.

[0023] In certain embodiments, an antihistamine (e.g., dexchlorpheniramine, hydroxyzine, diphenhydramine, cetirizine, fexofenadine, or loratadine) can be administered prior to administration of a complex based on a lipid-binding protein (e.g., CER-001). The antihistamine can reduce the likelihood of an allergic reaction. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] 5. BRIEF DESCRIPTION OF THE DRAWINGS

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Mode for Carrying Out the Invention

[0025] 6. Detailed Description The present disclosure provides a method for treating a subject having or at risk of having an inflammatory condition, such as hemophagocytic lymphohistiocytosis (HLH), dengue hemorrhagic fever, and dengue shock syndrome, using a lipid-binding protein-based complex.

[0026] In some embodiments, the method includes administering a high dose of a lipid-binding protein-based complex.

[0027] In one aspect, the present disclosure provides a method for treating a subject having or at risk of having HLH, the method comprising administering to the subject a lipid-binding protein-based complex (e.g., CER-001).

[0028] In one aspect, the present disclosure provides a method for treating a subject having or at risk of having familial HLH, the method comprising administering to the subject a lipid-binding protein-based complex (e.g., CER-001).

[0029] In one aspect, the present disclosure provides a method for treating a subject having or at risk of having HLH subsequent to a malignant disease (e.g., acute leukemia or lymphoma), or a non-malignant disease (e.g., an autoimmune disease or an infectious disease), the method comprising administering to the subject a lipid-binding protein-based complex (e.g., CER-001).

[0030] In one aspect, the present disclosure provides a method of treating a subject having or at risk of having virus-induced HLH, the method comprising administering to the subject a lipid-binding protein-based complex (e.g., CER-001).

[0031] In another aspect, the present disclosure provides a method of treating a subject having dengue infection (e.g., a subject having dengue fever, dengue hemorrhagic fever, or dengue shock syndrome), the method comprising administering to the subject a lipid-binding protein-based complex (e.g., CER-001).

[0032] In another aspect, the present disclosure provides a method of treating a subject having or at risk of having dengue hemorrhagic fever, the method comprising administering to the subject a lipid-binding protein-based complex (e.g., CER-001).

[0033] In another aspect, the present disclosure provides a method of treating a subject having or at risk of having dengue shock syndrome, the method comprising administering to the subject a lipid-binding protein-based complex (e.g., CER-001).

[0034] In another aspect, the present disclosure provides a method of treating a subject having herpes simplex infection, the method comprising administering to the subject a lipid-binding protein-based complex (e.g., CER-001).

[0035] In some embodiments, the lipid-binding protein-based complex is an Apomer, a Cargomer, an HDL-based complex, or an HDL mimetic-based complex. In a specific embodiment, the lipid-binding protein-based complex is CER-001.

[0036] 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 6.1. Exemplary populations of subjects that can be treated using the compositions of the present disclosure by the methods of the present disclosure are described in Section 6.2.

[0037] In some embodiments, the methods of the present disclosure include administering a lipid - bound protein - based complex (e.g., CER - 001) to a subject in two stages. First, the lipid - bound protein - based complex (e.g., CER - 001) is administered in an initial high - intensity "induction" regimen. A lower - intensity "consolidation therapy" regimen follows the induction regimen. Alternatively, the lipid - bound protein - based complex (e.g., CER - 001) can be administered to the subject in a single stage, for example, according to an administration regimen corresponding to the dosages and frequencies of administration of the induction or consolidation therapy regimens described herein.

[0038] The induction regimens that can be used in the methods of the present disclosure are described in Section 6.3, and the consolidation therapy regimens that can be used in the methods of the present disclosure are described in Section 6.3.2. The dosing regimens of the present disclosure include administering a lipid - bound protein - based complex (e.g., CER - 001) as monotherapy or as part of a combination therapy with one or more medicaments, for example, in combination with a standard treatment for the subject's disease or condition. The combination therapies are described in Section 6.4.

[0039] 6.1. Lipid - bound protein - based complexes 6.1.1. Complexes based on HDL and HDL mimetics In one aspect, the lipid-binding protein-based complex comprises a complex based on HDL or an HDL mimetic. For example, the complex can comprise a lipoprotein complex described in U.S. Patent No. 8,206,750, International Publication No. 2012 / 109162, International Publication No. 2015 / 173633 (e.g., CER-001), or U.S. Patent Application Publication No. 2004 / 0229794, the entire contents of each of which are incorporated herein by reference. The terms “lipoprotein” and “apolipoprotein” are used interchangeably herein, and the term “lipoprotein” includes lipoprotein mimetics unless the context requires otherwise. The terms “lipid-binding protein” and “lipid-binding polypeptide” are also used interchangeably herein, and these terms do not imply an amino acid sequence of a particular length unless the context requires otherwise.

[0040] The lipoprotein complex can comprise a protein fraction (e.g., an apolipoprotein fraction) and a lipid fraction (e.g., a phospholipid fraction). Examples of the protein fraction 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 6.1.2.

[0041] Examples of the lipid fraction typically include one or more phospholipids, which can be neutral, negatively charged, positively charged, or a combination thereof. Exemplary phospholipids and other amphiphilic molecules that can be included in the lipid fraction are described in Section 6.1.3.

[0042] In certain embodiments, the lipid fraction contains at least one neutral phospholipid (e.g., sphingomyelin (SM)) and optionally one or more charged phospholipids. In lipoprotein complexes containing both neutral and charged phospholipids, the neutral and charged phospholipids can have fatty acid chains with the same or different number of carbons and the same or different degrees of saturation. In some examples, the neutral and charged phospholipids have the same acyl tail, e.g., an acyl chain of C16:0 or palmitoyl. In specific embodiments, particularly those using egg SM as the neutral lipid, the weight ratio of the apolipoprotein fraction:lipid fraction ranges from about 1:2.7 to about 1:3 (e.g., 1:2.7).

[0043] Any phospholipid that is at least partially charged at physiological pH can be used as the charged phospholipid. Non-limiting examples include phosphatidylinositol, phosphatidylserine, phosphatidylglycerol, and charged forms of phosphatidic acid, e.g., salts. In specific embodiments, the charged phospholipid is 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)], or DPPG, phosphatidylglycerol. Preferred salts include potassium and sodium salts.

[0044] In some embodiments, the lipoprotein complex used in the methods of the present disclosure is the lipoprotein complex described in U.S. Patent No. 8,206,750 or International Publication No. 2012 / 109162 (and its U.S. counterpart, U.S. Patent Application Publication No. 2012 / 0232005), the respective contents of which are incorporated herein by reference in their entirety. In certain embodiments, the protein component of the lipoprotein complex is described in Section 6.1 and preferably Section 6.1.1 of International Publication No. 2012 / 109162 (and U.S. Patent Application Publication No. 2012 / 0232005), and the lipid component is described in Section 6.2 of International Publication No. 2012 / 109162 (and U.S. Patent Application Publication No. 2012 / 0232005), which can optionally be complexed together in the amounts described in Section 6.3 of International Publication No. 2012 / 109162 (and U.S. Patent Application Publication No. 2012 / 0232005). The respective contents of these sections are incorporated herein by reference. In certain aspects, as described in Section 6.4 of International Publication No. 2012 / 109162 (and U.S. Patent Application Publication No. 2012 / 0232005), the contents of which are incorporated herein by reference, the lipoprotein complex of the present disclosure is in a population of complexes that is at least 85%, at least 90%, at least 95%, at least 97%, or at least 99% homogeneous.

[0045] In a specific embodiment, the 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.

[0046] In another specific embodiment, the 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.

[0047] In yet another specific embodiment, the lipoprotein complex that can be used in the method of the present disclosure comprises 2 to 4 ApoA-I equivalents, 2 charged phospholipid molecules, 80 lecithin molecules, and 20 SM molecules.

[0048] In yet another specific embodiment, the lipoprotein complex that can be used in the method of the present disclosure comprises 2 to 4 ApoA-I equivalents, 2 charged phospholipid molecules, 70 lecithin molecules, and 30 SM molecules.

[0049] In yet another specific embodiment, the lipoprotein complex that can be used in the method of the present disclosure comprises 2 to 4 ApoA-I equivalents, 2 charged phospholipid molecules, 60 lecithin molecules, and 40 SM molecules.

[0050] In a specific embodiment, the lipoprotein complex that can be used in the method of the present disclosure consists essentially of 2 to 4 ApoA-I equivalents, 2 charged phospholipid molecules, 50 to 80 lecithin molecules, and 20 to 50 SM molecules.

[0051] In another specific embodiment, the lipoprotein complex that can be used in the method of the present disclosure consists essentially of 2 to 4 ApoA-I equivalents, 2 charged phospholipid molecules, 50 lecithin molecules, and 50 SM molecules.

[0052] In yet another specific embodiment, the lipoprotein complex that can be used in the method of the present disclosure consists essentially of 2 to 4 ApoA-I equivalents, 2 charged phospholipid molecules, 80 lecithin molecules, and 20 SM molecules.

[0053] In yet another specific embodiment, the lipoprotein complex that can be used in the method of the present disclosure consists essentially of 2 to 4 ApoA-I equivalents, 2 charged phospholipid molecules, 70 lecithin molecules, and 30 SM molecules.

[0054] In yet another specific embodiment, the lipoprotein complex that can be used in the method of the present disclosure consists essentially of 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 60 lecithin molecules, and 40 SM molecules.

[0055] In a specific embodiment, the lipoprotein complex that can be used in the method of the present disclosure contains a lipid component comprising about 90-99.8 wt% SM and about 0.2-10 wt% charged phospholipid, 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 charged phospholipid. In another specific embodiment, the lipoprotein complex that can be used in the method of the present disclosure contains a lipid component comprising about 90-99.8 wt% lecithin and about 0.2-10 wt% charged phospholipid, 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 charged phospholipid.

[0056] In a specific embodiment, the lipoprotein complex that can be used in the method of the present disclosure contains a lipid component consisting essentially of about 90-99.8 wt% SM and about 0.2-10 wt% charged phospholipid, 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 charged phospholipid. In another specific embodiment, the lipoprotein complex that can be used in the method of the present disclosure consists essentially of about 90-99.8 wt% lecithin and about 0.2-10 wt% charged phospholipid, 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 charged phospholipid.

[0057] In yet another specific embodiment, the lipoprotein complex that can be used in the method of the present disclosure comprises about 9.8 to 90 wt% SM, about 9.8 to 90 wt% lecithin, and about 0.2 to 10 wt% charged phospholipid, for example, a lipid fraction containing a total charged phospholipid from about 0.2 to 1 wt%, 0.2 to 2 wt%, 0.2 to 3 wt%, 0.2 to 4 wt%, 0.2 to 5 wt%, 0.2 to 6 wt%, 0.2 to 7 wt%, 0.2 to 8 wt%, 0.2 to 9 wt%, 0.2 to 10 wt%.

[0058] In yet another specific embodiment, the lipoprotein complex that can be used in the method of the present disclosure comprises a lipid fraction consisting essentially of about 9.8 to 90 wt% SM, about 9.8 to 90 wt% lecithin, and about 0.2 to 10 wt% charged phospholipid, for example, a total charged phospholipid from about 0.2 to 1 wt%, 0.2 to 2 wt%, 0.2 to 3 wt%, 0.2 to 4 wt%, 0.2 to 5 wt%, 0.2 to 6 wt%, 0.2 to 7 wt%, 0.2 to 8 wt%, 0.2 to 9 wt%, 0.2 to 10 wt%.

[0059] In another specific embodiment, the lipoprotein complex that can be used in the method of the present disclosure comprises ApoA-I apolipoprotein and a lipid fraction, the lipid fraction contains sphingomyelin and about 3 wt% charged phospholipid, the molar ratio of the lipid fraction to ApoA-I apolipoprotein is about 2:1 to 200:1, and the complex is small or large discoidal particles containing 2 to 4 ApoA-I equivalents.

[0060] In another specific embodiment, the lipoprotein complex that can be used in the method of the present disclosure comprises ApoA-I apolipoprotein and a lipid fraction, the lipid fraction consists essentially of sphingomyelin and about 3 wt% charged phospholipid, the molar ratio of the lipid fraction to ApoA-I apolipoprotein is about 2:1 to 200:1, and the complex is small or large discoidal particles containing 2 to 4 ApoA-I equivalents.

[0061] Complexes based on HDL or HDL mimetics can comprise a single type of lipid-binding protein, or a mixture of two or more different lipid-binding proteins that can be from the same or different species. Optionally but preferably, the complex comprises a lipid-binding protein that is derived from the animal species being treated or has an amino acid sequence corresponding thereto, in order to avoid induction of an immune response to the treatment. Thus, for the treatment of human patients, lipid-binding proteins of human origin are preferably used. The use of peptidomimetic apolipoproteins can also reduce or avoid an immune response.

[0062] In some embodiments, the lipid component comprises two phospholipids, namely sphingomyelin (SM) and a charged phospholipid. Exemplary SMs and charged lipids are described in Section 6.1.3.1.

[0063] The lipid component comprising SM can optionally contain a small amount of additional lipid. Virtually any type of lipid can be used and is not limited thereto, including but not limited to lysophospholipids, galactocerebrosides, gangliosides, cerebrosides, glycerides, triglycerides, and cholesterol and its derivatives.

[0064] When included, such optional lipids typically comprise less than about 15 wt% of the lipid fraction, although in some examples, more optional lipids can be included. In some embodiments, the optional lipid comprises less than about 10 wt%, less than about 5 wt%, or less than about 2 wt%. In some embodiments, the lipid fraction does not contain optional lipids.

[0065] In a specific embodiment, the phospholipid fraction contains egg SM or palmitoyl SM or phytosphingomyelin and DPPG in a weight ratio (SM:charged phospholipid) in the range of 90:10 to 99:1, more preferably in the range of 95:5 to 98:2. In one embodiment, the weight ratio is 97:3.

[0066] The molar ratio of the lipid component to the protein component of the complex of the present disclosure may vary and depends, among other factors, on what the apolipoprotein containing the protein component is, what the lipid containing the lipid component is and its amount, and the desired size of the complex. Since the biological activity of apolipoproteins such as ApoA-I is thought to be mediated by amphipathic helices containing the apolipoprotein, it is convenient to use the equivalent of the ApoA-I protein to represent the apolipoprotein fraction of the lipid:apolipoprotein molar ratio. ApoA-I generally contains 6 to 10 amphipathic helices depending on the method used to calculate the helices. Other apolipoproteins can be represented in terms of ApoA-I equivalents based on the number of amphipathic helices they contain. For example, ApoA-I M which typically exists as a disulfide-bridged dimer M can be represented as 2ApoA-I equivalents because each molecule of it contains twice the number of amphipathic helices as an ApoA-I molecule. Conversely, peptide apolipoproteins containing a single amphipathic helix can be represented as 1 / 10 to 1 / 6 ApoA-I equivalents because each molecule contains 1 / 10 to 1 / 6 the number of amphipathic helices as an ApoA-I molecule. Generally, the molar ratio of lipid:ApoA-I equivalents of the lipoprotein complex (defined herein as "Ri") ranges from about 105:1 to 110:1. In some embodiments, Ri is about 108:1. The weight ratio can be obtained using an approximate MW of 650 - 800 for the phospholipid.

[0067] 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.

[0068] In some embodiments, the lipoprotein complex used in the method of the present disclosure is a charged complex comprising a protein fraction that is preferably the mature full-length ApoA-I and a lipid fraction comprising neutral lipids, sphingomyelin (SM), and charged lipids.

[0069] In a specific embodiment, the lipid component contains SM (e.g., egg SM, palmitoyl SM, phytosphingosine SM, or a combination thereof) and a charged lipid (e.g., DPPG) in a weight ratio (SM:charged lipid) in the range of 90:10 to 99:1, more preferably in the range of 95:5 to 98:2, such as 97:3.

[0070] In a specific embodiment, the ratio of the protein component to the lipid component can be in the range of about 1:2.7 to about 1:3, with 1:2.7 being preferred. This corresponds to an ApoA-I protein to lipid molar ratio in the range of approximately 1:90 to 1:140. In some embodiments, the protein to lipid molar ratio in the complex is about 1:90 to about 1:120, about 1:100 to about 1:140, or about 1:95 to about 1:125.

[0071] 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.

[0072] CER-001 as used in the literature and in the following examples refers to the complex described in Example 4 of International Publication No. WO 2012 / 109162. International Publication No. WO 2012 / 109162 refers to CER-001 as a complex having a lipoprotein weight:total lipid weight ratio of 1:2.7 and a weight:weight ratio of SM:DPPG of 97:3. Example 4 of International Publication No. WO 2012 / 109162 also describes its manufacturing method.

