Lipid-binding protein molecular therapy

Abstract

A method for treating a subject having or at risk of one or more pathological conditions, such as sepsis (e.g., septic shock), using a lipid binding protein molecule.

Description

[Technical Field]

[0001] 1. CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Application Nos. 63 / 479,912, filed January 13, 2023, 63 / 488,835, filed March 7, 2023, and 63 / 594,680, filed October 31, 2023, the contents of each of which are incorporated herein by reference in their entirety.

[0002] 2. Sequence Listing This application contains a Sequence Listing that has been submitted electronically in XML format and is incorporated herein by reference in its entirety. The XML copy, created on December 18, 2023, is designated CRN-051WO_SL.xml and is 4322 bytes in size. [Background technology]

[0003] 3.Background technology Current treatments for sepsis and its sequelae are inadequate. Therefore, new treatments for these conditions are needed. Summary of the Invention

[0004] 4. Summary of the Invention The present disclosure provides methods for treating subjects with or at risk of various pathological conditions. In various embodiments, the pathological conditions are Gram-positive bacterial infection, Gram-negative bacterial infection, viral infection, such as SARS-CoV-2 (COVID-19) infection or influenza virus infection, acute myocardial infarction (AMI), Alzheimer's disease, chronic inflammatory bowel disease (IBD), cardiovascular disease (CVD), stroke, transient ischemic attack, cytokine release syndrome (CRS, cytokine storm), transplanted organ, such as transplanted heart rejection, ischemia-reperfusion-induced tissue injury, postoperative inflammation, psoriasis, sepsis (e.g., septic shock), and the subject has abnormal levels of at least two of TNFα, IL-6, IL-8, TREM-1, and kynurenine pathway biomarkers, for example, abnormal levels of TREM- 1, and sepsis with abnormal levels of at least one of quinolinic acid, kynurenic acid, kynurenine, tryptophan, and kynurenine / tryptophan ratio, including, but not limited to, sepsis, sepsis-induced acute kidney injury (AKI), hypoalbuminemia, attention deficit / hyperactivity disorder (ADHD), central nervous system (CNS) disease, COVID-19 cognitive decline, depression or major depressive disorder, epilepsy, HIV-associated neurocognitive disorder, Huntington's disease, inflammatory bowel disease (IBD), long-term cognitive decline ("brain fog"), such as can develop after sepsis, death or neurological deficit after cardiac arrest, multiple sclerosis (MS), Parkinson's disease, schizophrenia, or vascular endothelial dysfunction.

[0005] The disclosed methods include treating a subject with a lipid-binding protein molecule, e.g., an apolipoprotein, e.g., ApoA-I or an apolipoprotein mimetic. The lipid-binding protein molecule can be administered in high doses, typically a collection of two or more individual doses administered over one or more days, particularly when the symptoms are acute, such as sepsis. The high dose is typically higher than the dose used to treat chronic conditions, e.g., familial hypercholesterolemia. For the treatment of acute conditions, the high dose is typically administered over a relatively short period, e.g., over a period of one day to two weeks or one day to three weeks, and typically includes multiple administrations of the lipid-binding protein molecule, e.g., 2 to 20 individual doses. The individual doses can be separated by less than one day (e.g., two administrations per day) or by one or more days (e.g., one administration per day).

[0006] In some embodiments of the disclosed method, the lipid-binding protein molecule is a component of a lipid-binding protein-based complex. The lipid-binding protein-based complex can include an amphipathic molecule such as a lipid, for example, sphingomyelin and / or a negatively charged lipid. An exemplary lipid-binding protein-based complex that can be used in the disclosed method is CER-001. CER-001 is a negatively charged lipoprotein complex that includes recombinant human ApoA-I, sphingomyelin (SM), and 1,2-dihexadecanoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (dipalmitoylphosphatidyl-glycerol; DPPG).

[0007] As reported in Section 7 of this disclosure, CER-001 therapy has broad pleiotropic effects, as observed in an animal model of LPS-induced acute kidney injury and in clinical trials in septic human patients at high risk for acute kidney injury (AKI). In these studies, CER-001 therapy resulted in, among other things, a reduction in the levels of various inflammatory cytokines, such as IL-6, and in the levels of kynurenine pathway markers. The markers studied are associated with various pathologies in addition to sepsis. Without being bound by theory, it is believed that the pleiotropic effects of CER-001 may extend beyond septic patients to provide clinical benefit to subjects with other pathologies, such as those described herein.

[0008] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of a Gram-positive bacterial infection, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0009] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of a gram-negative bacterial infection, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0010] In one aspect, the present disclosure provides a method of treating a subject having or at risk of a viral infection, e.g., a SARS-CoV-2 (COVID-19) infection or an influenza virus infection, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0011] In one aspect, the present disclosure provides a method of treating a subject having or at risk of acute myocardial infarction (AMI), comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0012] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of Alzheimer's disease, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0013] In one aspect, the present disclosure provides a method of treating a subject having or at risk of chronic inflammatory bowel disease (IBD), comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0014] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of cardiovascular disease (CVD), comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0015] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of having a stroke, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0016] In one aspect, the present disclosure provides a method of treating a subject having or at risk of having a transient ischemic attack, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0017] In one aspect, the present disclosure provides a method of treating a subject having or at risk of cytokine release syndrome (CRS, cytokine storm), comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0018] In one aspect, the present disclosure provides a method of treating a subject having or at risk of transplanted organ, e.g., heart transplant, rejection, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0019] In one aspect, the present disclosure provides a method of treating a subject having or at risk of ischemia-reperfusion-induced tissue injury, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0020] In one aspect, the present disclosure provides a method of treating a subject having or at risk of postoperative inflammation, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0021] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of psoriasis, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0022] In one aspect, the present disclosure provides a method of treating a subject having or at risk of sepsis (e.g., septic shock), including, but not limited to, sepsis in which the subject has abnormal levels of at least two (e.g., 2, 3, 4, or 5) of TNFα, IL-6, IL-8, TREM-1, and kynurenine pathway biomarkers, such as abnormal levels of TREM-1, and abnormal levels of at least one of quinolinic acid, kynurenic acid, kynurenine, tryptophan, and kynurenine / tryptophan ratio, and the method comprises administering a lipid-binding protein molecule (e.g., ApoA-I) to the subject.

[0023] In one aspect, the present disclosure provides a method of treating a subject having or at risk of sepsis-induced acute kidney injury (AKI), comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0024] In one aspect, the present disclosure provides a method of treating a subject having or at risk of having hypoalbuminemia, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0025] In one aspect, the present disclosure provides a method of treating a subject having or at risk of hypoalbuminemia associated with vitamin deficiency, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0026] In one aspect, the present disclosure provides a method of treating a subject having or at risk of hypoalbuminemia associated with inflammatory bowel disease (IBD), comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0027] In one aspect, the present disclosure provides a method of treating a subject having or at risk of having hypoalbuminemia associated with kidney disease, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0028] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of having hypoalbuminemia associated with an infection, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0029] In one aspect, the present disclosure provides a method of treating a subject having or at risk of stress-related hypoalbuminemia, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0030] In one aspect, the present disclosure provides a method of treating a subject having or at risk of hypoalbuminemia associated with thyroid disease, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0031] In one aspect, the present disclosure provides a method of treating a subject having or at risk of diabetes-associated hypoalbuminemia, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0032] In one aspect, the present disclosure provides a method of treating a subject having or at risk of hypoalbuminemia associated with nephrotic syndrome, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0033] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of lupus-associated hypoalbuminemia, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0034] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of cirrhosis-associated hypoalbuminemia, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0035] In one aspect, the present disclosure provides a method of treating a subject having or at risk of hypoalbuminemia associated with liver disease, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0036] In one aspect, the present disclosure provides a method of treating a subject having or at risk of having hypoalbuminemia associated with heart failure, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0037] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of having hypoalbuminemia associated with malnutrition, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0038] In one aspect, the present disclosure provides a method of treating a subject having or at risk of attention-deficit / hyperactivity disorder (ADHD), comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0039] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of a central nervous system (CNS) disease, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0040] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of COVID-19 cognitive decline, comprising administering to the subject a lipid binding protein molecule (e.g., ApoA-I).

[0041] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of depression or major depressive disorder, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0042] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of epilepsy, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0043] In one aspect, the present disclosure provides a method of treating a subject having or at risk of an HIV-associated neurocognitive disorder, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0044] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of Huntington's disease, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0045] In one aspect, the present disclosure provides a method of treating a subject having or at risk of inflammatory bowel disease (IBD), comprising administering to the subject a lipid binding protein molecule (e.g., ApoA-I).

[0046] In one aspect, the present disclosure provides a method of treating a subject having or at risk of long-term cognitive decline ("brain fog"), such as may develop after sepsis, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0047] In one aspect, the present disclosure provides a method of treating a subject having or at risk of death or neurological deficit following cardiac arrest, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0048] In one aspect, the present disclosure provides a method of treating a subject having or at risk of multiple sclerosis (MS), comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0049] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of Parkinson's disease, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0050] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of schizophrenia, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0051] In one aspect, the present disclosure provides a method of treating a subject having or at risk of having a vascular endothelial disorder, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0052] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of a urinary tract infection, comprising administering to the subject a lipid binding protein molecule (e.g., ApoA-I).

[0053] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of a blood-borne infection, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0054] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of a post-surgical infection, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0055] In one aspect, the present disclosure provides a method of treating a subject having or at risk of gastrointestinal perforation, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0056] In one aspect, the present disclosure provides a method of treating a subject having or at risk of having a perforated duodenal ulcer, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0057] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of having a perforated bowel, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0058] In one aspect, the present disclosure provides a method of treating a subject having or at risk of septic shock (e.g., a subject having sepsis and not yet progressing to the stage of septic shock), comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0059] In one embodiment, the disclosure provides a method of treating a subject having or at risk of septic shock following trauma, eg, abdominal trauma.

[0060] In some embodiments, a subject with septic shock has hypotension (e.g., systolic arterial pressure less than 90 mmHg or mean arterial pressure (MAP) less than 65 mmHg) requiring the use of vasopressors despite intravenous fluid resuscitation.

[0061] In one aspect, the present disclosure provides a method of treating a subject having or at risk of having pneumonia, e.g., hospital-acquired pneumonia, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0062] In one aspect, the present disclosure provides a method of treating a subject having or at risk of pancreatitis, e.g., necrotizing pancreatitis, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0063] In some embodiments, the present disclosure provides dosing regimens for lipid binding protein molecule-based therapies (eg, ApoA-I therapy) for subjects described herein.

[0064] The dosing regimens of the present disclosure typically involve multiple administrations of ApoA-I to a subject (e.g., daily or twice-daily administration). ApoA-I therapy can be continued for a predetermined period of time, for example, one week or less (e.g., 1, 2, 3, 4, 5, 6, or 7 days), or for a period longer than one week (e.g., 2 or 3 weeks). Alternatively, administration of ApoA-I to a subject can be continued until one or more symptoms of the condition are reduced, or until the level (e.g., serum level) of one or more relevant biomarkers is reduced, for example, to normal levels or reduced relative to a baseline measurement taken before the initiation of ApoA-I therapy. For subjects with an infection (e.g., a bacterial or viral infection), therapy can, in some embodiments, be continued until the subject recovers from the infection.

[0065] The dosing regimens of the present disclosure can involve administering to a subject a lipid binding protein molecule (e.g., ApoA-I) according to an initial "induction" regimen, and optionally subsequently administering to the subject a lipid binding protein molecule according to a "consolidation" regimen.

[0066] An induction regimen typically involves administering to a subject multiple doses of a lipid-binding protein molecule (e.g., ApoA-I), for example, 6 doses over 3 days, 8 doses over 4 days, 10 doses over 5 days, 12 doses over 6 days, or 14 doses over 7 days.

[0067] The intensification regimen typically comprises administering one or more doses of a lipid-binding protein molecule (e.g., ApoA-I) to a subject after the final dose of the induction regimen, for example, one or more days after the final dose of the induction regimen. In some embodiments, the first dose of the intensification regimen is administered on the third day after the final dose of the induction regimen. For example, the dosing regimen can comprise administering a lipid-binding protein molecule (e.g., ApoA-I) to a subject on days 1, 2, and 3 according to the induction regimen, and administering a lipid-binding protein molecule to a subject on day 6 according to the intensification regimen. In some embodiments, the intensification regimen comprises two doses of a lipid-binding protein molecule.