[0073] When used in the context of the methods of the present disclosure and / or a CER-001 dosing regimen, CER-001 refers to a lipoprotein complex in which the individual components thereof can vary by up to 20% from the CER-001 described in Example 4 of International Publication No. WO 2012 / 109162. In certain embodiments, the components of the lipoprotein complex vary by up to 10% from the CER-001 described in Example 4 of International Publication No. WO 2012 / 109162. Preferably, the components of the lipoprotein complex are those described in Example 4 of International Publication No. WO 2012 / 109162 (± variations within acceptable manufacturing tolerances). The SM in CER-001 can be natural or synthetic. In some embodiments, the SM is natural SM, such as the natural SM described in International Publication No. WO 2012 / 109162, such as chicken egg SM. In some embodiments, the SM is synthetic SM, such as the synthetic SM described in International Publication No. WO 2012 / 109162, such as synthetic palmitoyl sphingomyelin, such as that described in International Publication No. WO 2012 / 109162. Methods for synthesizing palmitoyl sphingomyelin are known in the art, such as those described in International Publication No. WO 2014 / 140787. The apolipoprotein A-I (ApoA-I), which is a lipoprotein contained in CER-001, preferably has an amino acid sequence corresponding to amino acids 25 to 267 of SEQ ID NO: 2 (previously published as SEQ ID NO: 1 in International Publication No. WO 2012 / 109162). 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 used in the dosing regimens of the present disclosure is preferably highly homogeneous, such as at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% homogeneous, as reflected by a single peak in gel permeation chromatography. See, for example, Section 6.4 of International Publication No. WO 2012 / 109162.

[0074] In a particular embodiment, the ApoA-I in CER-001 is recombinant ApoA-I produced by a mammalian host cell. The host cell can be obtained from any mammalian cell line. The polynucleotide encoding ApoA-I can be codon-optimized for expression within the recombinant host cell. Preferred host cells are mammalian host cells, including but not limited to Chinese hamster ovary cells (e.g., CHO-K1; ATCC number CCL61; CHO-S (GIBCO Life Technologies Inc., Rockville, MD, catalog number 11619012)), VERO cells, BHK (ATCC number CRL1632), BHK570 (ATCC number CRL10314), HeLa cells, COS-1 (ATCC number CRL1650), COS-7 (ATCC number CRL1651), MDCK cells, 293 cells (ATCC number CRL1573; Graham et al. J. Gen. Virol. 36:59-72, 1977), 3T3 cells, myeloma cells (especially of mouse origin), PC12 cells, and W138 cells. In certain embodiments, mammalian cells, such as CHO-S cells (Invitrogen™, Carlsbad CA), are configured for growth in serum-free media. Additional suitable cell lines are known in the art and are available from public depositories such as the American Type Culture Collection (ATCC, Manassas, VA).

[0075] In a preferred embodiment, the recombinant ApoA-I is produced by CHO cells. Recombinant ApoA-I expressed by a mammalian host cell, such as a CHO cell, may undergo post-translational modifications (e.g., glycosylation, etc.). The resulting recombinant ApoA-I may have one or more structural features (e.g., different glycosylation patterns) that are different from ApoA-I purified from human plasma.

[0076] In the case of recombinant expression of ApoA-I, a polynucleotide encoding ApoA-I is operably linked to one or more control sequences, such as a promoter or a terminator, that regulate the expression of ApoA-I in the host cell of interest. The control sequence(s) may be native or foreign to the ApoA-I coding sequence and may also be native or foreign to the host cell in which ApoA-I is expressed. Control sequences include, but are not limited to, a promoter, ribosome binding site, leader, polyadenylation sequence, propeptide sequence, signal peptide sequence, and transcription termination sequence. In some embodiments, the control sequence includes a promoter, ribosome binding site, and transcription and translation stop signals. The control sequence also includes one or more linkers for introducing specific restriction sites and may facilitate the binding of the control sequence to the coding region of the nucleotide sequence encoding ApoA-I.

[0077] The promoter driving the recombinant expression of ApoA-I can be a constitutive promoter, a regulated promoter, or an inducible promoter. Suitable promoter sequences can be obtained from genes encoding extracellular or intracellular polypeptides that are endogenous or heterologous to the host cell. Methods for isolating, identifying, and manipulating promoters of various strengths are available or can be readily applied in the art. See, for example, Nevoigt et al. (2006) Appl. Environ. Microbiol. 72:5266-5273, the disclosure of which is incorporated herein by reference in its entirety.

[0078] One or more control arrays may be derived from a viral source. For example, in certain embodiments, the promoter is derived from the polyomavirus or adenovirus major late promoter. In other embodiments, the promoter is derived from Simian Virus 40 (SV40) (Fiers et al., 1978, Nature, 273:113-120), which can be obtained as a fragment that also includes the SV40 viral origin of replication, or from cytomegalovirus, such as the simian cytomegalovirus immediate early promoter (see U.S. Patent No. 4,956,288). Other suitable promoters include those derived from the metallothionein gene (see U.S. Patent Nos. 4,579,821 and 4,601,978).

[0079] Recombinant ApoA-I expression vectors are also provided herein. The recombinant expression vector can be any vector, such as a plasmid or virus, which is operable by recombinant DNA techniques to promote the expression of heterologous ApoA-I in a recombinant host cell. The expression vector is capable of integrating into the chromosome of the recombinant host cell and contains one or more heterologous genes operably linked to one or more control sequences useful for ApoA-I production. In other embodiments, the expression vector is an extrachromosomally replicable DNA molecule, such as a linear or circular plasmid, found in a low copy number (e.g., about 1 to about 10 copies per genome equivalent) or high copy number (e.g., more than 10 copies per genome equivalent) state. In various embodiments, the expression vector contains a selectable marker, such as a gene conferring antibiotic resistance (e.g., ampicillin, kanamycin, chloramphenicol, or tetracycline resistance) to the recombinant host organism containing the vector. In a particular embodiment, the DNA constructs, vectors, and polynucleotides are suitable for ApoA-I expression in mammalian cells. Vectors for expressing ApoA-I in mammalian cells can include an origin of replication compatible with the host cell line, a promoter, and any required ribosome binding site, RNA splice site, polyadenylation site, and transcription termination sequence. In some embodiments, the origin of replication is heterologous to the host cell and is of viral origin (e.g., SV40, polyoma virus, adenovirus, VSV, BPV). In other embodiments, the origin of replication is provided by the chromosomal replication machinery of the host cell.

[0080] Methods, reagents, and tools for introducing foreign DNA into mammalian host cells are known in the art and include, but are not limited to, calcium phosphate-mediated transfection (Wigler et al., 1978, Cell 14:725; Corsaro et al., 1981, Somatic Cell Genetics 7:603; Graham et al., 1973, Virology 52:456), electroporation (Neumann et al., 1982, EMBO J. 1:841-5), DEAE-dextran-mediated transfection (Ausubel et al. (eds.), Short Protocols in Molecular Biology, 3rd Edition (John Wiley & Sons 1995)), and liposome-mediated transfection (Hawley-Nelson et al., 1993, Focus 15:73; Ciccarone et al., 1993, Focus 15:80).

[0081] In the case of high-yield production, stable expression of ApoA-I is preferred. For example, after introducing foreign DNA into a host cell, the host cell can be grown in an enriched medium for 1-2 days and then switched to a selective medium. Instead of using an expression vector containing a viral origin of replication, the host cell can be transformed with a vector containing a nucleotide sequence comprising an ApoA-I coding sequence controlled by appropriate expression control elements and a selectable marker. The selectable marker in the vector confers resistance to selection and allows the cell to stably integrate the vector into its chromosome and grow to form foci, resulting in the ability to clone and expand into a cell line. Several selection systems can be used, including, but not limited to, those containing the herpes simplex virus thymidine kinase (Wigler et al., 1977, Cell 11:223), hypoxanthine-guanine phosphoribosyl transferase (Szybalska & Szybalski, 1962, Proc. Natl. Acad. Sci. USA 48:2026), and adenine phosphoribosyl transferase (Lowy et al., 1980, Cell 22:817) genes, although they are also tk - , hgprt - , or aprt -They can each be employed intracellularly. Also, for example, dhfr that confers resistance to methotrexate (Wigler et al., 1980, Natl. Acad. Sci. USA 77:3567; O'Hare et al., 1981, Proc. Natl. Acad. Sci. USA 78:1527), gpt that confers resistance to mycophenolic acid (Mulligan & Berg, 1981, Proc. Natl. Acad. Sci. USA 78:2072), neo that confers resistance to aminoglycoside G-418 (Colberre-Garapin et al., 1981, J. Mol. Biol. 150:1), and / or hyg that confers resistance to hygromycin (Santerre et al., 1984, Gene 30:147) can be used to use anti-metabolite resistance as a criterion for selection.

[0082] Stable high-yield expression can also be achieved by using a retroviral vector that integrates into the host cell genome (see, for example, U.S. Patent Publication Nos. 2008 / 0286779 and 2004 / 0235173). Alternatively, stable high-yield expression of ApoA-I can be achieved by a gene activation method, for example, as described in International Publication No. 1994 / 012650, activating the expression of the endogenous ApoA-I gene in the genomic DNA of selected mammalian cells and requiring its amplification. By increasing the copy number of the ApoA-I gene (including the ApoA-I coding sequence and one or more control elements), it becomes possible to promote high-yield expression of ApoA-I. Preferably, the mammalian host cell in which ApoA-I is expressed has at least 2, at least 3, at least 4, or at least 5 ApoA-I gene copy indices. In a specific embodiment, the mammalian host cell in which ApoA-I is expressed has at least 6, at least 7, at least 8, at least 9, or at least 10 ApoA-I gene copy indices.

[0083] In certain embodiments, the mammalian cells are configured to produce ApoA-I in an amount of at least 0.5 g / L, at least 1 g / L, at least 1.5 g / L, at least 2 g / L, at least 2.5 g / L, at least 3 g / L, at least 3.5 g / L, and optionally, up to 4 g / L, up to 4.5 g / L, up to 5 g / L, up to 5.5 g / L, or up to 6 g / L. The mammalian host cells preferably have the ability to produce at least about 0.5, 1, 2, or 3 g / L of ApoA-I in culture and / or up to about 20 g / L of ApoA-I in culture, such as up to 4, 5, 6, 7, 8, 9, 10, 12, or 15 g / L of ApoA-I in culture.

[0084] In certain embodiments, the mammalian cells are configured to grow in serum-free medium. In these embodiments, ApoA-I is secreted from the cells. In other embodiments, ApoA-I is not secreted from the cells.

[0085] The mammalian host cells provided herein can be used to produce ApoA-I. Generally, the method includes culturing the mammalian host cells described herein under conditions in which ApoA-I is expressed. Further, the method can include recovering and optionally purifying mature ApoA-I from the supernatant of the mammalian cell culture.

[0086] The culture conditions (including culture medium, temperature, pH) can be adapted to the mammalian host cells being cultured and the selected culture mode (such as shake flask, bioreactor, roller bottle, etc.). The mammalian cells can grow in large-scale batch culture methods or in continuous or semi-continuous culture methods.

[0087] Also provided herein is a mammalian cell culture comprising a plurality of ApoA-I-producing mammalian host cells described herein. In some embodiments, the mammalian cell culture comprises at least 0.5 g / L, at least 1 g / L, at least 1.5 g / L, at least 2 g / L, at least 2.5 g / L, at least 3 g / L, at least 3.5 g / L, and optionally up to 4 g / L, up to 4.5 g / L, up to 5 g / L, up to 5.5 g / L, or up to 6 g / L of ApoA-I. The culture can be a culture of any scale ranging from about 150 mL to about 500 mL, 1 L, 10 L, 15 L, 50 L, 100 L, 200 L, 250 L, 300 L, 350 L, 400 L, 500 L, 750 L, 1000 L, 1500 L, 2000 L, 2500 L, 3000 L, 5000 L, 7500 L, 10000 L, 15000 L, 20000 L, 25000 L, 50000 L, or more. In some cases, the culture is a large-scale culture, such as 15 L, 20 L, 25 L, 30 L, 50 L, 100 L, 200 L, 300 L, 500 L, 1000 L, 5000 L, 10000 L, 15000 L, 20000 L, 25000 L, up to 50000 L, or more.

[0088] The mammalian host cells of the disclosure can grow in culture. Accordingly, the disclosure further provides for a mammalian cell culture comprising a plurality of mammalian host cells as described above. The cell culture has the following characteristics: (a) The culture (optionally, a large-scale batch culture of at least 10 liters, at least 20 liters, at least 30 liters, at least 50 liters, at least 100 liters, 300 L, 500 L, 1000 L, 5000 L, 10,000 L, 15,000 L, 20,000 L, 25,000 L, up to 50,000 L, or a continuous culture of at least 10 liters, at least 20 liters, at least 30 liters, at least 50 liters, at least 100 liters, 300 L, 500 L, 1000 L, 5000 L, or up to 10,000 L) contains or consists of a mature ApoA-I protein comprising the amino acid sequence corresponding to amino acids 25-267 of SEQ ID NO: 2 at at least about 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0 g / L or more; (b) At least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% of the protein in the culture medium is ApoA-I protein lacking the signal sequence; (c) At least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% of the protein in the culture medium is mature ApoA-I protein lacking the signal sequence and the propeptide sequence; and (d) At least 75%, at least 80%, at least 85%, at least 90%, at least 95% of the mature ApoA-I is not truncated, oxidized, or deamidated and may include one or more of the above characteristics.

[0089] CSL-111 is a complex of reconstituted human ApoA-I purified from plasma and soybean phosphatidylcholine (SBPC) (Tardif et al., 2007, JAMA 297:1675-1682).

[0090] CSL-112 is a formulation of ApoA-I that is purified from plasma, reconstituted, and forms HDL suitable for intravenous injection (Diditchenko et al., 2013, DOI 10.1161 / ATVBAHA.113.301981).

[0091] ETC-216 (also known as MDCO-216) is a lipid-depleted form of HDL containing recombinant ApoA-I ミラノ See Nicholls et al., 2011, Expert Opin Biol Ther. 11(3):387-94. doi: 10.1517 / 14712598.2011.557061.

[0092] In another embodiment, the complex that can be used in the methods of the present disclosure is CER-522. CER-522 is a lipoprotein complex comprising a combination of three phospholipids and a 22-amino acid peptide CT80522:

[0093]

Chemical formula

[0094] The phospholipid component of CER-522 consists of egg sphingomyelin, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (dipalmitoyl phosphatidylcholine, DPPC), and 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (dipalmitoyl phosphatidyl-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).

[0095] In some embodiments, the lipoprotein complex is delipidated HDL. Most HDL in plasma is cholesterol-rich. The lipids in HDL can be depleted, e.g., partially and / or selectively depleted, for example, to reduce its cholesterol content. In some embodiments, delipidated HDL may resemble the small α, preβ-1, and other preβ forms of HDL. The process of selective depletion of HDL is described in Sacks et al., 2009, J Lipid Res. 50(5): 894-907.

[0096] In certain embodiments, the lipoprotein complex comprises bioactive agent delivery particles as described in U.S. Patent Application Publication No. 2004 / 0229794.

[0097] The bioactive agent delivery particles can comprise a lipid-binding polypeptide (e.g., an apolipoprotein already described in this section or in Section 6.1.2), a lipid bilayer (e.g., one containing one or more phospholipids already described in this section or in Section 6.1.3.1), and a bioactive agent (e.g., an anti-cancer agent), wherein the interior of the lipid bilayer comprises a hydrophobic region and the bioactive agent is associated with the hydrophobic region of the lipid bilayer. In some embodiments, the bioactive agent delivery particles as described in U.S. Patent Application Publication No. 2004 / 0229794.

[0098] In some embodiments, the bioactive agent delivery particles do not comprise a hydrophilic core.

[0099] In some embodiments, the bioactive agent delivery particles are discoidal in shape (e.g., having a diameter of about 7 to about 29 nm).

[0100] The bioactive agent delivery particles include bilayer-forming lipids, such as phospholipids (e.g., those already described in this section or in Section 6.1.3.1). In some embodiments, the bioactive agent delivery particles include both bilayer-forming and non-bilayer-forming lipids. In some embodiments, the lipid bilayer of the bioactive agent delivery particles includes phospholipids. In one embodiment, examples of phospholipids incorporated into the delivery particles include dimyristoyl phosphatidylcholine (DMPC) and dimyristoyl phosphatidylglycerol (DMPG). In one embodiment, the lipid bilayer includes DMPC and DMPG in a molar ratio of 7:3.

[0101] In some embodiments, the lipid-binding polypeptide is an apolipoprotein (e.g., those already described in this section or in Section 6.1.2). The predominant interaction between a lipid-binding polypeptide, such as an apolipoprotein molecule, and the lipid bilayer is generally a hydrophobic interaction between residues on the hydrophobic face of the amphiphilic structure, such as the α-helix of the lipid-binding polypeptide, and the fatty acid acyl chains of the lipids on the outer surface of the particle periphery. Examples of bioactive agent delivery particles can include exchangeable and / or non-exchangeable apolipoproteins. In one embodiment, the lipid-binding polypeptide is ApoA-I.

[0102] In some embodiments, examples of bioactive agent delivery particles include lipid-binding polypeptide molecules, such as apolipoprotein molecules, modified to increase the stability of the particles. In one embodiment, examples of the modification include the introduction of cysteine residues to form intramolecular and / or intermolecular disulfide bonds.

[0103] In another embodiment, examples of bioactive agent delivery particles include chimeric lipid-binding polypeptide molecules, such as chimeric apolipoprotein molecules, having one or more attached functional moieties that can enhance the activity of the bioactive agent incorporated into the delivery particles or act synergistically therewith, such as one or more targeting moieties and / or one or more moieties having a desired biological activity, such as antimicrobial activity.

[0104] 6.1.2. Lipid-Binding Protein Molecules Examples of lipid-binding protein molecules that can be used in the complexes described herein include apolipoproteins such as those described in Section 6.1.2.1, and apolipoprotein mimetic peptides such as those described in Section 6.1.2.2. In some embodiments, the complex comprises a mixture of lipid-binding protein molecules. In some embodiments, the complex comprises a mixture of one or more lipid-binding protein molecules and one or more apolipoprotein mimetic peptides.