[0068] In certain embodiments, the lipid binding protein molecule (eg, ApoA-I) is administered in combination with a standard of care therapy for the disease or condition of interest.

[0069] In certain embodiments, an antihistamine (e.g., dexchlorpheniramine, hydroxyzine, diphenhydramine, cetirizine, fexofenadine, or loratadine) can be administered before administration of a lipid-binding protein molecule (e.g., ApoA-I). Antihistamines can reduce the likelihood of an allergic reaction.

[0070] Further features of exemplary lipid-binding protein molecules and lipid-binding protein-based complexes that can be used in the methods and dosing regimens of the present disclosure are described in Section 6.1 below, and in specific embodiments 382-407 and 678-698.

[0071] Further characteristics of subjects that can be treated according to the methods and dosing regimens of the present disclosure are described in Section 6.2 below, as well as in specific embodiments 1-201, 332-381, 651-672, and 700-707.

[0072] Further features of exemplary dosing regimens of the present disclosure are described below in Section 6.3, as well as in specific embodiments 202-331, 408-636, 673-677, 699, and 709-717.

[0073] Exemplary combination therapies are described below in Section 6.4 and in specific embodiments 637-650. [Brief explanation of the drawings]

[0074] [Figure 1A] FIG. 1 shows quinolinic acid levels for individual endotoxemic pigs ("LPS") and individual endotoxemic pigs treated with one dose of CER-001 at 20 mg / kg ("20 mg") in the study described in Example 1. [Figure 1B] FIG. 1 shows quinolinic acid levels for an individual endotoxemic pig ("LPS") and an individual endotoxemic pig treated with two doses of CER-001 at 20 mg / kg, totaling 40 mg / kg ("40 mg"), in the study described in Example 1. [Figure 1C] 1 shows quinolinic acid levels for all three groups of endotoxemic pigs in the study described in Example 1: LPS, 20 mg, and 40 mg. [Figure 2A] FIG. 1 shows kynurenic acid levels for an individual endotoxemic pig (“LPS”) and an individual endotoxemic pig treated with one dose of CER-001 at 20 mg / kg (“20 mg”) in the study described in Example 1. [Figure 2B] FIG. 1 shows kynurenic acid levels for an individual endotoxemic pig (“LPS”) and an individual endotoxemic pig treated with two doses of CER-001 at 20 mg / kg each (“40 mg”) in the study described in Example 1. [Figure 2C] 1 shows kynurenic acid levels for all three groups of endotoxemic pigs in the study described in Example 1: LPS, 20 mg, and 40 mg. [Figure 3A] 1 shows tryptophan levels for individual endotoxemic pigs ("LPS") and individual endotoxemic pigs treated with one dose of CER-001 at 20 mg / kg ("20 mg") in the study described in Example 1. [Figure 3B] 1 shows tryptophan levels for an individual endotoxemic pig ("LPS") and an individual endotoxemic pig treated with two doses of CER-001 at 20 mg / kg each ("40 mg") in the study described in Example 1. [Figure 3C] 1 shows tryptophan levels for all three groups of endotoxemic pigs in the study described in Example 1: LPS, 20 mg, and 40 mg. [Figure 4A] FIG. 1 shows kynurenine levels for an individual endotoxemic pig (“LPS”) and an individual endotoxemic pig treated with one dose of CER-001 at 20 mg / kg (“20 mg”) in the study described in Example 1. [Figure 4B] FIG. 1 shows kynurenine levels for an individual endotoxemic pig (“LPS”) and an individual endotoxemic pig treated with two doses of CER-001 at 20 mg / kg each (“40 mg”) in the study described in Example 1. [Figure 4C] 1 shows kynurenine levels for all three groups of endotoxemic pigs in the study described in Example 1: LPS, 20 mg, and 40 mg. [Figure 5A] 1 shows the kynurenine / tryptophan ratio for the first cohort of endotoxemic pigs from all three LPS, 20 mg, and 40 mg groups in the study described in Example 1. [Figure 5B] 1 shows the kynurenine / tryptophan ratio for the second cohort of endotoxemic pigs from all three LPS, 20 mg, and 40 mg groups in the study described in Example 1. [Figure 6A]FIG. 1 shows the relative fold gene expression of indoleamine 2,3-dioxygenase 1 (IDO1) in brain tissue as measured by qPCR for cohorts from all three LPS, 20 mg, and 40 mg groups of endotoxemic pigs in the study described in Example 1. [Figure 6B] FIG. 1 shows the relative fold gene expression of aromatic L-amino acid / L-tryptophan decarboxylase (DDC) in brain tissue as measured by qPCR for cohorts from all three LPS, 20 mg, and 40 mg groups of endotoxemic pigs in the study described in Example 1. [Figure 6C] FIG. 1 shows the relative fold gene expression of kynurenine formamidase isoform X1 (AFMID) in brain tissue as measured by qPCR for cohorts from all three LPS, 20 mg, and 40 mg groups of endotoxemic pigs in the study described in Example 1. [Figure 6D] FIG. 1 shows the relative fold gene expression of kynurenine 3-monooxygenase (KMO) in brain tissue as measured by qPCR for cohorts from all three LPS, 20 mg, and 40 mg groups of endotoxemic pigs in the study described in Example 1. [Figure 6E] FIG. 1 shows the relative fold gene expression of kynurenine-oxoglutarate transaminase 3 (KYAT3) in brain tissue as measured by qPCR for cohorts from all three LPS, 20 mg, and 40 mg groups of endotoxemic pigs in the study described in Example 1. [Figure 6F] 1 shows the relative fold gene expression of interleukin-6 (IL-6) in brain tissue as measured by qPCR for cohorts from all three LPS, 20 mg, and 40 mg groups of endotoxemic pigs in the study described in Example 1. [Figure 7] 1 shows a schematic diagram of the clinical study of Example 2. [Figure 8A]1 shows the lipopolysaccharide (LPS) change from baseline for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 8B] 1 shows the LPS change from baseline for each of Groups A to D in the clinical study of Example 2. [Figure 8C] 1 shows LPS changes as a percentage of peak for the standard of care (SOC) group (Group A) and the three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 8D] 1 shows LPS changes as a percentage of peak for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2, separated by whether subjects were enrolled from the center's ICU or nephrology department. [Figure 8E] 1 shows the lipopolysaccharide (LPS) change from baseline for each subject in the standard of care (SOC) group and the three experimental groups (CER-001) in the clinical study of Example 2. [Figure 8F] 1 shows the lipopolysaccharide (LPS) change from baseline for each subject in the standard of care (SOC) group and each of the three experimental groups (CER-001) in the clinical study of Example 2. [Figure 9A] 1 shows the endotoxin activity assay (EAA) change from baseline for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 9B] 1 shows the EAA change from baseline for each of Groups A to D in the clinical study of Example 2. [Figure 9C] 1 shows EAA changes as percentage of peak for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 9D] 1 shows EAA changes as a percentage of peak for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2, separated by whether subjects were enrolled from the center's ICU or nephrology department. [Figure 9E]1 shows the endotoxin activity assay (EAA) change from baseline for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 10A] 1 shows the change in TNF-alpha from baseline for the standard of care (SOC) group (Group A) and three experimental groups (Groups BD) in the clinical study of Example 2. [Figure 10B] 1 shows the change in TNF-alpha from baseline for each of Groups AD in the clinical study of Example 2. [Figure 10C] 1 shows the TNF-alpha change as a percentage of peak for the standard of care (SOC) group (Group A) and the three experimental groups (Groups BD) in the clinical study of Example 2. [Figure 10D] 1 shows the TNF-alpha change as a percentage of peak for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2, separated by whether subjects were enrolled from the center's ICU or nephrology department. [Figure 10E] 1 shows the change in TNF-alpha from baseline for each subject in the standard of care (SOC) group (Group A) and the three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 10F] 1 shows the change in TNF-alpha from baseline for each subject in the standard of care (SOC) group (Group A) and each of the three experimental groups (Groups BD) in the clinical study of Example 2. [Figure 11A] 1 shows the MCP-1 change from baseline for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 11B] 1 shows the change in MCP-1 from baseline for each of Groups A to D in the clinical study of Example 2. [Figure 11C] 1 shows the MCP-1 change as a percentage of peak for the standard of care (SOC) group (Group A) and the three experimental groups (Groups BD) in the clinical study of Example 2. [Figure 11D]1 shows MCP-1 changes as a percentage of peak for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2, separated by whether subjects were enrolled from the ICU or nephrology department of the center. [Figure 11E] 1 shows the change in MCP-1 from baseline for each subject in the standard of care (SOC) group (Group A) and the three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 11F] 1 shows the MCP-1 change from baseline for each subject in the standard of care (SOC) group (Group A) and each of the three experimental groups (Groups BD) in the clinical study of Example 2. [Figure 12A] 1 shows the IL-6 change from baseline for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 12B] 1 shows the change in IL-6 from baseline for each of Groups A to D in the clinical study of Example 2. [Figure 12C] IL-6 change as a percentage of peak for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 12D] IL-6 change as a percentage of peak for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2, separated by whether subjects were enrolled from the center's ICU or nephrology department. [Figure 12E] 1 shows the change in IL-6 from baseline for each subject in the standard of care (SOC) group (Group A) and the three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 12F] 1 shows the change in IL-6 from baseline for each subject in the standard of care (SOC) group (Group A) and each of the three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 13A] 1 shows the change in IL-8 from baseline for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 13B] 1 shows the change in IL-8 from baseline for each of Groups A to D in the clinical study of Example 2. [Figure 13C] IL-8 change as a percentage of peak for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2 are shown. [Figure 13D] IL-8 change as a percentage of peak for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2, separated by whether subjects were enrolled from the center's ICU or nephrology department. [Figure 13E] 1 shows the change in IL-8 from baseline for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 13F] 1 shows the change in IL-8 from baseline for the standard of care (SOC) group (Group A) and each of the three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 14A] 1 shows the IL-10 change from baseline for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 14B] 1 shows the change in IL-10 from baseline for each of Groups A to D in the clinical study of Example 2. [Figure 14C] IL-10 change as a percentage of peak for the standard of care (SOC) group (Group A) and the three experimental groups (Groups B-D) in the clinical study of Example 2 are shown. [Figure 14D] IL-10 change as a percentage of peak for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2, separated by whether subjects were enrolled from the center's ICU or nephrology department. [Figure 15A] 1 shows the TREM-1 change from baseline for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 15B]1 shows the TREM-1 change from baseline for each of Groups A to D in the clinical study of Example 2. [Figure 15C] 1 shows TREM-1 changes as a percentage of peak for the standard of care (SOC) group (Group A) and the three experimental groups (Groups BD) in the clinical study of Example 2. [Figure 15D] 1 shows TREM-1 changes as a percentage of peak for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2, separated by whether subjects were enrolled from the center's ICU or nephrology department. [Figure 15E] 1 shows the TREM-1 change from baseline for each subject in the standard of care (SOC) group (Group A) and the three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 15F] 1 shows the TREM-1 change from baseline for each subject in the standard of care (SOC) group (Group A) and each of the three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 16A] 1 shows VCAM changes from baseline for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 16B] 1 shows the VCAM change from baseline for each of Groups A to D in the clinical study of Example 2. [Figure 16C] 1 shows VCAM changes as a percentage of peak for the standard of care (SOC) group (Group A) and three experimental groups (Groups BD) in the clinical study of Example 2. [Figure 16D] 1 shows VCAM changes as a percentage of peak for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2, separated by whether subjects were enrolled from the center's ICU or nephrology department. [Figure 16E] 1 shows the VCAM change from baseline for each subject in the standard of care (SOC) group (Group A) and the three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 16F]1 shows the VCAM change from baseline for each subject in the standard of care (SOC) group (Group A) and each of the three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 17A] 1 shows the ICAM change from baseline for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 17B] 1 shows the change in ICAM from baseline for each of Groups A to D in the clinical study of Example 2. [Figure 17C] 1 shows ICAM changes as a percentage of peak for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 17D] 1 shows ICAM changes as a percentage of peak for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2, separated by whether subjects were enrolled from the center's ICU or nephrology department. [Figure 17E] 1 shows the ICAM change from baseline for each subject in the standard of care (SOC) group (Group A) and the three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 17F] 1 shows the ICAM change from baseline for each subject in the standard of care (SOC) group (Group A) and each of the three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 18A] 1 shows ferritin changes from baseline for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 18B] 1 shows the ferritin change from baseline for each of Groups A to D in the clinical study of Example 2. [Figure 18C] 1 shows ferritin change as a percentage of peak for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 18D]1 shows ferritin change as a percentage of peak for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2, separated by whether subjects were enrolled from the center's ICU or nephrology department. [Figure 19A] 1 shows the change in white blood cell count from baseline for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 19B] 1 shows the change in white blood cell count from baseline for each of Groups A to D in the clinical study of Example 2. [Figure 19C] 1 shows the white blood cell count change as a percentage of peak for the standard of care (SOC) group (Group A) and the three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 