[0105] In some embodiments, the complex comprises 1 to 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, can be expressed as 2 ApoA-I equivalents because each molecule of ApoA-I contains twice the number of amphipathic helices as an ApoA-I molecule. Conversely, a peptide mimetic containing a single amphipathic helix can be expressed as 1 / 10 to 1 / 6 ApoA-I equivalents because each molecule contains 1 / 10 to 1 / 6 the number of amphipathic helices as an ApoA-I molecule. M is M because each molecule of ApoA-I contains twice the number of amphipathic helices as an ApoA-I molecule. Conversely, a peptide mimetic containing a single amphipathic helix can be expressed as 1 / 10 to 1 / 6 ApoA-I equivalents because each molecule contains 1 / 10 to 1 / 6 the number of amphipathic helices as an ApoA-I molecule.

[0106] 6.1.2.1. Apolipoproteins Suitable apolipoproteins that can be included in a complex based on a lipid-binding protein include apolipoprotein 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 foregoing. Polymorphs, isoforms, variants, and mutants of the foregoing apolipoproteins, as well as truncated forms, can also be used, the most common of which are apolipoprotein A-I Milano (ApoA-IM), apolipoprotein A-I Paris (ApoA-IP), and apolipoprotein A-I Zaragoza (ApoA-IZ). Apolipoprotein mutants containing cysteine residues are also known and can also be used (see, for example, U.S. Patent Publication No. 2003 / 0181372). The apolipoprotein can be in monomeric form, or in dimeric form which can be homodimeric or heterodimeric. For example, homodimers and heterodimers (where possible) of 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 ApoH can be used.

[0107] Apolipoproteins can have their primary sequences modified to be less susceptible to oxidation, as described, for example, in U.S. Patent Publication Nos. 2008 / 0234192 and 2013 / 0137628, and U.S. Patent Nos. 8,143,224 and 8,541,236. Apolipoproteins can contain residues corresponding to elements that facilitate their isolation, such as His tags, or other elements designed for other purposes. Preferably, the apolipoprotein in the complex is soluble in biological fluids (e.g., lymph, cerebrospinal fluid, vitreous humor, aqueous humor, blood, or blood fractions such as serum or plasma).

[0108] In some embodiments, the complex comprises a covalently bound lipid-binding protein monomer, such as the dimeric apolipoprotein A-I Milano, a variant of ApoA-I containing cysteine. Cysteine allows for the formation of disulfide bridges, which can lead to the formation of homodimers or heterodimers (e.g., ApoA-I Milano - ApoA-II).

[0109] In some embodiments, the apolipoprotein molecule comprises 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.

[0110] 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.

[0111] In some embodiments, the ApoA-I molecule is an apolipoprotein A-I Milano (ApoA-IM), apolipoprotein A-I Paris (ApoA-IP), or apolipoprotein A-I Zaragoza (ApoA-IZ) molecule.

[0112] As is well known in the art, apolipoproteins can be purified from animal sources (particularly human sources) or recombinantly produced. 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 International Publication Nos. 2008 / 104890 and 2007 / 023476. Other purification methods are also possible, such as those described in International Publication No. 2012 / 109162, the disclosure of which is hereby incorporated by reference in its entirety.

[0113] The apolipoprotein can be in prepro form, pro form, or mature form. For example, the complex can include ApoA-I (e.g., human ApoA-I), wherein the ApoA-I is prepro ApoA-I, pro ApoA-I, or mature ApoA-I. In some embodiments, the complex includes ApoA-I having at least 90% sequence identity to SEQ ID NO:1.

[0114]

Chemical Structure

[0115] In other embodiments, the complex includes ApoA-I having at least 95% sequence identity to SEQ ID NO:1. In other embodiments, the complex includes ApoA-I having at least 98% sequence identity to SEQ ID NO:1. In other embodiments, the complex includes ApoA-I having at least 99% sequence identity to SEQ ID NO:1. In other embodiments, the complex includes ApoA-I having 100% sequence identity to SEQ ID NO:1.

[0116] In some embodiments, the complex comprises ApoA-I having at least 90% sequence identity to amino acids 25 to 267 of SEQ ID NO: 2:

[0117]

Chemical formula

[0118] In some embodiments, the complex comprises 1 to 8 apolipoprotein molecules (e.g., 1 to 6, 1 to 4, 1 to 2, 2 to 8, 2 to 6, 2 to 4, 4 to 8, 4 to 6, or 6 to 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.

[0119]

[0120] ​An apolipoprotein molecule can include an apolipoprotein and one or more attached functional moieties, such as one or more CRN-001 complexes, one or more targeting moieties, moieties having a desired biological activity, affinity tags for aiding purification, and / or reporter molecules for characterization or localization studies. The attached moieties having biological activity can have an activity that can enhance and / or synergize with the biological activity of a compound incorporated into the complex of the present disclosure. For example, the moiety having biological activity can have 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 the hydrophobic surface of the complex. In another embodiment, the attached functional moiety is in contact with the hydrophobic surface of the complex. In some embodiments, the functional moiety of the chimeric apolipoprotein can be endogenous to the native 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.

[0121] In one embodiment, the chimeric apolipoprotein comprises a targeting moiety that is not endogenous to the native apolipoprotein, such as, for example, the budding yeast (S. cerevisiae) α - mating factor peptide, folic acid, transferrin, or lactoferrin. In another embodiment, the chimeric apolipoprotein comprises a moiety having a desired biological activity that enhances and / or synergizes with the activity of the compound incorporated into the complex of the present disclosure. In one embodiment, the chimeric apolipoprotein may comprise a functional moiety that is endogenous to the apolipoprotein. An example of an apolipoprotein - endogenous functional moiety is an endogenous targeting moiety substantially formed by amino acids 130 - 150 of human ApoE, including a 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 the scavenger receptor B1. In other embodiments, the functional moiety may be added synthetically or recombinantly to yield a chimeric apolipoprotein. Another example is an apolipoprotein having a prepro or pro sequence from another preproapolipoprotein (for example, one in which the prepro sequence of preproapolipoprotein A - I is replaced with the prepro sequence from preproproapolipoprotein A - II). Another example is an apolipoprotein in which a portion of the amphipathic sequence segment is replaced by another amphipathic sequence segment from another apolipoprotein.

[0122] As used herein, "chimera" refers to two or more molecules that can exist separately and are joined together to form a single molecule having all the desired functionality of its constituent molecules. The constituent molecules of a chimeric molecule can be synthetically joined together by chemical conjugation, or, if all the constituent molecules are polypeptides or analogs thereof, the polynucleotides encoding the polypeptides can be recombinantly fused together such that a single continuous polypeptide is expressed. Such chimeric molecules are referred to as fusion proteins. A "fusion protein" is a chimeric molecule in which all the constituent molecules are polypeptides and are attached (fused) to each other such that the chimeric molecule forms a continuous single chain. The various components can be attached directly 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 an apolipoprotein, or, as another example, can be added to the N-terminus or C-terminus of the sequence of an apolipoprotein. For example, a fusion protein can include an antibody light chain, an antibody fragment, a heavy chain antibody, or a single domain antibody.

[0123] In some embodiments, a chimeric apolipoprotein is prepared by chemically conjugating a functional moiety to be attached to the apolipoprotein. Means for chemically conjugating molecules are well known to those of skill in the art. Such means will vary depending on the structure of the moiety to be attached, but will be readily ascertainable by those of skill in the art. Polypeptides typically contain various 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 a linker to attach the moiety thereto. The functional moiety can be attached to a functional group on the N-terminus, C-terminus, or internal residue (i.e., one residue intermediate the N-terminus and C-terminus) of the apolipoprotein molecule. Alternatively, the apolipoprotein and / or the tagging moiety can be derivatized to expose or attach additional reactive functional groups.

[0124] In some embodiments, a fusion protein comprising a polypeptide functional portion is synthesized using a recombinant expression system. Typically, this involves creating a nucleic acid (e.g., DNA) sequence that encodes an apolipoprotein and the functional portion 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.

[0125] 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, when introduced into a suitable host cell, can be transcribed and translated into a polypeptide. The vector can include regulatory sequences such as a promoter, enhancer, or other expression control elements (e.g., polyadenylation signal). Such regulatory sequences are known to those of skill in the art (see, e.g., 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.).

[0126] In some embodiments, the apolipoprotein is modified such that when the apolipoprotein is incorporated into the complex of the present disclosure, the modification increases the stability of the complex and confers or increases the targeting ability. In one embodiment, the modification includes, for example, the introduction of cysteine residues into the apolipoprotein molecule to enable the formation of intramolecular or intermolecular disulfide bonds, such as by site-directed mutagenesis. In another embodiment, an intermolecular linkage is formed between apolipoprotein molecules using a chemical crosslinking agent to enhance the stability of the complex. The intermolecular crosslinking bond prevents or reduces the dissociation of apolipoprotein molecules from the complex and / or prevents the replacement of endogenous apolipoprotein molecules in an individual administered with the complex. In other embodiments, the apolipoprotein is modified either by chemical derivatization or site-directed mutagenesis of one or more amino acid residues to confer targeting ability or recognition thereby to cell surface receptors.

[0127] The complex can be targeted to specific cell surface receptors by engineering receptor recognition specificity into the apolipoprotein. For example, the complex can be targeted to specific cell types known to carry certain types of infectious agents by, for example, modifying the apolipoprotein to enable it to interact with receptors on the surface of the cell type to which it is targeted. For example, the complex can be targeted to macrophages by altering the apolipoprotein to confer recognition by the macrophage class A scavenger receptor (SR-A). SR-A binding ability can be conferred on the complex by modifying the apolipoprotein by site-directed mutagenesis to replace one or more positively charged amino acids with neutral or negatively charged amino acids. SR-A recognition can also be conferred by preparing chimeric apolipoproteins that contain an N- or C-terminal extension with a ligand recognized by SR-A or an amino acid sequence with a high density 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 HDL receptors, such as SR-B1.

[0128] 6.1.2.2. Apolipoprotein mimetics Peptides, peptidomimetics, 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 peptidomimetics corresponding to apolipoproteins, as well as ApoA-I, ApoA-I M, Non-limiting examples of agonists that mimic the activities of ApoA-II, ApoA-IV, and ApoE are described in U.S. Patent Nos. 6,004,925, 6,037,323, and 6,046,166 (issued to Dasseux et al.), U.S. Patent No. 5,840,688 (issued to Tso), U.S. Patent No. 6,743,778 (issued to Kohno), U.S. Patent Publication Nos. 2004 / 0266671, 2004 / 0254120, 2003 / 0171277, and 2003 / 0045460 (Fogelman), U.S. Patent Publication No. 2006 / 0069030 (Bachovchin), U.S. Patent Publication No. 2003 / 0087819 (Bielicki), U.S. Patent Publication No. 2009 / 0081293 (Murase et al.), and International Publication No. 2010 / 093918 (Dasseux et al.), the disclosures of which are incorporated herein by reference in their entireties. These peptides and peptide analogs may be composed of L-amino acids 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 well-known peptide / amide isosteres. Such apolipoprotein peptide mimetics can be synthesized or manufactured using any technique of peptide synthesis known in the art, including, for example, the techniques described in U.S. Patent Nos. 6,004,925, 6,037,323, and 6,046,166.

[0129] In some embodiments, the lipid-binding protein molecule comprises an apolipoprotein peptide mimetic molecule and optionally one or more apolipoprotein molecules, such as those described above.

[0130] In some embodiments, the apolipoprotein peptide mimetic molecule comprises an ApoA-I peptide mimetic, an ApoA-II peptide mimetic, an ApoA-IV peptide mimetic, or an ApoE peptide mimetic, or a combination thereof.

[0131] 6.1.3. Amphiphilic Molecules An amphiphilic molecule is a molecule that possesses both hydrophobic (nonpolar) and hydrophilic (polar) elements. Amphiphilic molecules that can be used in the complexes described herein include lipids (e.g., as described in Section 6.1.3.1), detergents (e.g., as described in Section 6.1.3.2), fatty acids (e.g., as described in Section 6.1.3.3), and nonpolar molecules and sterols (e.g., as described in Section 6.1.3.4) that are covalently attached to polar molecules such as, but not limited to, sugars or nucleic acids.

[0132] As a complex, it is possible to mention a single class of amphiphilic molecules (e.g., a single species of phospholipid or a mixture of phospholipids), or it can contain a combination of classes of amphiphilic molecules (e.g., phospholipids and detergents). The complex can contain one type of amphiphilic molecule or a combination of amphiphilic molecules that are configured to facilitate solubilization of lipid-binding protein molecules.

[0133] In some embodiments, the amphiphilic molecules included comprise phospholipids, detergents, fatty acids, nonpolar moieties or sterols covalently attached to sugars, or combinations thereof (e.g., selected from the types of amphiphilic molecules described above).

[0134] In some embodiments, the amphiphilic molecule comprises or consists of phospholipid molecules. In some embodiments, the phospholipid molecules include negatively charged phospholipids, neutral phospholipids, positively charged phospholipids, or combinations thereof. In some embodiments, the phospholipid molecules contribute 1 to 3 effective charges per apolipoprotein molecule in the complex. In some embodiments, the effective charge is a negative effective charge. In some embodiments, the effective charge is a positive effective 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.

[0135] In some embodiments, the amphiphilic molecule comprises a neutral phospholipid and a charged phospholipid in a weight ratio of 95:5 to 99:1.

[0136] 6.1.3.1. Lipids Complexes based on lipid-binding proteins can comprise 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, lipid molecules (e.g., phospholipid molecules) can together contribute an effective charge of 1 to 3 (e.g., 1 to 3, 1 to 2, 2 to 3, 1, 2, or 3) per lipid-binding protein molecule in the complex. In some embodiments, the effective charge is negative. In other embodiments, the effective charge is positive.

[0137] In some embodiments, the lipid comprises a phospholipid. The phospholipid can have two acyl chains that are the same or different (e.g., chains having a different number 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.

[0138] The phospholipid can have an unsaturated or saturated acyl chain with about 6 to about 24 carbon atoms (e.g., 6 to 20, 6 to 16, 6 to 12, 12 to 24, 12 to 20, 12 to 16, 16 to 24, 16 to 20, or 20 to 24). In some embodiments, the phospholipids used in the complexes 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).

[0139] Non-limiting examples of acyl chains present in commonly occurring fatty acids that can be included in phospholipids are provided in Table 1 below.

[0140] [Table 1]

[0141] Lipids that can be present in the complex of the present disclosure include, but are not limited to, small alkyl chain phospholipids, egg phosphatidylcholine, soybean 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, dimyristoyl phosphatidylglycerol, dipalmitoyl phosphatidylglycerol, distearoyl phosphatidylglycerol, dioleoyl phosphatidylglycerol, dimyristoyl phosphatidic acid, dipalmitoyl phosphatidic acid, dimyristoyl phosphatidylethanolamine, dipalmitoyl phosphatidylethanolamine, dimyristoyl phosphatidylserine, dipalmitoyl phosphatidylserine, brain phosphatidylserine, brain sphingomyelin, palmitoyl sphingomyelin, dipalmitoyl sphingomyelin, egg sphingomyelin, milk sphingomyelin, phytosphingomyelin, distearoyl sphingomyelin, dipalmitoyl phosphatidylglycerol salt, phosphatidic acid, galactosylceramide, ganglioside, cerebroside, dilauryl phosphatidylcholine, (1,3)-D-mannosyl-(1,3) diglyceride, aminophenyl glycoside, 3-cholesteryl-6'-(glycosylthio)hexyl ether glycolipid, and cholesterol and its derivatives. Lipid oxidation can be minimized using synthetic lipids such as synthetic palmitoyl sphingomyelin or N-palmitoyl-4-hydroxysphinganine-1-phosphocholine (a form of phytosphingomyelin).

[0142] In some embodiments, the lipid-binding protein-based complex comprises two types of phospholipids, namely neutral lipids such as lecithin and / or sphingomyelin (abbreviated as SM), and charged phospholipids (e.g., negatively charged phospholipids). "Neutral" phospholipids have an effective charge of approximately zero at physiological pH. In many embodiments, the neutral phospholipids are zwitterionic, although other types of net-neutral phospholipids are known and can be used. In some embodiments, the molar ratio of charged phospholipids (e.g., negatively charged phospholipids) to neutral phospholipids ranges from 1:1 to 1:3, for example, about 1:1, about 1:2, or about 1:3.

[0143] The neutral phospholipids can include, for example, one or both of lecithin and / or SM, and optionally can include other neutral phospholipids. In some embodiments, the neutral phospholipids include lecithin but not SM. In other embodiments, the neutral phospholipids include SM but not 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 in many embodiments such additional neutral phospholipids are not included.

[0144] As used herein, the expression "SM" includes sphingomyelin derived from or obtained from natural sources, as well as analogs and derivatives of naturally occurring SM that are not affected by hydrolysis by LCAT as naturally occurring SM is. SM is a phospholipid with a very similar structure to lecithin, but unlike lecithin, it does not have a glycerol backbone and thus does not have an ester linkage to which acyl chains are attached. Instead, SM has a ceramide backbone and an amide bond links the acyl chains. SM can be obtained, for example, from milk, eggs, or the brain. Analogs or derivatives 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. Synthetic SM such as synthetic palmitoyl sphingomyelin or N-palmitoyl-4-hydroxysphinganine-1-phosphocholine (phytosphingomyelin) can be used to produce a more homogeneous complex with fewer contaminants and / or oxidation products than sphingolipids of animal origin. Methods for synthesizing SM are described in U.S. Patent Publication No. 2016 / 0075634.

[0145] Sphingomyelin isolated from natural sources can be artificially enriched for one specific saturated or unsaturated acyl chain. For example, milk sphingomyelin (Avanti Phospholipid, Alabaster, Alabama) is characterized by long saturated acyl chains (i.e., acyl chains having 20 or more carbon atoms). In contrast, egg sphingomyelin is characterized by short saturated acyl chains (i.e., acyl chains having less than 20 carbon atoms). For example, only about 20% of milk sphingomyelin contains the C16:0 (16 carbons, saturated) acyl chain, while about 80% of egg sphingomyelin contains the C16:0 acyl chain. Using solvent extraction, the composition of milk sphingomyelin can be enriched to have an acyl chain composition comparable to that of egg sphingomyelin, or vice versa.

[0146] SM can be semi-synthetic to have a specific acyl chain. For example, milk sphingomyelin can first be purified from milk and then the acyl chain can be cleaved and replaced by another acyl chain, such as the C16:0 acyl chain. SM can also be fully synthesized, for example, by large-scale synthesis. See, for example, U.S. Patent No. 5,220,043 to Dong et al., issued June 15, 1993, entitled Synthesis of D-erythro-sphingomyelins, and Weis, 1999, Chem. Phys. Lipids 102 (1-2):3-12. SM can be fully synthetic, for example, as described in U.S. Patent Publication No. 2014 / 0275590.

[0147] The length and saturation level of the acyl chains, including semi-synthetic or synthetic SMs, 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, alternatively, each chain can contain a different number of carbon atoms. In some embodiments, the semi-synthetic or synthetic SMs include mixed acyl chains such that one chain is saturated and one chain is unsaturated. In such mixed acyl chain SMs, the chain lengths can be the same or different. In other embodiments, the acyl chains of the semi-synthetic or synthetic SMs 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 including the semi-synthetic or synthetic SMs are identical. In specific embodiments, the chains correspond to acyl chains of naturally occurring fatty acids such as oleic acid, palmitic acid, or stearic acid. In another embodiment, SMs having saturated or unsaturated functionalized chains are used. In another specific embodiment, both acyl chains are saturated and contain 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.

[0148] In some embodiments, the SM is palmitoyl SM, such as synthetic palmitoyl SM having a C16:0 acyl chain, or egg SM containing palmitoyl SM as a main component.

[0149] In specific embodiments, functionalized SMs such as phytosphingomyelin are used.

[0150] Lecithin can be derived from or isolated from natural sources, or it can be obtained synthetically. Examples of suitable lecithins isolated from natural sources include, but are not limited to, egg phosphatidylcholine and soybean phosphatidylcholine. Further non-limiting examples of suitable lecithins 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-palmitoyl phosphatidylcholine, dioleoyl phosphatidylcholine, and ether derivatives or analogs thereof.

[0151] Lecithin derived from or isolated from natural sources can be enriched to contain a specified acyl chain. In embodiments using semi-synthetic or synthetic lecithin, as described above in connection with SM, the acyl chain can be selectively varied as to what it is. In some embodiments of the complexes described herein, both acyl chains on the lecithin are the same. In some embodiments of complexes containing both SM and lecithin, all of the acyl chains of SM and lecithin are the same. In specific embodiments, the acyl chain corresponds to the acyl chain of myristic acid, palmitic acid, oleic acid, or stearic acid.

[0152] The complex 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, "negatively charged phospholipid" refers to 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 different 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 salt or potassium salt). 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.

[0153] The charged phospholipids can be obtained from natural sources or prepared by chemical synthesis. In embodiments using synthetic charged phospholipids, as described above in relation to SM, the acyl chain can be selectively varied as to what it is. In some embodiments of the complexes of the present disclosure, both acyl chains on the 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 complex includes a charged phospholipid and / or SM having all C16:0 or C16:1 acyl chains. In a specific embodiment, the fatty acid moiety of SM is mainly C16:1 palmitoyl. In a specific embodiment, the acyl chains of the charged phospholipid, lecithin and / or SM correspond to the acyl chains of palmitic acid. In yet another specific embodiment, the acyl chains of the charged phospholipid, lecithin and / or SM correspond to the acyl chains of oleic acid.

[0154] 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-aminopropyl)amino]butylcarboxamide)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-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-3-dimethylammonium-propane 1,2-dimyristoyl-3-dimethylammonium-propane, 1,2-dipalmitoyl-3-dimethylammonium-propane, N-(4-carboxybenzyl)-N,N-dimethyl-2,3-bis(oleyloxy)propane-1-aminium, 1,2-dioleoyl-3-trimethylammonium-propane, 1,2-dioleoyl-3-trimethylammonium-propane, 1,2-stearoyl-3-trimethylammonium-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)-1 propanaminium methyl sulfate, and salts thereof (e.g., chloride or bromide salts).

[0155] The lipid to be used is preferably at least 95% pure and / or has a reduced level of oxidizing agents (including but not limited to peroxides, etc.). Lipids obtained from natural sources preferably have fewer polyunsaturated fatty acid moieties and / or fatty acid moieties that are not sensitive to oxidation. The level of oxidation in the sample can be determined using an iodometric method that provides a peroxide value, which is expressed as the milliequivalent number of isolated iodine per kg of sample and 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, F.W. 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 level, is low, e.g., 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.

[0156] In some embodiments, the complex can include a small amount of additional lipid. Virtually any type of lipid can 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 complexes of the present disclosure can contain cholesterol or a cholesterol derivative, such as a cholesterol ester. The cholesterol derivative can also be a substituted cholesterol or a substituted cholesterol ester. The complexes of the present disclosure can 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 include cholesterol and / or its derivatives (cholesterol esters or oxidized cholesterol esters).

[0157] 6.1.3.2. Detergent The complex can contain one or more detergents. The detergents can 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 nonionic 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.

[0158] 6.1.3.3. Fatty Acids The complex can contain one or more fatty acids. The one or more fatty acids can include a single-chain fatty acid having an aliphatic tail of 5 or fewer carbons (e.g., butyric acid, isobutyric acid, valeric acid, or isovaleric acid), a medium-chain fatty acid having an aliphatic tail of 6 to 12 carbons (e.g., caproic acid, caprylic acid, capric acid, or lauric acid), a long-chain fatty acid having an aliphatic tail of 13 to 21 carbons (e.g., myristic acid, palmitic acid, stearic acid, or arachidic acid), a very-long-chain fatty acid having an aliphatic tail of 22 or more carbons (e.g., behenic acid, lignoceric acid, or cerotic acid), or a combination thereof. The one or more fatty acids can 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, linolelaidic acid, α-linolenic acid, arachidonic acid, eicosapentaenoic acid, erucic acid, or docosahexaenoic acid), or a combination thereof. The unsaturated fatty acid can be a cis or trans fatty acid. In some embodiments, the unsaturated fatty acid used in the complexes of the present disclosure is a cis fatty acid.

[0159] 6.1.3.4. Sugar and Attached Nonpolar Molecules and Sterols The complex can contain one or more amphiphilic molecules including a nonpolar molecule or moiety (e.g., a hydrocarbon chain, acyl, or diacyl chain) or a sterol (e.g., cholesterol) attached to 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 sugar or a substituted sugar. Exemplary amphiphilic molecules containing a sugar and an attached nonpolar molecule include dodecan-2-yl O-β-D-maltoside, tridecan-3-yl O-β-D-maltoside, tridecan-2-yl O-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.

[0160] In some embodiments, the nonpolar moiety is an acyl or diacyl chain.

[0161] In some embodiments, the sugar is a modified sugar or a substituted sugar.

[0162] 6.1.4. Formulations Complexes based on lipid-binding proteins can be formulated for the intended route of administration 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).

[0163] CER-001, intended for administration by infusion, can be formulated in a phosphate buffer containing sucrose and mannitol excipients, as described, for example, in International Publication No. WO 2012 / 109162.

[0164] 6.2. Target Population Subjects that can be treated according to the methods described herein are preferably mammals, most preferably humans.

[0165] In some embodiments, the subject has a condition that includes inflammation (e.g., acute inflammation and / or hyperinflammation).

[0166] In some embodiments, the subject has or is at risk of having HLH. In some embodiments, the subject has HLH. In other embodiments, the subject is at risk of having HLH. HLH can be, for example, familial HLH, or HLH subsequent to a malignant disease (e.g., acute leukemia or lymphoma) or a non-malignant disease (e.g., an autoimmune disease, such as rheumatoid arthritis, or an infectious disease, such as a viral or bacterial infection). In some embodiments, HLH is virus-induced HLH caused by, for example, dengue infection, herpes simplex infection, or Epstein-Barr infection.

[0167] In some embodiments, the subject has or is at risk of having dengue hemorrhagic fever or dengue shock syndrome, e.g., the subject has a dengue infection (e.g., the subject has dengue). Dengue, dengue hemorrhagic fever, and dengue shock syndrome are described in Dengue haemorrhagic fever : diagnosis, treatment, prevention and control, 2 nd Edition, World Health Organization (1997) (the contents of which are incorporated herein by reference in their entirety). Subjects with dengue are at risk of progressing to more severe dengue hemorrhagic fever and even more severe dengue shock syndrome.

[0168] In some embodiments, the subject has a herpes simplex infection.

[0169] In some embodiments, the subject has a SOFA score of 1 to 4, such as a score of 1, 2, 3, or 4, prior to treatment with a lipid-binding protein-based complex (see Vincent et al. 1996, Intensive Care Med, 22:707-710).

[0170] In some embodiments, the subject has or is at risk of acute kidney injury (AKI) due to, for example, a viral infection (such as a dengue infection).

[0171] In some aspects, the subject may have or be at risk of CRS and / or may need to reduce the serum levels of one or more inflammatory markers (such as IL-6). In some embodiments, the subject has CRS. In some embodiments, the subject has CRS following an infection, such as a viral infection, such as a dengue infection. In still other embodiments, the subject is at risk of CRS due to, for example, an infection, such as dengue.

[0172] In another aspect, the subject is a subject in need of a decrease in the serum level of one or more inflammatory markers, e.g., a subject having an elevated level of one or more inflammatory markers as 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 α (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, e.g., 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 α (TNFα).

[0173] 6.3. Dosage Regimen While single doses may be used in some embodiments, the methods of the disclosure generally require multiple administrations, e.g., 2 to 10 individual doses, of a lipid-binding protein-based complex (e.g., CER-001). In some embodiments, the dosing regimen may include two or more, three or more, or four or more doses, e.g., 5, 6, 7, 8, 9, 10, 11, 12, or more than 12 doses, of a lipid-binding protein-based complex (e.g., CER-001). In some embodiments, the dosing regimen includes or consists of a single dose. In some embodiments, the dosing regimen includes or consists of two individual doses. In some embodiments, the dosing regimen includes or consists of three individual doses. In some embodiments, the dosing regimen includes or consists of four individual doses.

[0174] In some embodiments, the lipid-binding protein-based complexes are administered according to the induction and optionally the consolidation therapy regimens described in Sections 6.3.1 and 6.3.2, respectively. In some embodiments, the lipid-binding protein-based complexes can be administered in a single step, for example, according to the administration regimens described in this section. In some embodiments, the subject does not undergo treatment with a lipid-binding protein-based complex according to a maintenance regimen, for example, a regimen that includes administration of a lipid-binding protein-based complex for an extended period (e.g., for more than one month).

[0175] The administration regimens of the lipid-binding protein-based complexes (e.g., CER-001) of the present disclosure can last up to one week, one week, or longer than one week (e.g., two weeks).

[0176] For example, the administration regimen of a lipid-binding protein-based complex (e.g., CER-001) can include administering: - Five doses of CER-001 over one week, - Six doses of CER-001 over one week, - Seven doses of CER-001 over one week, - Ten doses of CER-001 over two weeks, - Twelve doses of CER-001 over two weeks, - Fourteen doses of CER-001 over two weeks.

[0177] In one embodiment, the method of the present disclosure includes administering seven doses of CER-001 over one week, for example, on days 1, 2, 3, 4, 5, 6, and 7.

[0178] In some embodiments of the methods of the present disclosure, multiple doses of a lipid-binding protein-based complex (e.g., CER-001) are administered at intervals of 1 day or less. For example, in some embodiments, two or more individual doses are administered at approximately 12-hour intervals. In some embodiments, two individual doses are administered at approximately 12-hour intervals. In other embodiments, three individual doses are administered at approximately 12-hour intervals. In other embodiments, two individual doses are administered at approximately 12-hour intervals, and the third individual dose is administered approximately 1 day later. In other embodiments, three individual doses are administered at approximately 12-hour intervals, and the fourth individual dose is administered approximately 1 day later.

[0179] In some embodiments of the methods of the present disclosure, a lipid-binding protein-based complex (e.g., CER-001) is administered to a subject at 0 and 12 hours, for example, at a dose of 10 mg / kg or 15 mg / kg (e.g., over 0.5 - 1 hour). In some embodiments of the methods of the present disclosure, a lipid-binding protein-based complex (e.g., CER-001) is administered to a subject at 0 hours, and at 12, 24, and 48 hours, for example, at a dose of 10 mg / kg or 15 mg / kg (e.g., over 0.5 - 1 hour).

[0180] In some embodiments of the methods of the present disclosure, a lipid-binding protein-based complex (e.g., CER-001) is administered daily, for example, for at least 5 days, at least 6 days, at least 7 days, or longer than 7 days (e.g., daily for up to 1 week or up to 2 weeks). In other embodiments, a lipid-binding protein-based complex (e.g., CER-001) is administered at a lower frequency, for example, every other day, twice a week, three times a week, or once a week.

[0181] In practice, an administration window can be provided to account for minor variations in the dosing schedule, for example, for multiple dosing per week. For example, a window of ±2 days or ±1 day around the dosing day can be used.

[0182] Complexes based on lipid-binding proteins (e.g., CER-001) can be administered in the methods of the present disclosure for a pre-determined period of time, e.g., for one week. Alternatively, administration of a lipid-binding protein-based complex (e.g., CER-001) can continue until one or more symptoms of a condition (e.g., HLH or dengue shock syndrome) are alleviated, or until the serum levels of one or more inflammatory markers decrease, e.g., decrease to normal levels or decrease relative to a baseline value in the subject, e.g., a baseline value measured prior to initiating treatment with a lipid-binding protein-based complex (e.g., CER-001). Reference or “normal” levels for various inflammatory markers are known in the art. For example, the Mayo Clinic Laboratories test catalog (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 (male), 11 - 307 mcg / L (female), IL-8 < 57.8 pg / mL, TNF-α < 5.6 pg / mL.

[0183] The methods of the present disclosure generally involve administering a high dose of a lipid-binding protein-based complex (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, or 10 individual doses), and can be administered, for example, over one day or multiple days (e.g., a one-day period, a two-day period, a three-day period, 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). In some embodiments, the individual doses of the high dose are administered daily, twice a day, or at 2 - 3 day intervals.

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

[0185] In some embodiments, the high dose is an amount effective to reduce the 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, and MCP1 TNF-α. In some embodiments, the serum levels of one or more inflammatory markers decrease from elevated ranges to normal ranges and / or decrease by at least 20%, at least 40%, or at least 60%.

[0186] The dosage of a lipid-binding protein-based complex (e.g., CER-001) to be administered to a subject (e.g., an individual dosage that forms a high dosage when combined with one or more other individual dosages) is, in some embodiments, 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 foregoing 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 "on a protein weight basis" means that the dosage of the lipid-binding protein-based complex (e.g., CER-001) to be administered to a subject is calculated based on the amount of ApoA-I in the lipid-binding protein-based complex (e.g., CER-001) to be administered and the body weight of the subject. For example, a subject with a body weight of 70 kg receiving a dosage of CER-001 of 20 mg / kg would receive an amount of CER-001 that provides 1400 mg of ApoA-I (70 kg × 20 mg / kg).

[0187] In yet other embodiments, the lipid-binding protein-based complex (e.g., CER-001) can be administered on a unit dosage basis. The unit dosage used in the methods of the present disclosure can vary, in some embodiments, from 300 mg to 4000 mg (e.g., 600 mg to 4000 mg) / administration (on a protein weight basis).

[0188] In certain embodiments, the dosage of the lipid-binding protein-based complex (e.g., CER-001) is 600 mg to 3000 mg, 800 mg to 3000 mg, 1000 mg to 2400 mg, or 1000 mg to 2000 mg / administration (on a protein weight basis).

[0189] In one aspect, a high dose of a lipid - bound protein - based complex (e.g., CER - 001), for example, the collection of multiple individual doses, is from 600 mg to 40 g (protein weight basis). In certain embodiments, the high dose is from 3 g to 35 g or from 5 g to 30 g (protein weight basis).