19D] 1 shows the white blood cell count change as a percentage of peak for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2, separated by whether subjects were enrolled from the center's ICU or nephrology department. [Figure 20A] 1 shows the C-reactive protein (CRP) change from baseline for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 20B] 1 shows the change in CRP from baseline for each of Groups A to D in the clinical study of Example 2. [Figure 20C] 1 shows the CRP change as a percentage of peak for the standard of care (SOC) group (Group A) and the three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 20D] 1 shows the CRP change as a percentage of peak for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2, separated by whether subjects were enrolled from the center's ICU or nephrology department. [Figure 20E]1 shows the C-reactive protein (CRP) change from baseline for each subject in the standard of care (SOC) group (Group A) and the three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 20F] 1 shows the C-reactive protein (CRP) change from baseline for each subject in the standard of care (SOC) group (Group A) and each of the three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 21A] 1 shows the KIM-1 change from baseline for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 21B] 1 shows the change in KIM-1 from baseline for each of Groups A to D in the clinical study of Example 2. [Figure 21C] 1 shows KIM-1 changes as a percentage of peak for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 21D] 1 shows KIM-1 changes as a percentage of peak for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2, separated by whether subjects were enrolled from the ICU or nephrology department of the center. [Figure 22A] 1 shows the serum albumin change from baseline for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 22B] 1 shows the serum albumin change from baseline for each of Groups A to D in the clinical study of Example 2. [Figure 22C] 1 shows serum albumin change as a percentage of peak for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 22D] 1 shows serum albumin change as a percentage of peak for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2, separated by whether subjects were enrolled from the center's ICU or nephrology department. [Figure 22E] 1 shows the serum albumin change from baseline for each subject in the standard of care (SOC) group (Group A) and the three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 23A] 1 shows the serum creatinine change from baseline for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 23B] 1 shows the serum creatinine change from baseline for each of Groups A to D in the clinical study of Example 2. [Figure 23C] 1 shows serum creatinine change as a percentage of peak for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 23D] 1 shows the serum creatinine change as a percentage of peak for the standard of care (SOC) group (Group A) and three experimental groups (Groups B-D) in the clinical study of Example 2, separated by whether subjects were enrolled from the center's ICU or nephrology department. [Figure 23E] 1 shows the AUC (mean±SEM) for serum creatinine for the standard of care (SOC) group (Group A) and the three experimental groups (Groups BD) in the clinical study of Example 2. [Figure 23F] 1 shows the AUC (95% confidence interval) for serum creatinine for the standard of care (SOC) group (Group A) and the three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 24A] 1 shows the estimated glomerular filtration rate (eGFR) change from baseline for all subjects in the standard of care (SOC) group (Group A) and the three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 24B] 1 shows the change in eGFR from baseline for all subjects in each of Groups A to D in the clinical study of Example 2. [Figure 24C] 1 shows the change in eGFR from baseline for only subjects with AKI who participated in the clinical study of Example 2 in the standard of care (SOC) group (Group A) and the three experimental groups (Groups B-D). [Figure 24D] Figure 24C shows the eGFR change from baseline for the same subjects as in Figure 24C. [Figure 24E] 1 shows the eGFR change as a percentage of peak for all subjects in the standard of care (SOC) group (Group A) and population groups BD in the clinical study of Example 2. [Figure 24F] 1 shows the eGFR change as a percentage of peak for only subjects with AKI who entered the study in the standard of care (SOC) group (Group A) and population groups BD in the clinical study of Example 2. FIG. [Figure 25] 1 shows the P / F change from baseline for all subjects in the standard of care (SOC) group (Group A) and the three experimental groups (Groups B-D) in the clinical study of Example 2. [Figure 26] 1 shows the survival percentage for all subjects after days in ICU for the standard of care group (Group A, "SOC") and cohort groups BD ("CER-001") in the clinical study of Example 2. [Figure 27A] 1 shows the 30-day survival rates for all subjects for the standard of care group (Group A, "SOC") and aggregation groups BD ("CER-001") in the clinical study of Example 2. [Figure 27B] 1 shows the 30-day survival rate for all subjects enrolled in the study from the center's ICU for the standard of care group (Group A, "SOC") and aggregation groups B through D ("CER-001") in the clinical study of Example 2. [Figure 28A] 1 shows the progression of AKI staging for the standard of care group (Group A, "SOC") in the clinical study of Example 2. [Figure 28B] 1 shows the progression of AKI staging for population groups B to D ("CER-001") in the clinical study of Example 2. [Figure 29] 1 shows the number of days on mechanical ventilation for all subjects enrolled in the study from the center's ICU for the standard of care group (Group A, "SOC") and cluster groups B through D ("CER-001") in the clinical study of Example 2. [Figure 30] 1 shows the number of days on vasoconstrictor therapy for all subjects enrolled in the study from the center's ICU for the standard of care group (Group A, "SOC") and aggregate groups B through D ("CER-001") in the clinical study of Example 2. [Figure 31A] 1 shows the number of days on dialysis for all subjects enrolled in the study from the center's ICU for the standard of care group (Group A, "SOC") and aggregation groups B through D ("CER-001") in the clinical study of Example 2. [Figure 31B] 1 shows the number of days on dialysis for all subjects who participated in the standard of care group (Group A, "SOC") and population groups B through D ("CER-001") in the clinical study of Example 2. [Figure 32] 1 shows the organ support-free survival in days for all subjects enrolled in the study from the center's ICU for the standard of care group (Group A, "SOC") and aggregate groups B through D ("CER-001") in the clinical study of Example 2. [Figure 33A] 1 shows the change in daily mean arterial pressure (MAP) for all subjects enrolled in the study from the center's ICU for the standard of care group (Group A, "SOC") and aggregation groups B through D ("CER-001") in the clinical study of Example 2. [Figure 33B] 1 shows the change in daily mean arterial pressure (MAP) for each subject enrolled in the study from the center's ICU for the standard of care group (Group A, "SOC") and cluster groups B through D ("CER-001") in the clinical study of Example 2. [Figure 34] 1 shows the change in daily mean heart rate (HR) for all subjects enrolled in the study from the center's ICU for the standard of care group (Group A, "SOC") and cluster groups B through D ("CER-001") in the clinical study of Example 2. [Figure 35] 1 shows the change in the mean daily P / F ratio for all subjects enrolled in the study from the center's ICU for the standard of care group (Group A, "SOC") and collective groups B through D ("CER-001") in the clinical study of Example 2. [Figure 36]1 shows survival curves of pigs upon challenge with LPS and injection with CER-001, as described in Example 3. [Figure 37] 1 shows serum VCAM levels in pigs upon challenge with LPS and injection of CER-001, as described in Example 3. [Figure 38] 1 shows serum ICAM levels in pigs upon challenge with LPS and injection of CER-001, as described in Example 3. [Figure 39] 1 shows serum TNF-α levels in pigs upon challenge with LPS and injection of CER-001, as described in Example 3. [Figure 40] 1 shows serum MCP-1 levels in pigs upon challenge with LPS and injection with CER-001, as described in Example 3. [Figure 41] 1 shows serum 11-6 levels in pigs upon challenge with LPS and injection of CER-001, as described in Example 3. [Figure 42] 1 shows systemic classical pathway complement activation in pigs upon challenge with LPS and injection of CER-001, as described in Example 3. [Figure 43] 1 shows systemic alternative pathway complement activation in pigs upon challenge with LPS and injection of CER-001, as described in Example 3. [Figure 44] 1 shows systemic lectin pathway complement activation in pigs upon challenge with LPS and injection of CER-001, as described in Example 3. [Figure 45A] 1 shows representative hematoxylin and eosin (HE) staining of pig liver tissue upon challenge with LPS, as described in Example 3. [Figure 45B] 1 shows representative HE staining of pig liver tissue upon challenge with LPS and CER-001 treatment (20 mg / kg), as described in Example 3. [Figure 45C] 1 shows representative HE staining of pig liver tissue upon challenge with LPS and treatment with CER-001 (20 mg / kg x 2), as described in Example 3. [Figure 45D]1 shows liver injury quantified from images of stained pig livers upon challenge with LPS and injection of CER-001, as described in Example 3. [Figure 45E] 1 shows serum levels of ALT enzyme in pigs upon challenge with LPS and injection of CER-001, as described in Example 3. [Figure 46A] 1 shows representative HE staining of pig kidney tissue upon challenge with LPS, as described in Example 3. [Figure 46B] 1 shows representative HE staining of pig kidney tissue upon challenge with LPS and CER-001 treatment (20 mg / kg), as described in Example 3. [Figure 46C] 1 shows representative HE staining of pig kidney tissue upon challenge with LPS and treatment with CER-001 (20 mg / kg x 2), as described in Example 3. [Figure 46D] 1 shows tubular pathology scores quantified from images of stained kidneys of pigs upon challenge with LPS and injection of CER-001, as described in Example 3. [Figure 46E] 1 shows glomerular pathology scores quantified from images of stained kidneys of pigs upon challenge with LPS and injection of CER-001, as described in Example 3. [Figure 47] 1 shows serum levels of creatinine in pigs upon challenge with LPS and injection of CER-001, as described in Example 3. [Figure 48] 1 shows urine output in pigs upon challenge with LPS and injection of CER-001, as described in Example 3. [Figure 49A] 1 shows serum cystatin C levels in pigs upon challenge with LPS and injection of CER-001, as described in Example 3. [Figure 49B] 1 shows urinary cystatin C levels in pigs upon challenge with LPS and injection of CER-001, as described in Example 3. [Figure 50A] 1 shows serum KIM-1 levels in pigs upon challenge with LPS and injection of CER-001, as described in Example 3. [Figure 50B]1 shows urinary KIM-1 levels in pigs upon challenge with LPS and injection of CER-001, as described in Example 3. [Figure 51] 1 shows serum LPS levels in pigs upon challenge with LPS and injection of CER-001, as described in Example 3. [Figure 52A] 1 shows a Western blot of LPS and β-actin protein expression in pig liver upon challenge with LPS and CER-001 injection, as described in Example 3. [Figure 52B] 1 shows densitometric analysis of LPS and β-actin protein expression in pig liver upon challenge with LPS and CER-001 injection, as described in Example 3. [Figure 53] 1 shows endotoxin levels in pig bile upon challenge with LPS and injection of CER-001, as described in Example 3. [Figure 54] 1 shows serum levels of human ApoA-I in pigs upon challenge with LPS and injection of CER-001, as described in Example 3. [Figure 55] 1 shows levels of human ApoA-I in pig bile upon challenge with LPS and injection of CER-001, as described in Example 3. [Figure 56A] 1 shows the mean ApoA-I levels for the control and population study groups in the clinical study of Example 2. [Figure 56B] 1 shows the ApoA-I levels for each subject separated by study group in the clinical study of Example 2. [Figure 56C] 1 shows the change from baseline in ApoA-I levels for each subject in the clinical study of Example 2. [Figure 56D] 1 shows the change from baseline in ApoA-I levels for each subject divided by study group in the clinical study of Example 2. [Figure 57A] 1 shows the change from baseline in aspartate transaminase (AST) levels for each subject in the clinical study of Example 2. [Figure 57B]1 shows the change from baseline in alanine transaminase (ALT) levels for each subject in the clinical study of Example 2. [Figure 58] 1 shows the results of an MTT cell viability assay on cultured endothelial cells upon challenge with LPS and injection of CER-001, as described in Example 4. [Figure 59] 1 summarizes endothelial nitric oxide synthase (eNOS)-based (eNOS (phospho-S1177)) FACS results for cultured endothelial cells upon challenge with LPS and CER-001 infusion, as described in Example 4. [Figure 60] 1 shows eNOS (phospho-S1177)-based FACS results for cultured endothelial cells upon challenge with LPS and CER-001 infusion in one representative of three independent experiments, compared to basal and VEFG (positive control) cells, as described in Example 4. [Figure 61] 1 shows the results of an MTT cell viability assay on PBMCs from healthy donors upon challenge with LPS and CER-001 infusion, as described in Example 4. [Figure 62] 1 shows TNF-α synthesis for PBMCs from healthy donors upon challenge with LPS and CER-001 infusion, as described in Example 4. [Figure 63] 1 shows CD14-based FACS results for PBMCs from healthy donors upon challenge with LPS and infusion of CER-001 in one representative of three independent experiments, as described in Example 4. [Figure 64] 1 summarizes CD14-based FACS results for PBMCs from healthy donors upon challenge with LPS and CER-001 infusion, as described in Example 4. [Figure 65] 1 shows the number of days to ICU discharge for all subjects enrolled in the study from the center's ICU for the standard of care group (Group A, "SOC") and aggregation groups B through D ("CER-001") in the clinical study of Example 2. [Figure 66]1 shows the change in serum quinolinic acid (QA) levels from baseline (Day 1) for subjects in the standard of care (SOC) group (Group A) and population groups BD in the clinical study of Example 2. [Figure 67] 1 shows the change in serum kynurenine / tryptophan ratio (Kyn / Trp) levels from baseline (Day 1) for subjects in the standard of care (SOC) group (Group A) and population groups BD in the clinical study of Example 2. [Figure 68] 1 shows the change in serum serotonin levels from baseline (Day 1) for subjects in the standard of care (SOC) group (Group A) and population groups BD in the clinical study of Example 2. [Figure 69] 1 shows the overall trial design for Example 5. [Figure 70] 1 shows study participation for individual subjects in Example 5. DETAILED DESCRIPTION OF THE INVENTION