[0190] The lipid - bound protein - based complex (e.g., CER - 001) is preferably administered as an IV infusion. For example, the stock solution of CER - 001 can be diluted to a total volume of 125 - 250 ml with a standard saline such as physiological saline (0.9% NaCl). In some embodiments, subjects weighing less than 80 kg will have a total volume of 125 ml, while subjects weighing at least 80 kg will have a total volume of 250 ml. In some embodiments, the dose of CER - 001 is administered at a total volume of 250 ml. The lipid - bound protein - based complex (e.g., CER - 001) can be administered over a period ranging from 1 hour to 24 hours. Depending on the requirements of the subject, the administration can be by a slow infusion over a period longer than 1 hour (e.g., up to 2 hours or up to 24 hours), by a rapid infusion of less than 1 hour, or by a single bolus injection. In one embodiment, the lipid - bound protein - based complex (e.g., CER - 001) is administered over a 1 - hour period, for example, using an infusion pump at a fixed rate of 125 ml / h or 250 ml / h. In one embodiment, the dose of the lipid - bound protein - based complex (e.g., CER - 001) is administered as an infusion over a 24 - hour period.

[0191] 6.3.1. Induction Regimen In one embodiment, an induction regimen suitable for use in the methods of the present disclosure involves administering multiple doses of a lipid - bound protein - based complex (e.g., CER - 001) over multiple consecutive days, for example, over 3 consecutive days.

[0192] In some embodiments, an induction regimen suitable for use in the methods of the present disclosure involves administration of a lipid-bound protein-based complex (e.g., CER-001) twice daily, e.g., twice-daily administration for several consecutive days. Twice-daily administration can include, for example, two doses at approximately 12-hour intervals or a morning dose and an evening dose (which can be at intervals longer or shorter than 12 hours).

[0193] In one embodiment, the induction regimen includes a lipid-bound protein-based complex (e.g., CER-001) at a dose of twice daily for 3 consecutive days.

[0194] The therapeutic dose of the lipid-bound protein-based complex (e.g., CER-001) administered by infusion in the induction regimen can range from 4 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 foregoing 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 the lipid-bound protein-based complex (e.g., CER-001) used in the induction regimen is 5 mg / kg. In some embodiments, the dose of the lipid-bound protein-based complex (e.g., CER-001) used in the induction regimen is 10 mg / kg. In some embodiments, the dose of the lipid-bound protein-based complex (e.g., CER-001) used in the induction regimen is 15 mg / kg. In some embodiments, the dose of the lipid-bound protein-based complex (e.g., CER-001) used in the induction regimen is 20 mg / kg. In some embodiments, the induction regimen includes a lipid-bound protein-based complex (e.g., CER-001) at a dose of 5 mg / kg, 10 mg / kg, 15 mg / kg, or 20 mg / kg for 6 doses over 3 days.

[0195] In yet other embodiments, lipid-bound protein-based complexes (such as CER-001) can be administered on a per unit dosage basis. The unit dosage used in the induction phase may vary for infusion administration from 300 mg to 4000 mg (such as 300 mg to 3000 mg) (protein weight basis).

[0196] In certain embodiments, the dosage of the lipid-bound protein-based complex (such as 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) for infusion administration.

[0197] 6.3.2. Consolidation Therapy Regimen Consolidation therapy regimens suitable for use in the methods of the present disclosure involve administering one or more doses of a lipid-bound protein-based complex (such as CER-001) following the induction regimen.

[0198] In one embodiment, the consolidation therapy regimen includes administering two doses of a lipid-bound protein-based complex (such as CER-001). For example, the two doses can be administered at approximately 12-hour intervals, or as a morning dose and an evening dose (which can be at intervals longer or shorter than 12 hours).

[0199] The dosage of a lipid-binding protein-based complex (e.g., CER-001) in a consolidation therapy regimen can, in some embodiments, be administered on the 6th day of a dosing regimen starting from the induction regimen on day 1. The dosage of a lipid-binding protein-based complex (e.g., CER-001) in a consolidation therapy regimen can, in some embodiments, be administered on the 4th day of a dosing regimen starting from the induction regimen on day 1. The dosage of a lipid-binding protein-based complex (e.g., CER-001) in a consolidation therapy regimen can, in some embodiments, be administered on the 5th day of a dosing regimen starting from the induction regimen on day 1. The dosage of a lipid-binding protein-based complex (e.g., CER-001) in a consolidation therapy regimen can, in some embodiments, be administered on the 7th day of a dosing regimen starting from the induction regimen on day 1.

[0200] In a consolidation therapy regimen, the therapeutic dose of a lipid-binding protein-based complex (e.g., CER-001) administered by infusion can range from 4 mg / kg to 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 foregoing values, e.g., 5 - 15 mg / kg, 10 - 20 mg / kg, or 15 - 25 mg / kg). In some embodiments, the dose of the lipid-binding protein-based complex (e.g., CER-001) used in a consolidation therapy regimen is 5 mg / kg. In some embodiments, the dose of the lipid-binding protein-based complex (e.g., CER-001) used in a consolidation therapy regimen is 10 mg / kg. In some embodiments, the dose of the lipid-binding protein-based complex (e.g., CER-001) in a consolidation therapy regimen is 15 mg / kg. In some embodiments, the dose of the lipid-binding protein-based complex (e.g., CER-001) used in a consolidation therapy regimen is 20 mg / kg. In some embodiments, the consolidation therapy regimen comprises a lipid-binding protein-based complex (e.g., CER-001) at a dose of 5 mg / kg, 10 mg / kg, 15 mg / kg, or 20 mg / kg, administered as two doses in one day.

[0201] In yet other embodiments, the lipid-binding protein-based complex (e.g., CER-001) can be administered on a unit dosage basis. The unit dosage used in the consolidation therapy phase can vary from 300 mg to 4000 mg (e.g., 300 mg to 3000 mg) (protein weight basis) / infusion administration.

[0202] In certain embodiments, the dosage of the lipid-binding protein-based complex (e.g., CER-001) used during the consolidation therapy phase is from 300 mg to 1500 mg, 400 mg to 1500 mg, 500 mg to 1200 mg, or 500 mg to 1000 mg (protein weight basis) / infusion administration.

[0203] Complexes based on lipid-binding proteins (e.g., CER-001) can be administered during the consolidation therapy phase in the same manner as described in Section 6.3, for example, as an IV infusion over a period of 1 hour.

[0204] 6.4. Combination Therapy Complexes based on lipid-binding proteins (e.g., CER-001) can be administered to the subjects described herein as part of a monotherapy or combination therapy regimen. For example, the combination therapy can include a complex based on a lipid-binding protein (e.g., CER-001) in combination with standard treatment for sepsis and / or AKI. 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.

[0205] In some embodiments, the subject is treated using a lipid-binding protein-based complex (e.g., CER-001) in combination with fluid replacement therapy. In some embodiments, the subject is treated using a lipid-binding protein-based complex (e.g., CER-001) in combination with an antimicrobial. In some embodiments, the subject is treated using a lipid-binding protein-based complex (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 using a lipid-binding protein-based complex (e.g., CER-001) in combination with an antiviral agent. In some embodiments, the subject is treated using a lipid-binding protein-based complex (e.g., CER-001) in combination with an agent that raises blood pressure (e.g., norepinephrine or epinephrine). In some embodiments, the subject is treated using a lipid-binding protein-based complex (e.g., CER-001) in combination with an immunosuppressant, such as tacrolimus or everolimus.

[0206] The combination therapy regimen can, in some embodiments, include one or more other agents for treating CRS, such as one or more anti-IL-6 agents and / or corticosteroids (e.g., methylprednisolone and / or dexamethasone). Exemplary anti-IL6 agents include tocilizumab, siltuximab, olaratumab, elsilimomab, BMS-945429, sirukumab, lebrikizumab, and CPSI-2364. In some embodiments, the lipid-binding protein-based complex (e.g., CER-001) is administered in combination with tocilizumab.

[0207] In certain embodiments, an antihistamine (e.g., diphenhydramine, cetirizine, fexofenadine, or loratadine) can be administered prior to administration of a lipid-binding protein-based complex (e.g., CER-001). The antihistamine can reduce the likelihood of an allergic reaction.

Example

[0208] 7. Example [Example 1] 7.1. CER-001 Treatment for COVID-19 The SARS-CoV-2 virus can promote life-threatening hyperinflammatory states in at-risk patients. Remodeling of the lipid profile, including a dramatic decrease in serum levels of apolipoprotein A-I (ApoA-I), is characteristic of severe COVID-19. ApoA-I can reduce lung inflammation, regulate innate and adaptive immunity, and prevent endothelial dysfunction and blood coagulation. In this example, a compassionate access trial is described for four subjects with progressive COVID-19 cytokine storm despite standard treatment. To raise ApoA-I to normal levels, the subjects received 2-4 infusions of CER-001 (10 mg / kg each). The injections showed good tolerance and no serious adverse events were observed. Three patients improved rapidly and were discharged 3-4 days after the CER-001 infusion. In the fourth patient, who received CER-001 administration while on mechanical ventilation, a temporary improvement was seen, but then deterioration related to bacterial pneumonia occurred. This trial provides initial safety data and proof-of-concept data for treating patients with a virus-induced cytokine storm while using a lipid-binding protein-based complex, such as CER-001.

[0209] 7.1.1. Materials and Methods 7.1.1.1. Medical History Subject 1 was a 52-year-old male with a history of IgA vasculitis, diabetes, and ischemic heart disease, who had received a kidney transplant in 2018. He had received three doses of the mRNA COVID-19 vaccine but developed only weak anti-SARS-CoV-2 immunity (anti-spike antibody 15.5 BAU / mL). The subject developed symptoms of COVID-19 (fever, diarrhea, and dyspnea) and was admitted to the transplant ward 8 days later. Oxygen saturation in room air was 92%, and oxygen supplementation (1 L / min) was initiated. Chest CT scan revealed bilateral interstitial lung disease consistent with COVID-19 (25% of the parenchymal infiltration), and nasopharyngeal PCR detected the SARS-CoV-2 Delta strain (variant of concern (VOC)). Blood tests showed a hyperinflammatory state (ferritin 5,037 μg / L, C-reactive protein 34 mg / L), abnormal liver tests (AST and ALT were 2.5 and 3.5 times the upper limit of normal (ULN), respectively), and thrombocytopenia. Tacrolimus was administered, mycophenolate mofetil was discontinued, and dexamethasone (6 mg / day) was introduced along with antibiotics. Blood tests on the second day revealed pancytopenia and progression of the hyperinflammatory state (ferritin 6,870 μg / L, C-reactive protein 55 mg / L). The subject received one infusion of the monoclonal antibody anti-IL-6R antibody tocilizumab (8 mg / kg, i.v.) and one infusion of the neutralizing monoclonal anti-SARS-CoV-2 antibody (casirivimab / imdevimab). On the fourth day, the subject's ferritin increased to 19,219 μg / L, the subject's AST and ALT increased to 17 and 14 times the ULN values, respectively, and the subject's arterial lactate level was 2.7 mmol / L. Bone marrow aspiration revealed findings of hemophagocytosis. Blood PCR for SARS-CoV-2 was weakly positive. The worsening of hypoxemia required an increase in the oxygen supply (4 L / min; PaO2 62 mmHg), and CT scan revealed progressive lung lesions typical of COVID-19 (50% of the parenchyma). Despite increasing dexamethasone to 10 mg / day, on the sixth day, serum triglycerides and ferritin increased to 3.2 mmol / L and 27,394 μg / L, respectively.

[0210] Subject 2 was a 38-year-old woman with a history of systemic lupus erythematosus (SLE) and obesity, who had received a kidney transplant in 2011. She had received three doses of the mRNA COVID-19 vaccine but did not acquire anti-SARS-CoV-2 immunity. The subject developed symptoms of COVID-19 (cough, chills, diarrhea, fever) and was admitted to the transplant ward 10 days later. The SARS-CoV-2 VOC Omicron strain was identified by nasopharyngeal PCR testing. At admission, SaO2 was 94% while receiving oxygen inhalation at 9 L / min while wearing a face mask. Chest CT scan revealed typical lesions of COVID-19 (infiltration range 50%). Blood tests revealed hepatitis with cytolysis and cholestasis (7- to 10-fold the ULN values respectively), acute kidney injury (KDIGO stage 1), and a hyperinflammatory state (ferritin 2,000 μg / L, C-reactive protein 107 mg / L). High-flow oxygen supplementation, awake proning, dexamethasone (10 mg / day), tocilizumab (8 mg / kg as a single dose), and antibiotics were initiated. Everolimus was discontinued and tacrolimus was administered. On the fourth day, despite maximal treatment, high-flow oxygen supplementation was still required and the subject's hyperinflammatory state worsened (ferritin 2,800 μg / L).

[0211] Subject 3 was a 47-year-old woman with a history of diabetes, adrenal Cushing's syndrome, hypertension, and end-stage renal disease requiring chronic renal replacement therapy since 2020. The subject had not received anti-SARS-CoV-2 vaccination and had no anti-SARS-CoV-2 immunity at the time of admission. She developed symptoms of COVID-19 (cough, dyspnea, abdominal pain, fever) and was admitted to the hospital 4 days later. SARS-CoV-2 VOC Omicron strain was identified by nasopharyngeal PCR. Chest CT scan revealed mild to moderate lung lesions (10 - 25%) typical of COVID-19. Oxygen supplementation was not required. Blood tests revealed a hyperinflammatory syndrome (ferritin 4,350 μg / L, C-reactive protein 55 mg / L), moderate elevation of AST and ALT (2 times and 1.5 times the ULN values respectively), as well as mild thrombocytopenia and anemia. Dexamethasone (6 mg / day) was introduced. On the 3rd day, hyperferritinemia (ferritin 4,142 μg / L) and abnormal liver test items persisted, and the subject developed encephalopathy and was admitted to the intensive care unit.

[0212] Subject 4 was a 59-year-old man with a history of hepatitis B, liver transplantation in 2006, HHV8-negative Kaposi's sarcoma (complete remission), and end-stage renal disease requiring chronic renal replacement therapy since 2020. The subject had received three doses of mRNA COVID-19 vaccine but did not express anti-SARS-CoV-2 antibodies. Due to SARS-CoV-2 exposure within the family, a nasopharyngeal PCR test was performed and the VOC Omicron strain was identified. The subject developed symptoms of COVID-19 (malaise) but initially had no respiratory symptoms. Chest CT scan revealed mild to moderate lung lesions (10 - 25%) typical of COVID-19. Two days later, he developed dyspnea, cough, and fever. At the time of admission, PaO2 was 54 mmHg in room air, respiratory rate was 30 breaths / min, and body temperature was 38.5°C. Blood tests revealed hyperferritinemia (ferritin 1,223 μg / L) and leukopenia (1,080 cells / mm 3) became apparent. CT scan revealed progression of the lung lesion (25 - 50%). Oxygen supplementation, dexamethasone (10 mg / day), tocilizumab (administered once at 8 mg / kg), antibiotics, and fresh frozen plasma obtained from convalescent patients were administered. Mycophenolate mofetil was discontinued and tacrolimus was administered. On the 5th day, acute respiratory failure developed and mechanical ventilation with oral endotracheal intubation and neuromuscular blockade was required. Blood tests revealed a hyperinflammatory state (ferritin 4,535 μg / L) accompanied by an increase in AST and ALT (3 times the ULN value). At that time, the culture test of the bronchoalveolar lavage fluid was negative, suggesting only severe COVID-19. The PaO2 / FiO2 ratio was 150 - 180. Antibiotics were administered.

[0213] 7.1.1.2. Administration method CER-001 was intravenously administered to Subject 1 at a dose of 10 mg / kg at 0 hour and 12 hours over a period of 0.5 - 1 hour. CER-001 was intravenously administered to Subjects 2 - 4 at a dose of 10 mg / kg at 0, 12, 24, and 48 hours over a period of 0.5 - 1 hour. Prior to each administration of CER-001, prophylactic administration of an antihistamine drug using hydroxyzine (50 mg i.v.) preceded. Dexamethasone was also administered to all subjects.

[0214] 7.1.2. Results and discussion 7.1.2.1. General safety Subjects 1, 2, and 3 did not exhibit any serious adverse events. Subject 4 exhibited two episodes of ventilator-associated pneumonia (VAP; Klebsiella pneumoniae and Aspergillus fumigatus + mucormycosis), and one episode of bacteremia (Staphylococcus haemolyticus).

[0215] 7.1.2.2. Biological efficacy: Lipid profile When CER-001 was first administered, in all 4 subjects, the serum levels of ApoA-I (in the range of 0.74 - 0.79 mg / L, normal value > 1.1 g / L) (Figs. 2A - 2D), and HDL (in the range of 0.26 - 0.35 g / L, normal value > 0.45 g / L) (Figs. 3A - 3D) were very low, while the serum level of triglyceride was high (in the range of 2.16 - 3.4 g / L, normal value < 1.5 g / L). After treatment with CER-001, on the 2nd day, ApoA-I and HDL levels normalized in all subjects, but remained low within the normal value range in the most inflammatory subject. In subject 4 who developed ventilator-associated pneumonia 3 days after the start of CER-001, ApoA-I then decreased below the normal value.

[0216] 7.1.2.3. Kinetics of inflammation At baseline, IL-1β was normal in all individuals, but for IL-6, it increased in 3 subjects who had received tocilizumab previously, and was normal in the 4th subject (3.3 - 1,295 pg / mL), while TNF-α increased moderately (9.7 - 42.1 pg / mL). IL-8 was the only inflammatory cytokine that increased in all cases (> 10 pg / mL; 14.8 - 64.5 pg / mL). After administration of CER-001, IL-8 normalized in subjects 1, 2, and 3. In subject 4, IL-8 decreased rapidly after injection but increased again at the onset of ventilator-associated pneumonia. The serum ferritin level decreased from 6,616 ± 8,696 μg / L to 1,712 ± 815 μg / L 6 days after the start of CER-001. In 3 out of 4 subjects, the analysis of C-reactive protein was excluded because an anti-IL-6R antibody had been administered before CER-001. The body temperature remained below 37.5°C in all subjects.