[0075] 6. MODE FOR CARRYING OUT THE INVENTION The present disclosure provides methods for treating subjects with or at risk of various pathologies with lipid-binding protein molecules. In various embodiments, the pathologies include gram-positive bacterial infection, gram-negative bacterial infection, viral infection, such as SARS-CoV-2 (COVID-19) infection or influenza virus infection, acute myocardial infarction (AMI), Alzheimer's disease, chronic inflammatory bowel disease (IBD), cardiovascular disease (CVD), stroke, transient ischemic attack, cytokine release syndrome (CRS, cytokine storm), transplanted organ (e.g., cardiac transplant rejection), ischemia-reperfusion-induced tissue injury, postoperative inflammation, psoriasis, and sepsis.

[0023] The present invention relates to a pulmonary edema (PED) characterized by sepsis, sepsis-induced acute kidney injury (AKI), hypoalbuminemia, hypovitaminosis (hypovitaminosis), ... The following conditions may be present: hypoalbuminemia associated with insulin deficiency, hypoalbuminemia associated with inflammatory bowel disease (IBD), hypoalbuminemia associated with kidney disease, hypoalbuminemia associated with infection, hypoalbuminemia associated with stress, hypoalbuminemia associated with thyroid disease, hypoalbuminemia associated with diabetes, hypoalbuminemia associated with nephrotic syndrome, hypoalbuminemia associated with lupus, hypoalbuminemia associated with cirrhosis, hypoalbuminemia associated with liver disease, hypoalbuminemia associated with heart failure, hypoalbuminemia associated with malnutrition, attention deficit / hyperactivity disorder (ADHD), central nervous system (CNS) disease, COVID-19 cognitive decline, depression or major depressive disorder, epilepsy, HIV-associated neurocognitive disorder, Huntington's disease, inflammatory bowel disease (IBD), long-term cognitive decline ("brain fog"), such as can develop after sepsis, death or neurological deficit after cardiac arrest, multiple sclerosis (MS), Parkinson's disease, schizophrenia, or vascular endothelial dysfunction.

[0076] In some embodiments, the methods involve administering a high dose of a lipid binding protein molecule.

[0077] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of a Gram-positive bacterial infection, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0078] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of a gram-negative bacterial infection, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0079] In one aspect, the present disclosure provides a method of treating a subject having or at risk of a viral infection, e.g., a SARS-CoV-2 (COVID-19) infection or an influenza virus infection, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0080] In one aspect, the present disclosure provides a method of treating a subject having or at risk of acute myocardial infarction (AMI), comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0081] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of Alzheimer's disease, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0082] In one aspect, the present disclosure provides a method of treating a subject having or at risk of chronic inflammatory bowel disease (IBD), comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0083] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of cardiovascular disease (CVD), comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0084] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of having a stroke, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0085] In one aspect, the present disclosure provides a method of treating a subject having or at risk of having a transient ischemic attack, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0086] In one aspect, the present disclosure provides a method of treating a subject having or at risk of cytokine release syndrome (CRS, cytokine storm), comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0087] In one aspect, the present disclosure provides a method of treating a subject having or at risk of transplanted organ, e.g., heart transplant, rejection, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0088] In one aspect, the present disclosure provides a method of treating a subject having or at risk of ischemia-reperfusion-induced tissue injury, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0089] In one aspect, the present disclosure provides a method of treating a subject having or at risk of postoperative inflammation, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0090] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of psoriasis, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0091] In one aspect, the present disclosure provides a method of treating a subject having or at risk of sepsis (e.g., septic shock), including, but not limited to, sepsis in which the subject has abnormal levels of at least two of TNFα, IL-6, IL-8, TREM-1, and kynurenine pathway biomarkers, for example, abnormal levels of TREM-1, and abnormal levels of at least one of quinolinic acid, kynurenic acid, kynurenine, tryptophan, and kynurenine / tryptophan ratio, and the method comprises administering a lipid-binding protein molecule (e.g., ApoA-I) to the subject.

[0092] In one aspect, the present disclosure provides a method of treating a subject having or at risk of sepsis-induced acute kidney injury (AKI), comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0093] In one aspect, the present disclosure provides a method of treating a subject having or at risk of having hypoalbuminemia, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0094] In one aspect, the present disclosure provides a method of treating a subject having or at risk of hypoalbuminemia associated with vitamin deficiency, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0095] In one aspect, the present disclosure provides a method of treating a subject having or at risk of hypoalbuminemia associated with inflammatory bowel disease (IBD), comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0096] In one aspect, the present disclosure provides a method of treating a subject having or at risk of having hypoalbuminemia associated with kidney disease, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0097] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of having hypoalbuminemia associated with an infection, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0098] In one aspect, the present disclosure provides a method of treating a subject having or at risk of stress-related hypoalbuminemia, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0099] In one aspect, the present disclosure provides a method of treating a subject having or at risk of hypoalbuminemia associated with thyroid disease, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0100] In one aspect, the present disclosure provides a method of treating a subject having or at risk of diabetes-associated hypoalbuminemia, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0101] In one aspect, the present disclosure provides a method of treating a subject having or at risk of hypoalbuminemia associated with nephrotic syndrome, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0102] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of lupus-associated hypoalbuminemia, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0103] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of cirrhosis-associated hypoalbuminemia, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0104] In one aspect, the present disclosure provides a method of treating a subject having or at risk of hypoalbuminemia associated with liver disease, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0105] In one aspect, the present disclosure provides a method of treating a subject having or at risk of having hypoalbuminemia associated with heart failure, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0106] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of having hypoalbuminemia associated with malnutrition, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0107] In one aspect, the present disclosure provides a method of treating a subject having or at risk of attention-deficit / hyperactivity disorder (ADHD), comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0108] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of a central nervous system (CNS) disease, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0109] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of COVID-19 cognitive decline, comprising administering to the subject a lipid binding protein molecule (e.g., ApoA-I).

[0110] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of depression or major depressive disorder, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0111] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of epilepsy, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0112] In one aspect, the present disclosure provides a method of treating a subject having or at risk of an HIV-associated neurocognitive disorder, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0113] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of Huntington's disease, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0114] In one aspect, the present disclosure provides a method of treating a subject having or at risk of inflammatory bowel disease (IBD), comprising administering to the subject a lipid binding protein molecule (e.g., ApoA-I).

[0115] In one aspect, the present disclosure provides a method of treating a subject having or at risk of long-term cognitive decline ("brain fog"), such as may develop after sepsis, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0116] In one aspect, the present disclosure provides a method of treating a subject having or at risk of death or neurological deficit following cardiac arrest, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0117] In one aspect, the present disclosure provides a method of treating a subject having or at risk of multiple sclerosis (MS), comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0118] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of Parkinson's disease, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0119] In one embodiment, the present disclosure provides a method of treating a subject having or at risk of schizophrenia, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0120] In one aspect, the present disclosure provides a method of treating a subject having or at risk of having a vascular endothelial disorder, comprising administering to the subject a lipid-binding protein molecule (e.g., ApoA-I).

[0121] In some embodiments, the condition is associated with abnormal levels of one or more of TREM-1, albumin, interleukin-10 (IL-10), kynurenine pathway biomarkers (e.g., tryptophan, serotonin, formylkynurenine, kynurenine, kynurenic acid, 2-amino-3-carboxymuconic acid-semialdehyde, 3-hydroxykynurenine, xanthurenic acid, anthralinic acid, 3-hydroxyanthrarinic acid, quinolinic acid, picolinic acid, quinaldic acid, or the kynurenine / tryptophan ratio), TNF-α, MCP-1, IL-6, IL-8, IL-10, VCAM-1, or ICAM-1.

[0122] In some embodiments, the condition is associated with abnormal levels of 2-amino-3-carboxymuconic acid-semialdehyde.

[0123] In some embodiments, the condition is associated with abnormal levels of 3-hydroxyanthranilic acid.

[0124] In some embodiments, the condition is associated with abnormal levels of 3-hydroxykynurenine.

[0125] In some embodiments, the condition is associated with abnormal levels of albumin.

[0126] In some embodiments, the condition is associated with abnormal levels of anthranilic acid.

[0127] In some embodiments, the condition is associated with abnormal levels of formylkynurenine.

[0128] In some embodiments, the condition is associated with abnormal levels of ICAM-1.

[0129] In some embodiments, the condition is associated with abnormal levels of IL-6.

[0130] In some embodiments, the condition is associated with abnormal levels of IL-8.

[0131] In some embodiments, the condition is associated with abnormal levels of kynurenic acid.

[0132] In some embodiments, the condition is associated with abnormal levels of kynurenine.

[0133] In some embodiments, the condition is associated with an abnormal level of the kynurenine / tryptophan ratio.

[0134] In some embodiments, the condition is associated with abnormal levels of MCP-1.

[0135] In some embodiments, the condition is associated with abnormal levels of picolinic acid.

[0136] In some embodiments, the condition is associated with abnormal levels of quinaldic acid.

[0137] In some embodiments, the condition is associated with abnormal levels of quinolinic acid.

[0138] In some embodiments, the condition is associated with abnormal levels of serotonin.

[0139] In some embodiments, the condition is associated with abnormal levels of TNFα.

[0140] In some embodiments, the condition is associated with abnormal levels of TREM-1.

[0141] In some embodiments, the condition is associated with abnormal levels of tryptophan.

[0142] In some embodiments, the condition is associated with abnormal levels of VCAM-1.

[0143] In some embodiments, the condition is associated with abnormal levels of xanthurenic acid.

[0144] In some embodiments, the lipid-binding protein molecule is provided as a component of lipid-binding protein-based complex.In some embodiments, the lipid-binding protein-based complex is Apomer, Cargomer, HDL-based complex or HDL-mimetic-based complex.In specific embodiments, the lipid-binding protein-based complex is CER-001.

[0145] Exemplary characteristics of lipid-binding protein molecules and lipid-binding protein-based complexes comprising lipid-binding protein molecules that can be used in the methods and compositions of the present disclosure are described in Section 6.1. Exemplary subject populations that can be treated by the methods and compositions of the present disclosure are described in Section 6.2.

[0146] In some embodiments, the method of the present disclosure comprises administering a lipid-binding protein molecule (e.g., ApoA-I) to a subject in two phases. First, the lipid-binding protein molecule (e.g., ApoA-I) is administered in an initial, high-intensity "induction" regimen. The induction regimen is followed by a less intense "consolidation" regimen. Alternatively, the lipid-binding protein molecule (e.g., ApoA-I) can be administered to a subject in a single phase, for example, according to a dosing regimen corresponding to the dose and dosing frequency of the induction or consolidation regimen described herein.

[0147] Induction regimens that can be used in the methods of the present disclosure are described in Section 6.3.1, and consolidation 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-binding protein molecule (e.g., ApoA-I) as a monotherapy or as part of a combination therapy with one or more drug therapies, for example, in combination with standard treatment therapy for the disease or condition of interest. Combination therapy is described in Section 6.4.

[0148] 6.1. Lipid-binding protein molecules and lipid-binding protein-based complexes 6.1.1. Lipid-binding protein molecules Lipid-binding protein molecules that can be used directly or in the lipid-binding protein-based complexes described herein include apolipoproteins, such as those described in Section 6.1.1.1, and apolipoprotein-mimetic peptides, such as those described in Section 6.1.1.2. In some embodiments, a mixture of lipid-binding protein molecules can be used, optionally as a member of a complex. In some embodiments, the mixture of lipid-binding protein molecules can include one or more apolipoproteins. In some embodiments, the mixture of lipid-binding protein molecules can include one or more apolipoproteins. In some embodiments, the mixture of lipid-binding protein molecules can include one or more apolipoproteins and one or more apolipoprotein-mimetic peptides. In some embodiments, the mixture of lipid-binding protein molecules can include one or more apolipoproteins and one or more apolipoprotein-mimetic peptides.

[0149] Apolipoproteins The lipid-binding protein molecule can be selected, and suitable apolipoproteins that can be included in the lipid-binding protein-based complexes disclosed herein include apolipoproteins ApoA-I, ApoA-II, ApoA-IV, ApoA-V, ApoB, ApoC-I, ApoC-II, ApoC-III, ApoD, ApoE, ApoJ, ApoH, and any combination of two or more of the above. Polymorphic forms, isoforms, variants, and mutants, as well as truncated forms of the above apolipoproteins, can also be used, the most common of which is apolipoprotein AI. ミラノ (ApoA-I M ), apolipoprotein AI パリ (ApoA-I P ), and apolipoprotein AI サラゴサ (ApoA-I Z), which can be used. Also, apolipoprotein variants containing cysteine ​​residues are known and can also be used (see, for example, U.S. Patent Application Publication No. 2003 / 0181372). Apolipoproteins can be in the form of monomers or dimers, which can be homodimers or heterodimers. For example, ApoA-I (Duverger et al., 1996, Arterioscler. Thromb. Vasc. Biol. 16(12):1424-29), ApoA-I M (Franceschini et al.,1985,J.Biol.Chem.260:1632-35), ApoA-I P (Daum et al., 1999, J. Mol. Med. 77:614-22), ApoA-II (Shelness et al., 1985, J. Biol. Chem. 260(14):8637-46; Shelness et al., 1984, J. Biol. Chem. 259(15):9929-35), ApoA-IV (Duverger et al., 1991, Euro. J. Biochem. 201(2):373-83), ApoE (McLean et al., 1983, J. Biol. Chem. 258(14):8993-9000), ApoJ, and homo- and heterodimers of ApoH (where feasible) may also be used.

[0150] Apolipoproteins can be modified in their primary sequence to make them less susceptible to oxidation, as described, for example, in U.S. Patent Application 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, apolipoproteins or apolipoprotein-containing complexes are soluble in biological fluids (e.g., lymph, cerebrospinal fluid, vitreous humor, aqueous humor, blood, or blood fractions (e.g., serum or plasma)).

[0151] In some embodiments, the lipid binding protein molecule is a covalently linked lipid binding protein monomer, such as a dimeric apolipoprotein AI. ミラノ which is a mutant form of ApoA-I that contains a cysteine. The cysteine ​​allows for the formation of disulfide bridges, which can form homodimers or heterodimers (e.g., ApoA-I ミラノ This can result in the formation of ApoA-II).

[0152] 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.

[0153] 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 an ApoA-I molecule. ミラノ No.

[0154] In some embodiments, the ApoA-I molecule is apolipoprotein AI ミラノ (ApoA-I M ), apolipoprotein AI パリ (ApoA-I P ), or apolipoprotein AI サラゴサ (ApoA-I Z )

[0155] Apolipoproteins can be purified from animal sources (particularly human sources) or recombinantly produced, as is well known in the art, see, e.g., 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, and 6,617,134; U.S. Patent Application Publication Nos. 2002 / 0156007, 2004 / 0067873, 2004 / 0077541, and 2004 / 0266660; and PCT Publication Nos. 2008 / 104890 and 2007 / 023476. Other purification methods are also possible and are described, for example, in PCT Publication No. WO 2012 / 109162, the disclosure of which is incorporated herein by reference in its entirety.

[0156] In certain embodiments, the ApoA-I is recombinant ApoA-I produced by a mammalian host cell. The host cell can be from any mammalian cell line. The polynucleotide encoding ApoA-I can be codon-optimized for expression in the recombinant host cell. In some embodiments, the host cell is a mammalian host cell, including, but not limited to, Chinese hamster ovary cells (e.g., CHO-K1; ATCC No. CCL 61; CHO-S (e.g., GIBCO Life Technologies Inc., Rockville, MD, Catalog No. 11619012)), VERO cells, BHK (ATCC No. CRL 1632), BHK 570 (ATCC No. CRL 10314), HeLa cells, COS-1 (ATCC No. CRL 1650), COS-7 (ATCC No. CRL 1651), MDCK cells, 293 cells (ATCC No. CRL 1573; Graham et al., J. Gen. Virol. 36:59-72, 1977), 3T3 cells, myeloma cells (particularly murine), PC12 cells, and W138 cells. In certain embodiments, the mammalian cells, e.g., CHO-S cells, are adapted to growth in serum-free medium. Additional suitable cell lines are known in the art and available from public depositories, e.g., the American Type Culture Collection, Manassas, Va.

[0157] In some embodiments, recombinant ApoA-I is produced by CHO cells, e.g., CHO-S cells. As one skilled in the art will recognize, recombinant polypeptides (e.g., recombinant ApoA-I) expressed by mammalian host cells, e.g., CHO cells, may be subject to post-translational processing (e.g., glycosylation, etc.). The resulting recombinant ApoA-I may have one or more structural features (e.g., glycosylation pattern) that differ from ApoA-I purified from human plasma.

[0158] The apolipoprotein can be in a prepro-form, a pro-form, or a mature form. For example, the apolipoprotein can include ApoA-I (e.g., human ApoA-I), where ApoA-I is prepro-ApoA-I, pro-ApoA-I, or mature ApoA-I. In some embodiments, ApoA-I has at least 90% sequence identity with SEQ ID NO: 1 below. PPQSPWDRVKDLATVYVDVLKDSGRDYVSQFEGSALGKQLNLKLLDNWDSVTSTFSKLREQLGPVTQEFWDNLEKETEGLRQEMSKDLEEVKAKVQPYLDDFQKKWQEEMELYRQKVEPLRAELQEGARQKLHELQEKLSPLGEEMRDRARAHVDALRTHLAPYSDELRQRLAARLEALKENGGARLAEYHAKATEHLSTLSEKAKPALEDLRQGLLPVLESFKVSFLSALEEYTKKLNTQ (SEQ ID NO: 1)

[0159] In another embodiment, ApoA-I has at least 95% sequence identity to SEQ ID NO: 1. In another embodiment, ApoA-I has at least 98% sequence identity to SEQ ID NO: 1. In another embodiment, ApoA-I has at least 99% sequence identity to SEQ ID NO: 1. In another embodiment, ApoA-I has 100% sequence identity to SEQ ID NO: 1.

[0160] In some embodiments, the ApoA-I has at least 95% sequence identity to amino acids 25-267 of SEQ ID NO:2 below. MKAAVLTLAVLFLTGSQARHFWQQDEPPQSPWDRVKDLATVYVDVLKDSGRDYVSQFEGSALGKQLNLKLLDNWDSVTSTFSKLREQLGPVTQEFWDNLEKETEGLRQEMSKDLEEVKAKVQPYLDDFQKKWQEEMELYRQKVEPLRAELQEGARQKLHELQEKLSPLGEEMRDRARAHVDALRTHLAPYSDELRQRLAARLEALKENGGARLAEYHAKATEHLSTLSEKAKPALEDLRQGLLPVLESFKVSFLSALEEYTKKLNTQ (SEQ ID NO: 2)

[0161] In another embodiment, ApoA-I has at least 98% sequence identity to amino acids 25-267 of SEQ ID NO: 2. In another embodiment, ApoA-I has at least 99% sequence identity to amino acids 25-267 of SEQ ID NO: 2. In another embodiment, ApoA-I has 100% sequence identity to amino acids 25-267 of SEQ ID NO: 2. Amino acids 25-267 of SEQ ID NO: 2 are also identified herein as SEQ ID NO: 3. DEPPQSPWDRVKDLATVYVDVLKDSGRDYVSQFEGSALGKQLNLKLLDNWDSVTSTFSKLREQLGPVTQEFWDNLEKETEGLRQEMSKDLEEVKAKVQPYLDDFQKKWQEEMELYRQKVEPLRAELQEGARQKLHELQEKLSPLGEEMRDRARAHVDALRTHLAPYSDELRQRLAARLEALKENGGARLAEYHAKATEHLSTLSEKAKPALEDLRQGLLPVLESFKVSFLSALEEYTKKLNTQ (SEQ ID NO: 3).

[0162] In certain embodiments in which ApoA-I is recombinantly expressed by a host cell, the host cell comprises a nucleotide sequence encoding the amino acid sequence of a mature ApoA-I protein, which can be operably linked to a signal sequence and / or a proprotein sequence for secretion of ApoA-I from the host cell. In some embodiments, the nucleotide sequence encodes an amino acid sequence having at least 90% (e.g., at least 95%, at least 97%, or 100%) sequence identity to SEQ ID NO:2 (human preproApoA-I), which includes the signal sequence (amino acids 1-18 of SEQ ID NO:2) and the propeptide sequence (amino acids 19-24 of SEQ ID NO:2). Other signal sequences suitable for directed secretion of ApoA-I can be heterologous to ApoA-I, e.g., human albumin signal peptide or human IL-2 signal peptide, or can be homologous to ApoA-I.

[0163] In some embodiments, the nucleotide sequence encodes an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 2. In some embodiments, the nucleotide sequence encodes an amino acid sequence having at least 98% sequence identity to SEQ ID NO: 2. In some embodiments, the nucleotide sequence encodes an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 2. In some embodiments, the nucleotide sequence encodes an amino acid sequence having 100% sequence identity to SEQ ID NO: 2.

[0164] In some embodiments, ApoA-I is produced by CHO cells (e.g., CHO-S cells) engineered to express the amino acid sequence of SEQ ID NO: 2. The engineered cells can comprise a nucleotide sequence encoding SEQ ID NO: 2 operably linked to a promoter, e.g., a constitutive promoter. In some embodiments, the engineered cells can comprise a nucleotide sequence encoding SEQ ID NO: 2 operably linked to a simian cytomegalovirus immediate early promoter.