[0217] 7.1.2.4. Clinical outcome After administration of CER-001, the clinical conditions of Subjects 1, 2, and 3 improved rapidly, and they were able to be discharged 3 to 4 days after the infusion of CER-001 (Figures 1A to 1D). In Subjects 1 and 2, oxygen supplementation was discontinued 2 and 3 days after administration, respectively. In Subject 3, the confused state dissipated within 2 days. In these 3 subjects, inflammatory parameters, liver test values, and blood cell counts improved by the time of discharge. Subject 4 had been receiving mechanical ventilation for 3 days at the time of CER-001 introduction. After a 3-day first-phase improvement (cessation of neuromuscular blocking agents and reduction of sedation), this subject subsequently developed several ventilator-associated pneumonias and ultimately died 1 month later.

[0218] 7.1.2.5. Discussion CER-001 not only showed very good tolerance in the acute phase, but also, in parallel with the normalization of ApoA-I levels after CER-001 administration, rapid improvement in the respiratory state, reduction of inflammatory parameters, and normalization of blood cell counts were observed in 3 out of 4 subjects. These subjects developed severe COVID-19-related cytokine storms, but were able to be discharged without oxygen assistance only 3 to 4 days after the infusion of CER-001. In 3 subjects who had a favorable outcome after CER-001 administration, a rapid decrease in IL-8 was observed in parallel with clinical and biological improvements. In Subject 4, after a first-phase clinical improvement accompanied by normalization of ApoA-I and decrease in IL-8, ventilator-associated pneumonia and clinical deterioration were accompanied by increases in C-reactive protein (CRP) and IL-8, as well as a decrease in ApoA-I. In this study, the subjects received 4 infusions of CER-001 (10 mg / kg), but it is thought that using a higher dose (e.g., 15 mg / kg) in the first injection would help ApoA-I and non-oxidized HDL reach optimal concentrations more rapidly and achieve the maximum therapeutic effect.

[0219] Although not bound by theory, the results of this study are believed to be extensible to other situations of hyperinflammatory states, such as virus-induced hyperinflammatory states (e.g., virus-induced HLH, dengue hemorrhagic fever, dengue shock syndrome, herpes simplex virus infection, etc.), and other forms of HLH (e.g., familial HLH, and HLH subsequent to other conditions such as acute leukemia and lymphoma).