[0165] In some embodiments, recombinant ApoA-I can be produced by culturing any of the mammalian host cells described herein under conditions in which ApoA-I is expressed and secreted. ApoA-I can be recovered from the supernatant of the cultured mammalian host cells and, optionally, purified to obtain biologically active, mature ApoA-I.

[0166] Further methods for recombinant expression and purification of ApoA-I are described in detail in WO2012 / 109162, the contents of which are incorporated herein by reference in their entirety (see, e.g., Sections 6.1.2-6.1.4 and Examples 1-2 of PCT Publication No. WO2012 / 109162, the contents of which are incorporated herein by reference in their entirety).

[0167] The apolipoprotein molecule(s) can include a chimeric apolipoprotein, which comprises an apolipoprotein and one or more attached functional moieties, such as one or more CRN-001 conjugates, one or more targeting moieties, a moiety with a desired biological activity, an affinity tag to aid in purification, and / or a reporter molecule for characterization or localization studies. The attached biologically active moiety can have an activity that can enhance and / or synergize with the biological activity of a compound incorporated into the conjugate of the present disclosure. For example, the biologically active moiety can have antibacterial (e.g., antifungal, antibacterial, antiprotozoal, bacteriostatic, fungistatic, or antiviral) activity. In one embodiment, the attached functional moiety of the chimeric apolipoprotein does not contact a hydrophobic surface of the conjugate. In another embodiment, the attached functional moiety contacts a hydrophobic surface of the conjugate. In some embodiments, the functional moiety of the chimeric apolipoprotein can be native to the native protein. In some embodiments, the chimeric apolipoprotein comprises a ligand or sequence that is capable of being recognized by or interacting with a cell surface receptor or other cell surface moiety.

[0168] In one embodiment, the chimeric apolipoprotein includes a targeting moiety that is not native to the native apolipoprotein, such as S. cerevisiae alpha-mating factor peptide, folic acid, transferrin, or lactoferrin. In another embodiment, the chimeric apolipoprotein includes a moiety with a desired biological activity that enhances and / or synergizes with the activity of a compound incorporated into the complex of the present disclosure. In one embodiment, the chimeric apolipoprotein may include a functional moiety unique to the apolipoprotein. One example of an apolipoprotein-specific functional moiety is the unique targeting moiety formed by approximately amino acids 130-150 of human ApoE, which includes the receptor-binding region recognized by members of the low-density lipoprotein receptor family. Other examples of apolipoprotein-specific functional moieties include the region of ApoB-100 that interacts with the low-density lipoprotein receptor and the region of ApoA-I that interacts with scavenger receptor type B1. In other embodiments, functional moieties may be added synthetically or recombinantly to produce the chimeric apolipoprotein. Another example is an apolipoprotein having a prepro or pro sequence from another preproapolipoprotein (e.g., a prepro sequence from preproapoA-II substituting for the prepro sequence of preproapoA-I). Another example is an apolipoprotein in which some of the amphipathic sequence segments are replaced by other amphipathic sequence segments from another apolipoprotein.

[0169] As used herein, "chimera" refers to two or more molecules that can exist separately and that are linked together to form a single molecule possessing all the desired functionality of its constituent molecules. The constituent molecules of a chimeric molecule may be synthetically linked by chemical conjugation, or, if the constituent molecules are all polypeptides or their analogs, the polynucleotides encoding the polypeptides may be recombinantly fused together, resulting in the expression of a single, continuous polypeptide. Such chimeric molecules are referred to as fusion proteins. A "fusion protein" is a chimeric molecule in which the constituent molecules are all polypeptides and are attached (fused) to each other, thereby forming a continuous, single chain. The various constituents can be directly attached to each other or coupled via one or more linkers. One or more segments of the various constituents can be inserted, for example, into the apolipoprotein sequence or, as another example, added to the apolipoprotein sequence at the N- or C-terminus. For example, the fusion protein can comprise an antibody light chain, an antibody fragment, a heavy chain antibody, or a single domain antibody.

[0170] In some embodiments, chimeric apolipoproteins are prepared by chemically conjugating an apolipoprotein and a functional moiety to be attached. Means for chemically conjugating molecules are well known to those skilled in the art. Such means will vary according to the structure of the moiety to be attached, but will be readily ascertainable to those skilled in the art. Polypeptides typically contain various functional groups, such as carboxylic acid (--COOH), free amino (--NH), or sulfhydryl (--SH) groups, which are available for reaction with suitable functional groups on the functional moiety or on a linker to attach the moiety thereto. The functional moiety may be attached at the N-terminus, C-terminus, or to a functional group on an internal residue (i.e., a residue intermediate between the N- and C-terminus) of the apolipoprotein molecule. Alternatively, the apolipoprotein and / or the moiety to be tagged can be derivatized to expose or attach additional reactive functional groups.

[0171] In some embodiments, fusion proteins containing polypeptide functional portions are synthesized using a recombinant expression system. Typically, this involves creating nucleic acid (e.g., DNA) sequences encoding the 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.

[0172] The nucleic acid encoding the chimeric apolipoprotein can be incorporated into a recombinant expression vector in a form suitable for expression in a host cell. As used herein, an "expression vector" is a nucleic acid that can be transcribed and translated into a polypeptide when introduced into an appropriate host cell. The vector can also contain regulatory sequences, such as promoters, enhancers, or other expression control elements (e.g., polyadenylation signals). Such regulatory sequences are known to those skilled in the art (see, for example, Goeddel, 1990, Gene Expression Technology: Meth. Enzymol. 185, Academic Press, San Diego, Calif.; Berger and Kimmel, Guide to Molecular Cloning Techniques, Methods in Enzymology 152, Academic Press, Inc., San Diego, Calif.; Sambrook et al., 1989, Molecular Cloning—A Laboratory Manual (2nd ed.) Vol. 1-3, Cold Spring Harbor Laboratory, Cold Spring Harbor Press, NY, etc.).

[0173] In some embodiments, the apolipoprotein is modified so that, when the apolipoprotein is incorporated into a complex of the present disclosure, the modification increases the stability of the complex, confers targeting ability, or enhances its ability. In one embodiment, the modification involves introducing a cysteine ​​residue into the apolipoprotein molecule, for example, by site-directed mutagenesis, to enable intramolecular or intermolecular disulfide bond formation. In another embodiment, a chemical crosslinker is used to form an intermolecular linkage between apolipoprotein molecules to enhance the stability of the complex. The intermolecular crosslink prevents or reduces dissociation of the apolipoprotein molecule from the complex and / or prevents replacement by endogenous apolipoprotein molecules in the individual to whom the complex is administered. In other embodiments, the apolipoprotein is modified by either chemical derivatization or site-directed mutagenesis of one or more amino acid residues to confer targeting ability to or recognition by a cell surface receptor.

[0174] Lipid-binding protein molecules and complexes containing lipid-binding protein molecules can be engineered to target specific cell surface receptors by engineering receptor recognition properties into apolipoproteins. For example, lipid-binding protein molecules or complexes can be targeted to specific cell types known to harbor specific types of infectious agents by, for example, modifying the apolipoprotein of another lipid-binding protein molecule so that the apolipoprotein can interact with a receptor on the surface of the targeted cell type. For example, lipid-binding protein molecules or complexes can be targeted to macrophages by modifying the apolipoprotein or other lipid-binding protein molecule to confer recognition by macrophage endocytosis class A scavenger receptor (SR-A). SR-A binding ability can be conferred to lipid-binding protein molecules or complexes by modifying the apolipoprotein or other lipid-binding protein molecule 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 concentration of negatively charged residues. Lipid-binding protein molecules and complexes containing lipid-binding protein molecules (e.g., apolipoproteins) can also interact with apolipoprotein receptors, including, but not limited to, ABCA1 receptor, ABCG1 receptor, megalin, cubulin, and HDL receptors, such as SR-B1.

[0175] 6.1.1.2. Apolipoprotein Mimetics Peptides, peptide analogs, and agonists that mimic the activity of apolipoproteins (collectively referred to herein as "apolipoprotein peptidomimetics") can also be used as lipid-binding protein molecules or in the conjugates described herein, either alone or in combination with one or more other lipid-binding proteins. Peptides and peptide analogs corresponding to apolipoproteins, as well as ApoA-I, ApoA-I M Non-limiting examples of agonists that mimic the activity of ApoA-II, ApoA-IV, and ApoE, are useful as lipid binding protein molecules, and / or are suitable for inclusion in the conjugates and compositions described herein are described in U.S. Pat. Nos. 6,004,925, 6,037,323, and 6,046,166 (Dasseux et al., 2002). No. 5,840,688 (issued to Tso), U.S. Pat. No. 6,743,778 (issued to Kohno), U.S. Patent Application Publication Nos. 2004 / 0266671, 2004 / 0254120, 2003 / 0171277, and 2003 / 0045460 (to Fogelman), U.S. Patent Application Publication No. 2006 / 0069030 (to Bachovchin), U.S. Patent Application Publication No. 2003 / 0087819 (to Bielicki), U.S. Patent Application Publication No. 2009 / 0081293 (to Murase et al.), and PCT Publication No. WO 2010 / 093918 (to Dasseux et al.), the disclosures of which are incorporated herein by reference in their entireties. These peptides and peptide analogs can be composed of L-amino acids or D-amino acids, or a mixture of L- and D-amino acids. These peptides and peptide analogs can also contain one or more non-peptide or amide bonds, such as one or more well-known peptide / amide isosteres. Such apolipoprotein peptidomimetics can be synthesized or produced using any technique for 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.

[0176] In some embodiments, the lipid binding protein molecule comprises an apolipoprotein peptidomimetic molecule, optionally comprising one or more apolipoprotein molecules, such as those described above.

[0177] In some embodiments, the apolipoprotein peptidomimetic molecule comprises an ApoA-I peptidomimetic, an ApoA-II peptidomimetic, an ApoA-IV peptidomimetic, or an ApoE peptidomimetic, or a combination thereof.

[0178] 6.1.2. Lipid-binding protein-based conjugates In some embodiments, the lipid-binding protein is a component of a lipid-binding protein-based complex, for example, complexed with one or more amphiphilic molecules, for example, lipids (for example, formulated as a lipid-binding protein-based complex).The lipid-binding protein-based complex that can be used includes HDL and HDL mimic-based complexes.

[0179] In some embodiments, the lipid-binding protein-based complex can include the lipoprotein complexes described in U.S. Patent No. 8,206,750, PCT International Publication No. 2012 / 109162, PCT International Publication No. 2015 / 173633 A2 (e.g., CER-001), or US2004 / 0229794 A1, the contents of each of which are incorporated herein by reference in their entirety. The terms "lipoprotein" and "apolipoprotein" are used interchangeably herein, and unless otherwise required by context, the term "lipoprotein" encompasses lipoprotein mimetics. The terms "lipid-binding protein" and "lipid-binding polypeptide" are also used interchangeably herein, and unless otherwise required by context, the terms do not refer to a specific length of amino acid sequence.

[0180] The lipoprotein complex can include a protein fraction (e.g., an apolipoprotein fraction) and a lipid fraction (e.g., a phospholipid fraction). The protein fraction includes one or more lipid-binding protein molecules, e.g., an apolipoprotein, a peptide, or an apolipoprotein peptide analog or mimetic, e.g., one or more lipid-binding protein molecules described in Section 6.1.1.

[0181] The lipid fraction typically comprises one or more phospholipids, which can be neutral, negatively charged, positively charged, or a combination thereof. Exemplary phospholipids and other amphipathic molecules that can be contained within the lipid fraction are described in Section 6.1.3.

[0182] In certain embodiments, the lipid fraction contains at least one neutral phospholipid (e.g., sphingomyelin (SM)) and, optionally, one or more negatively charged phospholipids. In lipoprotein complexes containing both neutral and negatively charged phospholipids, the neutral and negatively charged phospholipids can have fatty acid chains with the same or different numbers of carbons and the same or different degrees of saturation. In some cases, the neutral and negatively charged phospholipids have the same acyl tail, e.g., C16:0, or palmitoyl acyl chain. In specific embodiments, particularly those in which egg SM is used as the neutral lipid, the weight ratio of apolipoprotein fraction:lipid fraction ranges from about 1:2.7 to about 1:3 (e.g., 1:2.7).

[0183] Any phospholipid that has at least a partial negative charge at physiological pH can be used as the negatively charged phospholipid.Non-limiting examples include phosphatidylinositol, phosphatidylserine, phosphatidylglycerol, and the negatively charged forms, such as salts, of phosphatidic acid.In a specific embodiment, the negatively charged phospholipid is 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] or DPPG, phosphatidylglycerol.Preferred salts include potassium and sodium salts.

[0184] In some embodiments, the lipoprotein complexes used in the methods of the present disclosure are those described in U.S. Patent No. 8,206,750 or WO2012 / 109162 (and its U.S. counterpart, US2012 / 0232005), the contents of each of which are incorporated herein by reference in their entirety. In certain embodiments, the lipid-binding protein molecular components of the lipoprotein complex are those described in Section 6.1, preferably Section 6.1.1, of WO2012 / 109162 (and US2012 / 0232005), and the lipid components are those described in Section 6.2 of WO2012 / 109162 (and US2012 / 0232005), which can optionally be complexed together in amounts described in Section 6.3 of WO2012 / 109162 (and US2012 / 0232005). The contents of each of these sections are incorporated herein by reference. In certain embodiments, the lipoprotein complexes of the present disclosure are in a population of complexes that is at least 85%, at least 90%, at least 95%, at least 97%, or at least 99% homogeneous, as described in Section 6.4 of WO2012 / 109162 (and US2012 / 0232005), the contents of which are incorporated herein by reference.

[0185] In some embodiments, the lipid-binding protein-based 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.

[0186] In some embodiments, the complex comprises 1 to 8 ApoA-I equivalents (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, 2 to 8, 2 to 6, 2 to 4, 4 to 6, or 4 to 8 ApoA-I equivalents). Lipid binding proteins can be expressed in terms of ApoA-I equivalents based on the number of amphipathic helices they contain. For example, ApoA-I, which typically exists as a disulfide-bridged dimer, is a soluble form of ApoA-I. M ApoA-I M Each molecule of a peptidomimetic contains twice the number of amphipathic helices as a molecule of ApoA-I, and therefore can be represented as 2 ApoA-I equivalents. Conversely, a peptidomimetic containing a single amphipathic helix can be represented as 1 / 10 to 1 / 6 ApoA-I equivalents, since each molecule contains 1 / 10 to 1 / 6 the number of amphipathic helices as a molecule of ApoA-I.

[0187] In a specific embodiment, a lipoprotein complex that can be used in the methods of the present disclosure comprises 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 50-80 lecithin molecules, and 20-50 SM molecules.

[0188] In another specific embodiment, a lipoprotein complex that can be used in the methods of the present disclosure comprises 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 50 lecithin molecules, and 50 SM molecules.

[0189] In yet another specific embodiment, a lipoprotein complex that can be used in the methods of the present disclosure comprises 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 80 lecithin molecules, and 20 SM molecules.

[0190] In yet another specific embodiment, a lipoprotein complex that can be used in the methods of the present disclosure comprises 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 70 lecithin molecules, and 30 SM molecules.

[0191] In yet another specific embodiment, a lipoprotein complex that can be used in the methods of the present disclosure comprises 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 60 lecithin molecules, and 40 SM molecules.

[0192] In a specific embodiment, a lipoprotein complex that can be used in the methods of the present disclosure consists essentially of 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 50-80 lecithin molecules, and 20-50 SM molecules.

[0193] In another specific embodiment, the lipoprotein complex that can be used in the methods of the present disclosure consists essentially of 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 50 lecithin molecules, and 50 SM molecules.

[0194] In yet another specific embodiment, a lipoprotein complex that can be used in the methods of the present disclosure consists essentially of 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 80 lecithin molecules, and 20 SM molecules.

[0195] In yet another specific embodiment, a lipoprotein complex that can be used in the methods of the present disclosure consists essentially of 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 70 lecithin molecules, and 30 SM molecules.

[0196] In yet another specific embodiment, a lipoprotein complex that can be used in the methods of the present disclosure consists essentially of 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 60 lecithin molecules, and 40 SM molecules.

[0197] In specific embodiments, lipoprotein complexes that can be used in the methods of the present disclosure include a lipid component that comprises about 90-99.8% by weight SM and about 0.2-10% by weight negatively charged phospholipids, e.g., about 0.2-1%, 0.2-2%, 0.2-3%, 0.2-4%, 0.2-5%, 0.2-6%, 0.2-7%, 0.2-8%, 0.2-9%, or 0.2-10% by weight total negatively charged phospholipid(s). In another specific embodiment, the lipoprotein complex that can be used in the methods of the present disclosure comprises about 90-99.8% by weight lecithin and about 0.2-10% by weight negatively charged phospholipid, e.g., about 0.2-1%, 0.2-2%, 0.2-3%, 0.2-4%, 0.2-5%, 0.2-6%, 0.2-7%, 0.2-8%, 0.2-9%, or 0.2-10% by weight total negatively charged phospholipid(s).

[0198] In specific embodiments, lipoprotein complexes that can be used in the methods of the present disclosure include a lipid component that consists essentially of about 90-99.8% by weight SM and about 0.2-10% by weight negatively charged phospholipids, e.g., about 0.2-1%, 0.2-2%, 0.2-3%, 0.2-4%, 0.2-5%, 0.2-6%, 0.2-7%, 0.2-8%, 0.2-9%, or 0.2-10% by weight total negatively charged phospholipid(s). In another specific embodiment, the lipoprotein complexes that can be used in the methods of the present disclosure consist essentially of about 90-99.8% by weight lecithin and about 0.2-10% by weight negatively charged phospholipids, e.g., about 0.2-1%, 0.2-2%, 0.2-3%, 0.2-4%, 0.2-5%, 0.2-6%, 0.2-7%, 0.2-8%, 0.2-9%, or 0.2-10% by weight total negatively charged phospholipid(s).

[0199] In yet another specific embodiment, the lipoprotein complexes that can be used in the methods of the present disclosure comprise a lipid fraction that contains about 9.8-90% by weight SM, about 9.8-90% by weight lecithin, and about 0.2-10% by weight negatively charged phospholipids, e.g., from about 0.2-1% by weight, to 0.2-2%, 0.2-3%, 0.2-4%, 0.2-5%, 0.2-6%, 0.2-7%, 0.2-8%, 0.2-9%, or 0.2-10% by weight total negatively charged phospholipid(s).

[0200] In yet another specific embodiment, the lipoprotein complexes that can be used in the methods of the present disclosure comprise a lipid fraction that consists essentially of about 9.8-90% by weight SM, about 9.8-90% by weight lecithin, and about 0.2-10% by weight negatively charged phospholipids, e.g., from about 0.2-1% by weight, to 0.2-2%, 0.2-3%, 0.2-4%, 0.2-5%, 0.2-6%, 0.2-7%, 0.2-8%, 0.2-9%, or 0.2-10% by weight total negatively charged phospholipid(s).

[0201] In another specific embodiment, lipoprotein complexes that can be used in the methods of the present disclosure comprise ApoA-I apolipoprotein and a lipid fraction, the lipid fraction comprising sphingomyelin and about 3% by weight of negatively charged phospholipids, the molar ratio of lipid fraction to ApoA-I apolipoprotein is about 2:1 to 200:1, and the complexes are small or large discoid particles containing 2 to 4 ApoA-I equivalents.

[0202] In another specific embodiment, the lipoprotein complexes that can be used in the methods of the present disclosure comprise ApoA-I apolipoprotein and a lipid fraction, wherein the lipid fraction consists essentially of sphingomyelin and about 3% by weight of negatively charged phospholipids, the molar ratio of lipid fraction to ApoA-I apolipoprotein is about 2:1 to 200:1, and the complexes are small or large discoid particles containing 2 to 4 ApoA-I equivalents.

[0203] The HDL-based or HDL mimetic-based complex can comprise a single type of lipid-binding protein, or a mixture of two or more different lipid-binding proteins, which can be derived from the same or different species.Although not required, the complex preferably comprises a lipid-binding protein derived from the animal species to be treated or corresponding to it in amino acid sequence, in order to avoid inducing an immune response to therapy.Therefore, 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 immune response.

[0204] In some embodiments, the lipid component comprises two types of phospholipids: sphingomyelin (SM) and negatively charged phospholipids. Exemplary SMs and negatively charged lipids are described in Section 6.1.3.1.

[0205] The lipid component comprising SM can optionally include small amounts of additional lipids. Virtually any type of lipid may be used, including, but not limited to, lysophospholipids, galactocerebrosides, gangliosides, cerebrosides, glycerides, triglycerides, and cholesterol, and derivatives thereof.

[0206] When included, such optional lipids typically comprise less than about 15% by weight of the lipid fraction, although in some cases, more optional lipids may be included. In some embodiments, optional lipids comprise less than about 10% by weight, less than about 5% by weight, or less than about 2% by weight. In some embodiments, the lipid fraction does not comprise optional lipids.

[0207] In a specific embodiment, the phospholipid fraction contains egg SM or palmitoyl SM or phytosphingomyelin and DPPG in a weight ratio (SM:negatively charged phospholipids) ranging from 90:10 to 99:1, more preferably ranging from 95:5 to 98:2. In one embodiment, the weight ratio is 97:3.

[0208] The molar ratio of lipid to protein components of the complexes of the present disclosure can vary and depends, among other factors, on the identity(ies) of the apolipoprotein(s) comprising the protein component, the identity and amount of the lipid(s) comprising the lipid component, and the desired size of the complex. Because the biological activity of apolipoproteins, such as ApoA-I, is believed to be mediated by the amphipathic helices that comprise the apolipoprotein, it is convenient to express the apolipoprotein fraction of the lipid:apolipoprotein molar ratio using ApoA-I protein equivalents. ApoA-I is generally accepted to contain 6 to 10 amphipathic helices, depending on the method used to calculate helices. Other apolipoproteins can be expressed in terms of ApoA-I equivalents, based on the number of amphipathic helices they contain. For example, ApoA-I, which typically exists as a disulfide-bridged dimer, is expressed in terms of ApoA-I equivalents, based on the number of amphipathic helices it contains.M ApoA-I M Each molecule of a lipoprotein contains twice the number of amphipathic helices as a molecule of ApoA-I, and therefore can be expressed as 2 ApoA-I equivalents. Conversely, a peptide apolipoprotein containing a single amphipathic helix can be expressed as 1 / 10 to 1 / 6 ApoA-I equivalents, since each molecule contains 1 / 10 to 1 / 6 of the amphipathic helices as a molecule of ApoA-I. Generally, the lipid:ApoA-I equivalent molar ratio (defined herein as "Ri") of lipoprotein complexes ranges from about 105:1 to 110:1. In some embodiments, Ri is about 108:1. Ratios by weight can be obtained using MWs of approximately 650 to 800 for phospholipids.

[0209] In some embodiments, the lipid:ApoA-I equivalent molar ratio ("RSM") ranges from about 80:1 to about 110:1, e.g., from about 80:1 to about 100:1. In a specific example, the RSM for the complex can be about 82:1.

[0210] In some embodiments, the lipoprotein complex used in the methods of the present disclosure is a negatively charged complex, which comprises a protein fraction, preferably a protein fraction that is mature full-length ApoA-I, and a lipid fraction comprising neutral phospholipids, sphingomyelin (SM), and negatively charged phospholipids.

[0211] In a specific embodiment, the lipid component contains SM (e.g., egg SM, palmitoyl SM, phyto SM, or a combination thereof) and negatively charged phospholipid (e.g., DPPG) in a weight ratio (SM:negatively charged phospholipid) ranging from 90:10 to 99:1, more preferably ranging from 95:5 to 98:2, e.g., 97:3.

[0212] In specific embodiments, the ratio of protein to lipid components can range from about 1:2.7 to about 1:3, with 1:2.7 being preferred. This corresponds to 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.

[0213] In certain embodiments, the conjugate comprises CER-001, CSL-111, CSL-112, CER-522, ETC-216, or ETC-642. In a preferred embodiment, the conjugate is CER-001.