[0220] 8. Specific Embodiments Various aspects of the present disclosure are described in the embodiments set forth in the following numbered paragraphs. 1. A method of treating a subject having or at risk of having a hyperinflammatory state, comprising administering to the subject a predetermined dose of a lipid-binding protein-based complex, optionally, wherein the hyperinflammatory state is hemophagocytic lymphohistiocytosis (HLH), dengue hemorrhagic fever, or dengue shock syndrome. 2. A method of treating a subject having or at risk of having hemophagocytic lymphohistiocytosis (HLH), comprising administering to the subject a predetermined dose of a lipid-binding protein-based complex. 3. The method according to embodiment 1 or 2, wherein the subject has or is at risk of having HLH subsequent to a non-malignant condition. 4. The method according to embodiment 3, wherein the non-malignant condition is a viral infection. 5. The method according to embodiment 4, wherein the subject has or is at risk of having HLH caused by a dengue fever infection. 6. The method according to embodiment 5, wherein the subject has dengue fever. 7. The method according to embodiment 5, wherein the subject has dengue hemorrhagic fever. 8. The method according to embodiment 5, wherein the subject has dengue shock syndrome. 9. The method according to embodiment 4, wherein the subject has or is at risk of having HLH caused by a herpes simplex virus infection. 10. The method according to embodiment 4, wherein the subject has or is at risk of having HLH caused by an Epstein-Barr virus infection. 11. The method according to embodiment 3, wherein the non-pathological condition is an autoimmune disease. 12. The method according to embodiment 1 or 2, wherein the subject has or is at risk of having HLH subsequent to the pathological condition. 13. The method according to embodiment 12, wherein the pathological condition is leukemia or lymphoma. 14. The method according to embodiment 1 or 2, wherein the subject has or is at risk of having familial HLH. 15. The method according to any one of embodiments 1 to 14, wherein the subject has HLH. 16. The method according to any one of embodiments 1 to 14, wherein the subject is at risk of HLH. 17. A method of treating a subject having a dengue infection, the method comprising administering to the subject a predetermined dose of a lipid-bound protein-based complex. 18. The method according to embodiment 1 or 17, wherein the subject has dengue. 19. The method according to embodiment 1 or 17, wherein the subject has dengue hemorrhagic fever. 20. The method according to embodiment 1 or 17, wherein the subject is at risk of dengue hemorrhagic fever. 21. The method according to embodiment 1 or 17, wherein the subject has dengue shock syndrome. 22. The method according to embodiment 1 or 17, wherein the subject is at risk of dengue shock syndrome. 23. A method of treating a subject having a herpes simplex infection, the method comprising administering to the subject a predetermined dose of a lipid-bound protein-based complex. 24. A method of treating a subject having an Epstein-Barr infection, the method comprising administering to the subject a predetermined dose of a lipid-bound protein-based complex. 25. The method according to any one of embodiments 1 to 24, wherein the dose is a high dose. 26. The method according to embodiment 25, wherein the high dose is administered over a period of 1 day to approximately 2 weeks, and optionally, the high dose is administered over a period of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days. 27. The method according to embodiment 25 or embodiment 26, wherein the high dose is a set of 2 to 10 individual doses, and optionally, the high dose is a set of 3, 4, 5, 6, 7, 8, 9, or 10 individual doses. 28. The method according to embodiment 27, wherein the plurality of individual doses are administered daily or twice a day. 29. The method according to embodiment 27 or embodiment 28, wherein the plurality of individual doses are administered at intervals of 2 to 3 days. 30. The method according to embodiment 27, wherein the plurality of individual doses are administered at intervals within 1 day. 31. The method according to embodiment 30, comprising administering two or more individual doses at approximately 12-hour intervals. 32. The method according to embodiment 31, comprising administering two individual doses at approximately 12-hour intervals. 33. The method according to embodiment 31, comprising administering three individual doses at approximately 12-hour intervals. 34. The method according to embodiment 32 or embodiment 33, further comprising administering an individual dose approximately 1 day later. 35. The method according to embodiment 27, comprising administering three individual doses at approximately 12-hour intervals and a fourth individual dose approximately 1 day later. 36. The method according to embodiment 25, wherein the high dose is administered as a single individual dose. 37. The method according to embodiment 25, wherein the high dose is a set of two individual doses administered within 1 day. 38. The method according to embodiment 37, wherein the two individual doses are administered at approximately 12-hour intervals. 39. The method according to any one of embodiments 27 to 38, wherein each individual dose is effective to increase the HDL level of the subject. 40. The method according to embodiment 39, wherein a high dose is effective to increase the serum HDL level of the subject to a normal value (e.g., > 0.45 g / L). 41. The method according to embodiment 39, wherein each individual dose is effective to increase the HDL level of the subject by at least 25%, at least 30%, or at least 35% 2 to 4 hours after administration. 42. The method according to embodiment 41, wherein each individual dose is effective to increase the HDL level of the subject by at least 25%, at least 30%, or at least 35% 2 hours after administration. 43. The method according to embodiment 41, wherein each individual dose is effective to increase the HDL level of the subject by at least 25%, at least 30%, or at least 35% 3 hours after administration. 44. The method according to embodiment 41, wherein each individual dose is effective to increase the HDL level of the subject by at least 25%, at least 30%, or at least 35% 4 hours after administration. 45. The method according to any one of embodiments 27 to 44, wherein each individual dose is effective to increase the ApoA-I level of the subject. 46. The method according to embodiment 45, wherein a high dose is effective to increase the serum ApoA-I level of the subject to a normal value (e.g., > 1.1 g / L). 47. The method according to embodiment 45, wherein each individual dose is effective to increase the ApoA-I level of the subject by at least 25%, at least 30%, or at least 35% 2 to 4 hours after administration. 48. The method according to embodiment 46, wherein each individual dose is effective to increase the ApoA-I level of the subject by at least 25%, at least 30%, or at least 35% 2 hours after administration. 49. The method according to embodiment 46, wherein each individual dose is effective to increase the ApoA-I level of the subject by at least 25%, at least 30%, or at least 35% 3 hours after administration. 50. The method according to embodiment 46, wherein each individual dose is effective to increase the subject's ApoA-I level by at least 25%, at least 30%, or at least 35% 4 hours after administration. 51. The method according to any one of embodiments 25 to 50, wherein the high dose is effective to improve the subject's vascular endothelial function, and optionally, the vascular endothelial function is measured by circulating VCAM-1 and / or ICAM-1. 52. The method according to any one of embodiments 25 to 51, wherein the high dose is effective to reduce the serum level of one or more inflammatory markers in the subject. 53. The method according to embodiment 52, wherein the high dose is effective to reduce the serum level of interleukin-6 ("IL-6"). 54. The method according to embodiment 52 or embodiment 53, wherein the high dose is effective to reduce the serum level of C-reactive protein. 55. The method according to any one of embodiments 52 to 54, wherein the high dose is effective to reduce the serum level of D-dimer. 56. The method according to any one of embodiments 52 to 55, wherein the high dose is effective to reduce the serum level of ferritin. 57. The method according to any one of embodiments 52 to 56, wherein the high dose is effective to reduce the serum level of interleukin 8 (IL-8). 58. The method according to any one of embodiments 52 to 56, wherein the high dose is effective to normalize the serum level of interleukin 8 (IL-8). 59. The method according to any one of embodiments 52 to 58, wherein the high dose is effective to reduce the serum level of granulocyte macrophage colony-stimulating factor (GM-CSF). 60. The method according to any one of embodiments 52 to 59, wherein the high dose is effective to reduce the serum level of monocyte chemoattractant protein (MCP) 1. 61. The method according to any one of embodiments 52 to 60, wherein the high dose is effective for reducing the serum level of tumor necrosis factor α (TNF-α). 62. The method according to any one of embodiments 52 to 61, wherein the high dose is effective for reducing the serum level of one or more inflammatory markers from an elevated range to a normal range. 63. The method according to any one of embodiments 52 to 62, wherein the high dose is effective for reducing the serum level of one or more inflammatory markers by at least 20%, at least 40%, or at least 60%. 64. The method according to any one of embodiments 1 to 63, wherein the subject has or is at risk of CRS. 65. The method according to embodiment 64, wherein the subject has CRS. 66. The method according to embodiment 64, wherein the subject is at risk of CRS. 67. The method according to any one of embodiments 25 to 66, wherein the high dose is effective for reducing the likelihood that the subject will develop acute kidney injury (AKI). 68. The method according to any one of embodiments 25 to 67, wherein the high dose is effective for delaying the onset of AKI. 69. The method according to any one of embodiments 25 to 67, wherein the high dose is effective for preventing AKI. 70. The method according to any one of embodiments 25 to 66, wherein the subject has or is at risk of developing acute kidney injury (AKI). 71. The method according to embodiment 70, wherein the subject has AKI. 72. The method according to embodiment 71, wherein the high dose is effective for reducing the severity of AKI. 73. The method according to embodiment 70, wherein the subject is at risk of AKI. 74. The method according to embodiment 73, wherein the high dose is effective for reducing the likelihood that the subject will develop AKI. 75. The method according to embodiment 73, wherein the high dose is effective for delaying the onset of AKI. 76. The method according to embodiment 73, wherein a high dose is effective in preventing AKI. 77. The method according to embodiment 73, wherein a high dose is effective in reducing the severity of AKI when the subject develops AKI. 78. The method according to any one of embodiments 1 to 77, wherein the subject has a SOFA score of 1 to 4 before administration of the complex based on the lipid-binding protein. 79. The method according to embodiment 78, wherein the subject has a SOFA score of 2 to 4 before administration of the complex based on the lipid-binding protein. 80. The method according to embodiment 78, wherein the subject has a SOFA score of 1 before administration of the complex based on the lipid-binding protein. 81. The method according to embodiment 78, wherein the subject has a SOFA score of 2 before administration of the complex based on the lipid-binding protein. 82. The method according to embodiment 78, wherein the subject has a SOFA score of 3 before administration of the complex based on the lipid-binding protein. 83. The method according to embodiment 78, wherein the subject has a SOFA score of 4 before administration of the complex based on the lipid-binding protein. 84. The method according to any one of embodiments 1 to 83, wherein the complex based on the lipid-binding protein is reconstituted HDL or an HDL mimetic. 85. The method according to any one of embodiments 1 to 83, wherein the complex based on the lipid-binding protein is an Apomer or a Cargomer. 86. The method according to any one of embodiments 1 to 85, wherein the complex based on the lipid-binding protein contains sphingomyelin. 87. The method according to any one of embodiments 1 to 86, wherein the complex based on the lipid-binding protein contains a negatively charged lipid. 88. The method according to embodiment 87, wherein the negatively charged lipid is 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (DPPG) or a salt thereof. 89. The method according to embodiment 84, wherein the complex based on the lipid-binding protein is CER-001, CSL-111, CSL-112, CER-522, or ETC-216. 90. The method according to embodiment 89, wherein the complex based on the lipid-binding protein is CER-001. 91. The method according to any one of embodiments 1 to 90, wherein the complex based on the lipid-binding protein is administered systemically, optionally by infusion. 92. The method according to any one of embodiments 1 to 91, wherein the complex based on the lipid-binding protein is administered until the serum level of one or more inflammatory markers decreases. 93. The method according to embodiment 92, wherein the complex based on the lipid-binding protein is administered until the serum level of one or more inflammatory markers decreases to the normal range(s). 94. The method according to embodiment 92, wherein the complex based on the lipid-binding protein is administered until the serum level of one or more inflammatory markers decreases below the baseline level(s) for the one or more inflammatory markers measured prior to administration of the complex based on the lipid-binding protein. 95. The method according to any one of embodiments 1 to 94, wherein each individual dose of the complex based on the lipid-binding protein is 4 - 40 mg / kg (protein weight basis). 96. The method according to embodiment 95, wherein each individual dose of the complex based on the lipid-binding protein is 4 - 30 mg / kg (protein weight basis). 97. The method according to embodiment 95, wherein each individual dose of the complex based on the lipid-binding protein is 15 - 25 mg / kg (protein weight basis). 98. The method according to embodiment 95, wherein each individual dose of the complex based on the lipid-binding protein is 10 - 30 mg / kg (protein weight basis). 99. The method according to embodiment 95, wherein each individual dose of the complex based on the lipid-binding protein is 10 - 20 mg / kg (protein weight basis). 100. The method according to embodiment 95, wherein the individual dose of each complex based on the lipid-binding protein is 5 mg / kg (protein weight basis). 101. The method according to embodiment 95, wherein the individual dose of each complex based on the lipid-binding protein is 10 mg / kg (protein weight basis). 102. The method according to embodiment 95, wherein the individual dose of each complex based on the lipid-binding protein is 15 mg / kg (protein weight basis). 103. The method according to embodiment 95, wherein the individual dose of each complex based on the lipid-binding protein is 20 mg / kg (protein weight basis). 104. The method according to embodiment 95, wherein the individual dose of each complex based on the lipid-binding protein is 5 - 15 mg / kg (protein weight basis). 105. The method according to embodiment 95, wherein the individual dose of each complex based on the lipid-binding protein is 10 - 20 mg / kg (protein weight basis). 106. The method according to embodiment 95, wherein the individual dose of each complex based on the lipid-binding protein is 15 - 25 mg / kg (protein weight basis). 107. The method according to any one of embodiments 25 to 106, wherein the high dose is administered according to an induction regimen, optionally followed by a consolidation therapy regimen. 108. The method according to embodiment 107, wherein the induction regimen comprises administering the complex based on the lipid-binding protein once or twice a day. 109. The method according to embodiment 107 or 108, wherein the consolidation therapy regimen comprises administering the complex based on the lipid-binding protein once a day or once every two days. 110. The method according to any one of embodiments 1 to 109, wherein the subject is not treated with a maintenance regimen. 111. The method according to any one of embodiments 107 to 110, wherein the consolidation therapy regimen comprises administering to the subject, one or more doses of a lipid-binding protein-based complex, one day or several days after administration of the final dose of the induction regimen. 112. The method according to embodiment 111, wherein the first dose of the lipid-binding protein-based complex administered in the consolidation therapy regimen is administered more than 2 days after administration of the final dose of the induction regimen. 113. The method according to embodiment 111, wherein the first dose of the lipid-binding protein-based complex administered in the consolidation therapy regimen is administered more than 3 days after administration of the final dose of the induction regimen. 114. The method according to embodiment 113, wherein the first dose of the lipid-binding protein-based complex administered in the consolidation therapy regimen is administered 3 days after administration of the final dose of the induction regimen. 115. The method according to any one of embodiments 107 to 114, comprising an induction regimen including administration of the lipid-binding protein-based complex twice a day on days 1, 2, and 3, and a consolidation therapy regimen including two doses of the lipid-binding protein-based complex on day 6. 116. The method according to any one of embodiments 107 to 115, wherein each individual dose of the lipid-binding protein-based complex administered in the induction regimen is 4 - 40 mg / kg (protein weight basis). 117. The method according to any one of embodiments 107 to 116, wherein each individual dose of the lipid-binding protein-based complex administered in the induction regimen is 4 - 30 mg / kg (protein weight basis). 118. The method according to any one of embodiments 107 to 116, wherein each individual dose of the lipid-binding protein-based complex administered in the induction regimen is 15 - 25 mg / kg (protein weight basis). 119. The method according to any one of embodiments 107 to 116, wherein each individual dose of the lipid-binding protein-based complex administered in the induction regimen is 10 - 30 mg / kg (protein weight basis). 120. The method according to any one of embodiments 107 to 116, wherein the individual dose of each lipid-binding protein-based complex administered in the induction regimen is 10 to 20 mg / kg (protein weight basis). 121. The method according to any one of embodiments 107 to 116, wherein the individual dose of each lipid-binding protein-based complex administered in the induction regimen is 5 mg / kg (protein weight basis). 122. The method according to any one of embodiments 107 to 116, wherein the individual dose of each lipid-binding protein-based complex administered in the induction regimen is 10 mg / kg (protein weight basis). 123. The method according to any one of embodiments 107 to 116, wherein the individual dose of each lipid-binding protein-based complex administered in the induction regimen is 15 mg / kg (protein weight basis). 124. The method according to any one of embodiments 107 to 116, wherein the individual dose of each lipid-binding protein-based complex administered in the induction regimen is 20 mg / kg (protein weight basis). 125. The method according to any one of embodiments 107 to 124, wherein the dose of the lipid-binding protein-based complex administered in the consolidation therapy regimen is 5 to 15 mg / kg (protein weight basis). 126. The method according to any one of embodiments 107 to 124, wherein the dose of the lipid-binding protein-based complex administered in the consolidation therapy regimen is 10 to 20 mg / kg (protein weight basis). 127. The method according to any one of embodiments 107 to 124, wherein the dose of the lipid-binding protein-based complex administered in the consolidation therapy regimen is 15 to 25 mg / kg (protein weight basis). 128. The method according to any one of embodiments 107 to 124, wherein the dose of the lipid-binding protein-based complex administered in the consolidation therapy regimen is 5 mg / kg (protein weight basis). 129. The method according to any one of embodiments 107 to 124, wherein the dosage of the complex based on the lipid-binding protein administered in the soil consolidation therapy regimen is 10 mg / kg (protein weight basis). 130. The method according to any one of embodiments 107 to 124, wherein the dosage of the complex based on the lipid-binding protein administered in the soil consolidation therapy regimen is 15 mg / kg (protein weight basis). 131. The method according to any one of embodiments 1 to 130, wherein each individual dosage of the complex based on the lipid-binding protein to be administered is 300 mg to 4000 mg (protein weight basis). 132. The method according to embodiment 131, wherein each individual dosage of the complex based on the lipid-binding protein to be administered is 300 mg to 3000 mg (protein weight basis). 133. The method according to embodiment 131, wherein each individual dosage of the complex based on the lipid-binding protein to be administered is 300 mg to 1500 mg (protein weight basis). 134. The method according to embodiment 131, wherein each individual dosage of the complex based on the lipid-binding protein to be administered is 400 mg to 4000 mg (protein weight basis). 135. The method according to embodiment 131, wherein each individual dosage of the complex based on the lipid-binding protein to be administered is 400 mg to 1500 mg (protein weight basis). 136. The method according to embodiment 131, wherein each individual dosage of the complex based on the lipid-binding protein to be administered is 500 mg to 1200 mg (protein weight basis). 137. The method according to embodiment 131, wherein each individual dosage of the complex based on the lipid-binding protein to be administered is 500 mg to 1000 mg (protein weight basis). 138. The method according to embodiment 131, wherein each individual dosage of the complex based on the lipid-binding protein to be administered is 600 mg to 3000 mg (protein weight basis). 139. The method according to embodiment 131, wherein the individual dose of each complex based on the lipid-binding protein to be administered is 800 mg to 3000 mg (protein weight basis). 140. The method according to embodiment 131, wherein the individual dose of each complex based on the lipid-binding protein to be administered is 1000 mg to 2400 mg (protein weight basis). 141. The method according to embodiment 131, wherein the individual dose of each complex based on the lipid-binding protein to be administered is 1000 mg to 2000 mg (protein weight basis). 142. The method according to any one of embodiments 25 to 141, wherein the high dose of the complex based on the lipid-binding protein is 600 mg to 40 g (protein weight basis). 143. The method according to any one of embodiments 25 to 141, wherein the high dose of the complex based on the lipid-binding protein is 3 g to 35 g (protein weight basis). 144. The method according to any one of embodiments 25 to 141, wherein the high dose of the complex based on the lipid-binding protein is 5 g to 30 g (protein weight basis). 145. The method according to any one of embodiments 1 to 144, wherein the complex based on the lipid-binding protein is administered by infusion. 146. The method according to embodiment 145, wherein each individual dose is administered over a period of 1 to 24 hours. 147. The method according to embodiment 146, wherein each individual dose is administered over a period of 24 hours. 148. The method according to embodiment 145, wherein each individual dose is administered over a period of 1 hour or less. 149. The method according to embodiment 145, wherein each individual dose is administered over a period of 0.5 hour to 1 hour. 150. The method according to any one of embodiments 1 to 149, further comprising administering an antihistamine to the subject prior to each individual dose. 151. The method according to embodiment 150, wherein the antihistamine comprises dechlorpheniramine or hydroxyzine. 152. The method according to any one of embodiments 1 to 151, wherein the subject has received or is receiving one or more additional treatments and / or further comprises administering to the subject one or more additional treatments. 153. The method according to embodiment 152, wherein the one or more additional treatments comprise one or more anti-IL-6 agents. 154. The method according to embodiment 153, wherein the one or more anti-IL-6 agents comprise tocilizumab, siltuximab, olaratumab, elsilimomab, BMS-945429, sirukumab, revilimab, CPSI-2364, or a combination thereof. 155. The method according to embodiment 154, wherein the one or more anti-IL-6 agents comprise tocilizumab. 156. The method according to any one of embodiments 152 to 155, wherein the one or more additional treatments comprise one or more corticosteroids. 157. The method according to embodiment 156, wherein the one or more corticosteroids comprise methylprednisolone, dexamethasone, or a combination thereof. 158. The method according to any one of embodiments 1 to 157, wherein the lipid-binding protein-based complex is CER-001. 159. The method according to embodiment 158, wherein CER-001 is a lipoprotein complex comprising ApoA-I and phospholipids at a ratio of ApoA-I weight:total phospholipid weight of 1:2.7 ± 20% and sphingomyelin phospholipid and DPPG at a ratio of sphingomyelin weight:DPPG weight of 97:3 ± 20%. 160. The method according to embodiment 158, wherein CER-001 is a lipoprotein complex comprising ApoA-I and phospholipids at a ratio of ApoA-I weight:total phospholipid weight of 1:2.7 ± 10% and sphingomyelin phospholipid and DPPG at a ratio of sphingomyelin weight:DPPG weight of 97:3 ± 10%. 161. The method according to embodiment 158, wherein CER-001 is a lipoprotein complex comprising ApoA-I and phospholipids at a ratio of ApoA-I weight:total phospholipid weight of 1:2.7, and sphingomyelin and DPPG at a weight:weight ratio of 97:3 of phospholipid sphingomyelin and DPPG. 162. The method according to any one of embodiments 159 to 161, wherein ApoA-I has the amino acid sequence of amino acids 25 to 267 of SEQ ID NO: 2. 163. The method according to any one of embodiments 159 to 162, wherein ApoA-I is recombinantly expressed. 164. The method according to any one of embodiments 159 to 163, wherein CER-001 contains natural sphingomyelin. 165. The method according to embodiment 164, wherein the natural sphingomyelin is chicken egg sphingomyelin. 166. The method according to any one of embodiments 159 to 163, wherein CER-001 contains synthetic sphingomyelin. 167. The method according to embodiment 166, wherein the synthetic sphingomyelin is palmitoyl sphingomyelin. 168. The method according to any one of embodiments 158 to 167, wherein CER-001 is administered in the form of a formulation in which CER-001 is at least 95% homogeneous. 169. The method according to embodiment 168, wherein CER-001 is administered in the form of a formulation in which CER-001 is at least 97% homogeneous. 170. The method according to embodiment 168, wherein CER-001 is administered in the form of a formulation in which CER-001 is at least 98% homogeneous. 171. The method according to embodiment 168, wherein CER-001 is administered in the form of a formulation in which CER-001 is at least 99% homogeneous. 172. The method according to any one of embodiments 1 to 171, wherein the subject is human. 173. The method according to any one of embodiments 1 to 172, wherein when CER-001 is first administered, the subject is not receiving mechanical ventilation. 174. The method according to embodiment 89, wherein the complex based on the lipid-binding protein is CSL-112. 175. The method according to any one of embodiments 163 to 173, wherein ApoA-I is produced by a mammalian host cell. 176. The method according to embodiment 175, wherein the mammalian host cell is a Chinese hamster ovary (CHO) cell. 177. The method according to embodiment 176, wherein the CHO cell is a CHO-S cell. 178. The method according to any one of embodiments 175 to 177, wherein ApoA-I has undergone post-translational processing (e.g., glycosylation) such that ApoA-I has one or more structural features (e.g., glycosylation pattern) different from human ApoA-I purified from human plasma. 179. A method of treating a subject having or at risk of having a hyperinflammatory condition, comprising administering to the subject a predetermined dose of an apolipoprotein A-I ("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, and optionally, the hyperinflammatory condition is hemophagocytic lymphohistiocytosis (HLH), dengue hemorrhagic fever, or dengue shock syndrome. 180. A method of treating a subject having or at risk of having hemophagocytic lymphohistiocytosis (HLH), comprising administering to the subject a predetermined dose of an apolipoprotein A-I ("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. 181. The method according to embodiment 179 or embodiment 180, wherein the subject has or is at risk of having HLH subsequent to a non-pathological condition. 182. The method according to embodiment 181, wherein the non-pathological condition is a viral infection. 183. The method according to embodiment 182, wherein the subject has or is at risk of having HLH resulting from a dengue infection. 184. The method according to embodiment 183, wherein the subject has dengue fever. 185. The method according to embodiment 183, wherein the subject has dengue hemorrhagic fever. 186. The method according to embodiment 183, wherein the subject has dengue shock syndrome. 187. The method according to embodiment 182, wherein the subject has or is at risk of having HLH caused by herpes simplex infection. 188. The method according to embodiment 182, wherein the subject has or is at risk of having HLH caused by Epstein - Barr virus infection. 189. The method according to embodiment 181, wherein the non - adverse condition is an autoimmune disease. 190. The method according to embodiment 179 or embodiment 180, wherein the subject has or is at risk of having HLH subsequent to an adverse condition. 191. The method according to embodiment 190, wherein the adverse condition is leukemia or lymphoma. 192. The method according to embodiment 179 or embodiment 180, wherein the subject has or is at risk of having familial HLH. 193. The method according to any one of embodiments 179 to 192, wherein the subject has HLH. 194. The method according to any one of embodiments 179 to 192, wherein the subject is at risk of HLH. 195. A method of treating a subject having a dengue fever infection, comprising administering to the subject a predetermined dose of an apolipoprotein A - I ( "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. 196. The method according to embodiment 179 or embodiment 195, wherein the subject has dengue fever. 197. The method according to embodiment 179 or embodiment 195, wherein the subject has dengue hemorrhagic fever. 198. The method according to embodiment 179 or embodiment 195, wherein the subject is at risk of dengue hemorrhagic fever. 199. The method according to embodiment 179 or embodiment 195, wherein the subject has dengue shock syndrome. 200. The method according to embodiment 179 or embodiment 195, wherein the subject has a risk of dengue shock syndrome. 201. A method for treating a subject having a herpes simplex infection, comprising administering to the subject a predetermined dose of an apolipoprotein A-I (''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. 202. A method for treating a subject having an Epstein-Barr infection, comprising administering to the subject a predetermined dose of an apolipoprotein A-I (''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. 203. The method according to any one of embodiments 179 to 202, wherein the dose is a high dose. 204. The method according to embodiment 203, wherein the high dose is administered over a period of 1 day to approximately 2 weeks, and optionally, the high dose is administered over a period of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days. 205. The method according to embodiment 203 or embodiment 204, wherein the high dose is a set of 2 to 10 individual doses, and optionally, the high dose is a set of 3, 4, 5, 6, 7, 8, 9, or 10 individual doses. 206. The method according to embodiment 205, wherein the plurality of individual doses are administered daily or twice a day. 207. The method according to embodiment 205 or embodiment 206, wherein the plurality of individual doses are administered at intervals of 2 to 3 days. 208. The method according to embodiment 205, wherein the plurality of individual doses are administered at intervals within 1 day. 209. The method according to embodiment 208, comprising administering two or more individual doses at intervals of approximately 12 hours. 210. The method according to embodiment 209, comprising administering two individual doses at intervals of approximately 12 hours. The method according to embodiment 209, comprising administering three separate doses at approximately 12-hour intervals. The method according to embodiment 210 or embodiment 211, further comprising administering the separate doses approximately one day apart. The method according to embodiment 205, comprising administering three separate doses at approximately 12-hour intervals and a fourth separate dose approximately one day later. The method according to embodiment 203, wherein the high dose is administered as a single separate dose. The method according to embodiment 203, wherein the high dose is a set of two separate doses administered within one day. The method according to embodiment 215, wherein the two separate doses are administered at approximately 12-hour intervals. The method according to any one of embodiments 205 to 216, wherein each separate dose is effective to increase the HDL level of the subject. The method according to embodiment 217, wherein the high dose is effective to increase the serum HDL level of the subject to a normal value (e.g., >0.45 g / L). The method according to embodiment 217, wherein each separate dose is effective to increase the HDL level of the subject by at least 25%, at least 30%, or at least 35% two to four hours after administration. The method according to embodiment 219, wherein each separate dose is effective to increase the HDL level of the subject by at least 25%, at least 30%, or at least 35% two hours after administration. The method according to embodiment 219, wherein each separate dose is effective to increase the HDL level of the subject by at least 25%, at least 30%, or at least 35% three hours after administration. The method according to embodiment 219, wherein each separate dose is effective to increase the HDL level of the subject by at least 25%, at least 30%, or at least 35% four hours after administration. 223. The method according to any one of embodiments 205 to 222, wherein each individual dose is effective to increase the ApoA-I level of the subject. 224. The method according to embodiment 223, wherein the high dose is effective to increase the serum ApoA-I level of the subject to a normal value (e.g., >1.1 g / L). 225. The method according to embodiment 223, wherein each individual dose is effective to increase the ApoA-I level of the subject by at least 25%, at least 30%, or at least 35% at 2 to 4 hours after administration. 226. The method according to embodiment 224, wherein each individual dose is effective to increase the ApoA-I level of the subject by at least 25%, at least 30%, or at least 35% at 2 hours after administration. 227. The method according to embodiment 224, wherein each individual dose is effective to increase the ApoA-I level of the subject by at least 25%, at least 30%, or at least 35% at 3 hours after administration. 228. The method according to embodiment 224, wherein each individual dose is effective to increase the ApoA-I level of the subject by at least 25%, at least 30%, or at least 35% at 4 hours after administration. 229. The method according to any one of embodiments 203 to 228, wherein the high dose is effective to improve the endothelial function of the blood vessels of the subject, and optionally, the endothelial function of the blood vessels is measured by circulating VCAM-1 and / or ICAM-1. 230. The method according to any one of embodiments 203 to 229, wherein the high dose is effective to reduce the serum level of one or more inflammatory markers in the subject. 231. The method according to embodiment 230, wherein the high dose is effective to reduce the serum level of interleukin-6 ("IL-6"). 232. The method according to embodiment 230 or embodiment 231, wherein the high dose is effective to reduce the serum level of C-reactive protein. 233. The method according to any one of embodiments 230 to 232, wherein the high dose is effective in reducing the serum level of D-dimer. 234. The method according to any one of embodiments 230 to 233, wherein the high dose is effective in reducing the serum level of ferritin. 235. The method according to any one of embodiments 230 to 234, wherein the high dose is effective in reducing the serum level of interleukin 8 (IL-8). 236. The method according to any one of embodiments 230 to 234, wherein the high dose is effective in normalizing the serum level of interleukin 8 (IL-8). 237. The method according to any one of embodiments 230 to 236, wherein the high dose is effective in reducing the serum level of granulocyte macrophage colony-stimulating factor (GM-CSF). 238. The method according to any one of embodiments 230 to 237, wherein the high dose is effective in reducing the serum level of monocyte chemoattractant protein (MCP) 1. 239. The method according to any one of embodiments 230 to 238, wherein the high dose is effective in reducing the serum level of tumor necrosis factor α (TNF-α). 240. The method according to any one of embodiments 230 to 239, wherein the high dose is effective in reducing the serum level of one or more inflammatory markers from an elevated range to a normal range. 241. The method according to any one of embodiments 230 to 240, wherein the high dose is effective in reducing the serum level of one or more inflammatory markers by at least 20%, at least 40%, or at least 60%. 242. The method according to any one of embodiments 179 to 241, wherein the subject has CRS or is at risk of CRS. 243. The method according to embodiment 242, wherein the subject has CRS. 244. The method according to embodiment 242, wherein the subject is at risk of CRS. 245. The method according to any one of embodiments 203 to 244, wherein the high dose is effective in reducing the likelihood that the subject will develop acute kidney injury (AKI). 246. The method according to any one of embodiments 203 to 245, wherein the high dose is effective in delaying the onset of AKI. 247. The method according to any one of embodiments 203 to 245, wherein the high dose is effective in preventing AKI. 248. The method according to any one of embodiments 203 to 244, wherein the subject has or is at risk of developing acute kidney injury (AKI). 249. The method according to embodiment 248, wherein the subject has AKI. 250. The method according to embodiment 249, wherein the high dose is effective in reducing the severity of AKI. 251. The method according to embodiment 248, wherein the subject is at risk of AKI. 252. The method according to embodiment 251, wherein the high dose is effective in reducing the likelihood that the subject will develop AKI. 253. The method according to embodiment 251, wherein the high dose is effective in delaying the onset of AKI. 254. The method according to embodiment 251, wherein the high dose is effective in preventing AKI. 255. The method according to embodiment 251, wherein when the subject develops AKI, the high dose is effective in reducing the severity of AKI. 256. The method according to any one of embodiments 179 to 255, wherein the subject has a SOFA score of 1 to 4 prior to administration of the formulation. 257. The method according to embodiment 256, wherein the subject has a SOFA score of 2 to 4 prior to administration of the formulation. 258. The method according to embodiment 256, wherein the subject has a SOFA score of 1 prior to administration of the formulation. 259. The method according to embodiment 256, wherein the subject has a SOFA score of 2 prior to administration of the formulation. 260. The method according to embodiment 256, wherein the subject has a SOFA score of 3 prior to administration of the formulation. 261. The method according to embodiment 256, wherein the subject has a SOFA score of 4 before administration of the formulation. 262. The method according to any one of embodiments 179 to 261, wherein the formulation is reconstituted HDL or an HDL mimetic. 263. The method according to any one of embodiments 179 to 261, wherein the formulation is an Apomer or a Cargomer. 264. The method according to any one of embodiments 179 to 263, wherein the formulation contains sphingomyelin. 265. The method according to any one of embodiments 179 to 264, wherein the formulation contains a charged lipid. 266. The method according to embodiment 265, wherein the charged lipid is 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (DPPG) or a salt thereof. 267. The method according to any one of embodiments 179 to 266, wherein the formulation is optionally administered systemically by infusion. 268. The method according to any one of embodiments 179 to 267, wherein the formulation is administered until the serum level of one or more inflammatory markers decreases. 269. The method according to embodiment 268, wherein the formulation is administered until the serum level of one or more inflammatory markers decreases to the normal range(s). 270. The method according to embodiment 268, wherein the formulation is administered until the serum level of one or more inflammatory markers decreases below the baseline level(s) for one or more inflammatory markers measured prior to administration of the lipid-bound protein-based complex. 271. The method according to any one of embodiments 179 to 270, wherein each individual dose of the formulation administered is 4 - 40 mg / kg (protein weight basis). 272. The method according to embodiment 271, wherein each individual dose of the formulation is 4 - 30 mg / kg (protein weight basis). 273. The method according to embodiment 271, wherein each individual dose of the formulation is 15 - 25 mg / kg (protein weight basis). 274. The method according to embodiment 271, wherein the individual dose of each formulation is 10 to 30 mg / kg (protein weight basis). 275. The method according to embodiment 271, wherein the individual dose of each formulation is 10 to 20 mg / kg (protein weight basis). 276. The method according to embodiment 271, wherein the individual dose of each formulation is 5 mg / kg (protein weight basis). 277. The method according to embodiment 271, wherein the individual dose of each formulation is 10 mg / kg (protein weight basis). 278. The method according to embodiment 271, wherein the individual dose of each formulation is 15 mg / kg (protein weight basis). 279. The method according to embodiment 271, wherein the individual dose of each formulation is 20 mg / kg (protein weight basis). 280. The method according to embodiment 271, wherein the individual dose of each formulation is 5 to 15 mg / kg (protein weight basis). 281. The method according to embodiment 271, wherein the individual dose of each formulation is 10 to 20 mg / kg (protein weight basis). 282. The method according to embodiment 271, wherein the individual dose of each formulation is 15 to 25 mg / kg (protein weight basis). 283. The method according to any one of embodiments 203 to 282, wherein the high dose is administered according to an induction regimen, and optionally, a consolidation therapy regimen follows it. 284. The method according to embodiment 283, wherein the induction regimen comprises administering the formulation once a day or twice a day. 285. The method according to embodiment 283 or embodiment 284, wherein the consolidation therapy regimen comprises administering the formulation once a day or once every two days. 286. The method according to any one of embodiments 179 to 285, wherein the subject is not treated with a maintenance regimen. The method according to any one of embodiments 283 to 286, wherein the consolidation therapy regimen comprises administering one or more doses of the formulation to the subject one day or several days after administering the final dose of the induction regimen. The method according to embodiment 287, wherein the first dose of the formulation administered during the consolidation therapy regimen is administered more than 2 days after administering the final dose of the induction regimen. The method according to embodiment 287, wherein the first dose of the formulation administered during the consolidation therapy regimen is administered more than 3 days after administering the final dose of the induction regimen. The method according to embodiment 287, wherein the first administration of the formulation administered during the consolidation therapy regimen is administered 3 days after administering the final dose of the induction regimen. The method according to any one of embodiments 283 to 290, comprising an induction regimen that includes administering the formulation twice a day on days 1, 2, and 3, and a consolidation therapy regimen that includes two doses of the formulation on day 6. The method according to any one of embodiments 283 to 291, wherein each individual dose of the formulation administered in the induction regimen is 4 - 40 mg / kg (protein weight basis). The method according to any one of embodiments 283 to 292, wherein each individual dose of the formulation administered in the induction regimen is 4 - 30 mg / kg (protein weight basis). The method according to any one of embodiments 283 to 292, wherein each individual dose of the formulation administered in the induction regimen is 15 - 25 mg / kg (protein weight basis). The method according to any one of embodiments 283 to 292, wherein each individual dose of the formulation administered in the induction regimen is 10 - 30 mg / kg (protein weight basis). The method according to any one of embodiments 283 to 292, wherein each individual dose of the formulation administered in the induction regimen is 10 - 20 mg / kg (protein weight basis). The method according to any one of embodiments 283 to 292, wherein each individual dose of the formulation administered in the induction regimen is 5 mg / kg (protein weight basis). The method according to any one of embodiments 283 to 292, wherein each individual dose of the formulation administered in the induction regimen is 10 mg / kg (protein weight basis). The method according to any one of embodiments 283 to 292, wherein each individual dose of the formulation administered in the induction regimen is 15 mg / kg (protein weight basis). The method according to any one of embodiments 283 to 292, wherein each individual dose of the formulation administered in the induction regimen is 20 mg / kg (protein weight basis). The method according to any one of embodiments 283 to 300, wherein the dose of the formulation administered in the consolidation therapy regimen is 5 - 15 mg / kg (protein weight basis). The method according to any one of embodiments 283 to 300, wherein the dose of the formulation administered in the consolidation therapy regimen is 10 - 20 mg / kg (protein weight basis). The method according to any one of embodiments 283 to 300, wherein the dose of the formulation administered in the consolidation therapy regimen is 15 - 25 mg / kg (protein weight basis). The method according to any one of embodiments 283 to 300, wherein the dose of the formulation administered in the consolidation therapy regimen is 5 mg / kg (protein weight basis). The method according to any one of embodiments 283 to 300, wherein the dose of the formulation administered in the consolidation therapy regimen is 10 mg / kg (protein weight basis). The method according to any one of embodiments 283 to 300, wherein the dose of the formulation administered in the consolidation therapy regimen is 15 mg / kg (protein weight basis). 307. The method according to any one of embodiments 179 to 306, wherein the individual dose of each preparation to be administered is 300 mg to 4000 mg (protein weight basis). 308. The method according to embodiment 307, wherein the individual dose of each preparation to be administered is 300 mg to 3000 mg (protein weight basis). 309. The method according to embodiment 307, wherein the individual dose of each preparation to be administered is 300 mg to 1500 mg (protein weight basis). 310. The method according to embodiment 307, wherein the individual dose of each preparation to be administered is 400 mg to 4000 mg (protein weight basis). 311. The method according to embodiment 307, wherein the individual dose of each preparation to be administered is 400 mg to 1500 mg (protein weight basis). 312. The method according to embodiment 307, wherein the individual dose of each preparation to be administered is 500 mg to 1200 mg (protein weight basis). 313. The method according to embodiment 307, wherein the individual dose of each preparation to be administered is 500 mg to 1000 mg (protein weight basis). 314. The method according to embodiment 307, wherein the individual dose of each preparation to be administered is 600 mg to 3000 mg (protein weight basis). 315. The method according to embodiment 307, wherein the individual dose of each preparation to be administered is 800 mg to 3000 mg (protein weight basis). 316. The method according to embodiment 307, wherein the individual dose of each preparation to be administered is 1000 mg to 2400 mg (protein weight basis). 317. The method according to embodiment 307, wherein the individual dose of each preparation to be administered is 1000 mg to 2000 mg (protein weight basis). 318. The method according to any one of embodiments 203 to 317, wherein the high dose of the preparation is 600 mg to 40 g (protein weight basis). 319. The method according to any one of embodiments 203 to 317, wherein the high dose of the formulation is 3 g to 35 g (protein weight basis). 320. The method according to any one of embodiments 203 to 317, wherein the high dose of the formulation is 5 g to 30 g (protein weight basis). 321. The method according to any one of embodiments 179 to 320, wherein the formulation is administered by infusion. 322. The method according to embodiment 321, wherein each individual dose is administered over a period of 1 to 24 hours. 323. The method according to embodiment 322, wherein each individual dose is administered over a period of 24 hours. 324. The method according to embodiment 321, wherein each individual dose is administered over a period of 1 hour or less. 325. The method according to embodiment 321, wherein each individual dose is administered over a period of 0.5 hour to 1 hour. 326. The method according to any one of embodiments 179 to 325, further comprising administering an antihistamine to the subject prior to each individual dose. 327. The method according to embodiment 326, wherein the antihistamine comprises dechlorpheniramine or hydroxyzine. 328. The method according to any one of embodiments 179 to 327, wherein the subject has received or is receiving one or more additional treatments, and further comprising administering one or more additional treatments to the subject. 329. The method according to embodiment 328, wherein the one or more additional treatments comprise one or more anti-IL-6 agents. 330. The method according to embodiment 329, wherein the one or more anti-IL-6 agents comprise tocilizumab, siltuximab, ocrelizumab, elsilimomab, BMS-945429, sirukumab, rilonacept, CPSI-2364, or a combination thereof. 331. The method according to embodiment 330, wherein the one or more anti-IL-6 agents comprise tocilizumab. 332. The method according to any one of embodiments 328 to 331, wherein one or more additional treatments comprise one or more corticosteroids. 333. The method according to embodiment 332, wherein the one or more corticosteroids comprise methylprednisolone, dexamethasone, or a combination thereof. 334. The method according to any one of embodiments 179 to 333, wherein the formulation comprises ApoA-I and phospholipids at an ApoA-I weight:total phospholipid weight ratio of 1:2.7 ± 20%, and sphingomyelin and DPPG at a sphingomyelin weight:DPPG weight ratio of phospholipid sphingomyelin of 97:3 ± 20%. 335. The method according to embodiment 334, wherein the formulation comprises ApoA-I and phospholipids at an ApoA-I weight:total phospholipid weight ratio of 1:2.7 ± 10%, and sphingomyelin and DPPG at a sphingomyelin weight:DPPG weight ratio of phospholipid sphingomyelin of 97:3 ± 10%. 336. The method according to embodiment 335, wherein the formulation comprises ApoA-I and phospholipids at an ApoA-I weight:total phospholipid weight ratio of 1:2.7, and sphingomyelin and DPPG at a sphingomyelin weight:DPPG weight ratio of phospholipid sphingomyelin of 97:3. 337. The method according to any one of embodiments 334 to 336, wherein ApoA-I has the amino acid sequence of amino acids 25 to 267 of SEQ ID NO: 2. 338. The method according to any one of embodiments 334 to 337, wherein ApoA-I is expressed by recombinant methods. 339. The method according to embodiment 338, wherein ApoA-I is produced by mammalian host cells. 340. The method according to embodiment 339, wherein the mammalian host cells are Chinese hamster ovary (CHO) cells. 341. The method according to embodiment 340, wherein the CHO cells are CHO-S cells. 342. The method according to any one of embodiments 339 to 341, wherein ApoA-I is subjected to post-translational processing (e.g., glycosylation) such that ApoA-I has one or more structural features (e.g., glycosylation pattern) different from human ApoA-I purified from human plasma. 343. The method according to any one of embodiments 179 to 342, wherein the formulation comprises natural sphingomyelin. 344. The method according to embodiment 343, wherein the natural sphingomyelin is chicken egg sphingomyelin. 345. The method according to any one of embodiments 179 to 342, wherein the formulation comprises synthetic sphingomyelin. 346. The method according to embodiment 345, wherein the synthetic sphingomyelin is palmitoyl sphingomyelin. 347. The method according to any one of embodiments 179 to 346, wherein the formulation is at least 95% homogeneous. 348. The method according to embodiment 347, wherein the formulation is at least 97% homogeneous. 349. The method according to embodiment 347, wherein the formulation is at least 98% homogeneous. 350. The method according to embodiment 347, wherein the formulation is at least 99% homogeneous. 351. The method according to any one of embodiments 179 to 350, wherein the subject is human. 352. The method according to any one of embodiments 179 to 351, wherein the subject is not receiving mechanical ventilation when CER-001 is first administered.

[0221] Although various specific embodiments have been illustrated and described, it is recognized that various changes may be made without departing from the spirit and scope of the present disclosure(s).

[0222] 9. Incorporation by Reference All published materials, patents, patent applications, and other documents cited in this application are hereby incorporated by reference in their entirety for all purposes to the same extent as if each of the published materials, patents, patent applications, or other documents were individually set forth and incorporated by reference for all purposes.

[0223] Any description of documents, acts, materials, devices, articles, etc. contained in this specification is for the sole purpose of providing the content of this disclosure. None of these matters, either individually or in combination, should be taken as an admission that any of them formed part of the prior art base, or was common general knowledge in the field related to this disclosure, as if any or all of them had existed somewhere prior to the priority date of this application.

Claims

1. an apolipoprotein A-I ("ApoA-I") preparation for use in a method of treating subjects with hemophagocytic lymphohistiocytosis (HLH), dengue hemorrhagic fever, or dengue shock syndrome, ApoA-I, in the form of a lipoprotein complex, and the phospholipids sphingomyelin and 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol) ("DPPG"), The formulation comprises ApoA-I and phospholipids in a ratio of 1:2.7 of ApoA-I weight to total phospholipid weight, and a weight-to-weight ratio of 97:3 of sphingomyelin to DPPG.

2. The ApoA-I formulation for use according to claim 1, wherein the subject has or has the risk of HLH following a non-malignant state.

3. The ApoA-I formulation for use according to claim 2, wherein the non-malignant condition is a viral infection.

4. The ApoA-I preparation for use according to claim 3, wherein the subject has HLH caused by dengue fever infection or is at risk of having HLH.

5. The ApoA-I preparation for use according to any one of claims 1 to 4, wherein the subject has dengue fever or dengue hemorrhagic fever.

6. An ApoA-I preparation for use according to any one of claims 1 to 4, wherein the subject has dengue shock syndrome.

7. The ApoA-I formulation for use according to claim 3, wherein the subject has HLH caused by herpes simplex infection or is at risk of having HLH.

8. The ApoA-I formulation for use according to claim 3, wherein the subject has or is at risk of having HLH caused by Epstein-Barr virus infection.

9. The ApoA-I preparation for use according to claim 2, wherein the non-malignant condition is an autoimmune disease.

10. The ApoA-I preparation for use according to claim 1, wherein the subject has or is at risk of HLH following a malignant condition, and the malignant condition may be leukemia or lymphoma.

11. The ApoA-I formulation for use according to claim 1, wherein the subject has or is at risk of familial HLH.

12. ApoA-I formulation for use according to any one of claims 1 to 4, wherein the subject has HLH.

13. An ApoA-I formulation for use according to any one of claims 1 to 4, wherein the ApoA-I has the amino acid sequence of amino acids 25 to 267 of SEQ ID NO: 2, and / or the ApoA-I is expressed by recombinant DNA.

14. A preparation of apolipoprotein A-I ("ApoA-I") for use in a method of treating a subject having dengue fever, herpes simplex infection, or Epstein-Barr infection, ApoA-I, in the form of a lipoprotein complex, and the phospholipids sphingomyelin and 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol) ("DPPG"), The formulation comprises ApoA-I and phospholipids in a ratio of 1:2.7 of ApoA-I weight to total phospholipid weight, and a weight-to-weight ratio of 97:3 of sphingomyelin to DPPG.

15. The ApoA-I formulation for use according to any one of claims 1 to 4 or 14, further comprising administering one or more corticosteroids to the subject, wherein the one or more corticosteroids optionally include methylprednisolone, dexamethasone, or a combination thereof.

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