[0214] As used in the literature and in the following examples, CER-001 refers to the complex described in Example 4 of WO2012 / 109162. WO2012 / 109162 refers to CER-001 as a complex having a lipoprotein weight:total phospholipid weight ratio of 1:2.7 and an SM:DPPG weight:weight ratio of 97:3. Example 4 of WO2012 / 109162 also describes a method for its preparation.

[0215] As used in the context of the methods and / or CER-001 dosing regimens of the present disclosure, CER-001 refers to a lipoprotein complex whose individual constituents can vary by up to 20% from CER-001 described in Example 4 of WO2012 / 109162. In certain embodiments, the constituents of the lipoprotein complex vary by up to 10% from CER-001 described in Example 4 of WO2012 / 109162. Preferably, the constituents of the lipoprotein complex are those described in Example 4 of WO2012 / 109162 (plus / minus allowed manufacturing tolerance variations). The SM in CER-001 can be natural or synthetic. In some embodiments, the SM is a natural SM, e.g., a natural SM described in WO2012 / 109162, e.g., chicken egg SM or plant SM. In some embodiments, the SM is a synthetic SM, such as those described in WO2012 / 109162, e.g., synthetic palmitoylsphingomyelin, e.g., those described in WO2012 / 109162. Methods for synthesizing palmitoylsphingomyelin are known in the art, e.g., those described in WO2014 / 140787 and WO2024 / 003612, the contents of which are incorporated herein by reference in their entireties. The lipoprotein in CER-001, apolipoprotein AI (ApoA-I), preferably has an amino acid sequence corresponding to amino acids 25-267 of SEQ ID NO: 1 in WO2012 / 109162. ApoA-I can be purified from animal sources (particularly from human sources) 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, e.g., 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, e.g., Section 6.4 of WO2012 / 109162.

[0216] SEQ ID NO: 1 of WO2012 / 109162 is identified herein as SEQ ID NO: 2. Amino acids 25 to 267 of SEQ ID NO: 1 of WO2012 / 109162 are identified herein as SEQ ID NO: 3.

[0217] CSL-111 is a reconstituted human ApoA-I purified from plasma and complexed with soybean phosphatidylcholine (SBPC) (Tardif et al., 2007, JAMA 297:1675-1682).

[0218] CSL-112 is a preparation of ApoA-I that is purified from plasma and reconstituted to form HDL suitable for intravenous infusion (Diditchenko et al., 2013, DOI 10.1161 / ATVBAHA.113.301981).

[0219] ETC-216 (also known as MDCO-216) is a recombinant ApoA-I ミラノ It is a lipid-depleted form of HDL that contains (Nicholls et al., 2011, Expert Opin Biol Ther. 11(3):387-94. doi:10.1517 / 14712598.2011.557061).

[0220] ETC-642 is a complex of a 22-amino acid amphipathic peptide (ESP-2418) complexed with sphingomyelin and 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (dipalmitoylphosphatidylcholine, DPPC) (Di Bartolo et al., 2011, Atherosclerosis 217:395-400).

[0221] In another embodiment, the complex that can be used in the methods of the present disclosure is CER-522, a lipoprotein complex that includes a combination of three phospholipids and the following 22-amino acid peptide, CT80522: [ka] CT80522

[0222] The phospholipid components of CER-522 consist of egg sphingomyelin, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (dipalmitoylphosphatidylcholine, DPPC), and 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(l-glycerol)] (dipalmitoylphosphatidylglycerol, DPPG) in a weight ratio of 48.5:48.5:3. The peptide to total phospholipid ratio in the CER-522 complex is 1:2.5 (w / w).

[0223] In some embodiments, the lipoprotein complex is delipidated HDL. Most HDL in plasma is cholesterol-rich. Lipids in HDL can be depleted, for example, to reduce its cholesterol content, for example, partially and / or selectively depleted. In some embodiments, delipidated HDL can resemble small α, pre-β-1, and other pre-β forms of HDL. The process for selective depletion of HDL is described in Sacks et al., 2009, J Lipid Res. 50(5):894-907.

[0224] In certain embodiments, the lipoprotein complexes comprise bioactive agent delivery particles as described in US 2004 / 0229794.

[0225] The bioactive agent delivery particle can include a lipid-binding polypeptide (e.g., an apolipoprotein as previously described in this section or in Section 6.1.1), a lipid bilayer (e.g., comprising one or more phospholipids as previously described in this section or in Section 6.1.3.1), and a bioactive agent (e.g., an anticancer drug), wherein the interior of the lipid bilayer includes a hydrophobic region, and the bioactive agent is associated with the hydrophobic region of the lipid bilayer. In some embodiments, the bioactive agent delivery particle is described in US 2004 / 0229794.

[0226] In some embodiments, the bioactive agent delivery particles do not include a hydrophilic core.

[0227] In some embodiments, the bioactive agent delivery particles are disc-shaped (eg, having a diameter of about 7 to about 29 nm).

[0228] The bioactive agent delivery particle comprises a bilayer-forming lipid, e.g., a phospholipid (e.g., those described previously in this section or in Section 6.1.3.1). In some embodiments, the bioactive agent delivery particle comprises both bilayer-forming and non-bilayer-forming lipids. In some embodiments, the lipid bilayer of the bioactive agent delivery particle comprises a phospholipid. In one embodiment, the phospholipid incorporated within the delivery particle comprises dimyristoyl phosphatidylcholine (DMPC) and dimyristoyl phosphatidylglycerol (DMPG). In one embodiment, the lipid bilayer comprises DMPC and DMPG in a molar ratio of 7:3.

[0229] In some embodiments, the lipid-binding polypeptide is an apolipoprotein (e.g., as described previously in this section or in Section 6.1.1). The primary interaction between a lipid-binding polypeptide, e.g., an apolipoprotein molecule, and a lipid bilayer is generally a hydrophobic interaction between residues on the hydrophobic face of the amphiphilic structure, e.g., the α-helix of the lipid-binding polypeptide, and the fatty acyl chains of lipids on the outer surface around the particle. The bioactive agent delivery particle may contain exchangeable and / or non-exchangeable apolipoproteins. In one embodiment, the lipid-binding polypeptide is ApoA-I.

[0230] In some embodiments, the bioactive agent delivery particles comprise lipid-binding polypeptide molecules, e.g., apolipoprotein molecules, that have been modified to increase the stability of the particles. In one embodiment, the modification comprises the introduction of cysteine ​​residues to form intramolecular and / or intermolecular disulfide bonds.

[0231] In another embodiment, the bioactive agent delivery particle comprises a chimeric lipid-binding polypeptide molecule, e.g., a chimeric apolipoprotein molecule, which has one or more attached functional moieties, e.g., one or more targeting moieties, and / or one or more moieties with a desired biological activity, e.g., antimicrobial activity, which may enhance or act in synergy with the activity of the bioactive agent incorporated within the delivery particle.

[0232] 6.1.3. Amphiphilic molecules Amphipathic molecules are molecules that have both hydrophobic (non-polar) and hydrophilic (polar) elements. Amphipathic molecules that can be used in the complexes described herein include lipids (e.g., as described in Section 6.1.3.1), surfactants (e.g., as described in Section 6.1.3.2), fatty acids (e.g., as described in Section 6.1.3.3), and non-polar molecules and sterols covalently attached to polar molecules, where the polar molecules are, for example, but not limited to, sugars or nucleic acids (e.g., as described in Section 6.1.3.4).

[0233] The complex can include a single class of amphipathic molecule (e.g., a single species of phospholipid or a mixture of phospholipids) or can contain a combination of multiple classes of amphipathic molecules (e.g., phospholipids and surfactants). The complex can contain one species of amphipathic molecule or a combination of amphipathic molecules configured to facilitate solubilization of the lipid-binding protein molecule(s).

[0234] In some embodiments, the included amphiphilic molecules include phospholipids, surfactants, fatty acids, apolar moieties covalently attached to sugars or sterols, or combinations thereof (e.g., selected from the types of amphiphilic molecules discussed above).

[0235] In some embodiments, the amphipathic molecules comprise or consist of phospholipid molecules. In some embodiments, the phospholipid molecules comprise negatively charged phospholipids, neutral phospholipids, positively charged phospholipids, or a combination thereof. In some embodiments, the phospholipid molecules contribute 1 to 3 net charges per apolipoprotein molecule in the complex. In some embodiments, the net charge is a negative net charge. In some embodiments, the net charge is a positive net charge. In some embodiments, the phospholipid molecules consist of a combination of negatively charged and neutral phospholipids. In some embodiments, the molar ratio of negatively charged phospholipids to neutral phospholipids ranges from 1:1 to 1:3. In some embodiments, the molar ratio of negatively charged phospholipids to neutral phospholipids is about 1:1 or about 1:2.

[0236] In some embodiments, the amphipathic molecule comprises a neutral phospholipid and a negatively charged phospholipid in a weight ratio of 95:5 to 99:1.

[0237] Lipids A lipid-binding protein-based complex can include one or more lipids. In various embodiments, the one or more lipids can be saturated and / or unsaturated, natural and / or synthetic, charged or uncharged, zwitterionic or non-zwitterionic. In some embodiments, the lipid molecules (e.g., phospholipid molecules) together can contribute 1 to 3 (e.g., 1 to 3, 1 to 2, 2 to 3, 1, 2, or 3) net charges per lipid-binding protein molecule in the complex. In some embodiments, the net charge is negative. In other embodiments, the net charge is positive.

[0238] In some embodiments, the lipid comprises a phospholipid. The phospholipid can have two identical or different acyl chains (for example, chains with different numbers of carbon atoms, different degrees of saturation between acyl chains, different branching of acyl chains, or a combination thereof). The lipid can also be modified to contain a fluorescent probe (for example, those listed at avantilipids.com / product-category / products / fluorescent-lipids / ). Preferably, the lipid comprises at least one phospholipid.

[0239] Phospholipids can have unsaturated or saturated acyl chains ranging from about 6 to about 24 carbon atoms (e.g., 6-20, 6-16, 6-12, 12-24, 12-20, 12-16, 16-24, 16-20, or 20-24). In some embodiments, phospholipids used in the conjugates of the present disclosure have one or two acyl chains of 12, 14, 16, 18, 20, 22, or 24 carbons (e.g., two acyl chains of the same length or two acyl chains of different lengths).

[0240] Non-limiting examples of acyl chains present in commonly occurring fatty acids that can be included in phospholipids are provided in Table 1 below. [Table 1]

[0241] Lipids that can be present in the complexes of the present disclosure include small alkyl chain phospholipids, egg phosphatidylcholine, soy phosphatidylcholine, dipalmitoylphosphatidylcholine, dimyristoylphosphatidylcholine, distearoylphosphatidylcholine, 1-myristoyl-2-palmitoylphosphatidylcholine, 1-palmitoyl-2-myristoylphosphatidylcholine, 1-palmitoyl-2-stearoylphosphatidylcholine, 1-stearoyl-2-palmito ... Dilauroylphosphatidylcholine, dioleoylphosphatidylcholine, dioleoylphosphatidylethanolamine, dilauroylphosphatidylglycerol, phosphatidylcholine, phosphatidylserine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylglycerol, diphosphatidylglycerol, e.g., dimyristoylphosphatidylglycerol, dipalmitoylphosphatidylglycerol, distearoylphosphatidylglycerol Cerol, dioleoylphosphatidylglycerol, dimyristoylphosphatidic acid, dipalmitoylphosphatidic acid, dimyristoylphosphatidylethanolamine, dipalmitoylphosphatidylethanolamine, dimyristoylphosphatidylserine, dipalmitoylphosphatidylserine, brain phosphatidylserine, brain sphingomyelin, palmitoylsphingomyelin, dipalmitoylsphingomyelin, egg sphingomyelin, milk sphingomyelin Lipids that can be used to minimize lipid oxidation include, but are not limited to, phosphate, phytosphingomyelin, distearoylsphingomyelin, dipalmitoylphosphatidylglycerol salts, phosphatidic acid, galactocerebrosides, gangliosides, cerebrosides, dilaurylphosphatidylcholine, (1,3)-D-mannosyl-(1,3) diglyceride, aminophenylglycosides, 3-cholesteryl-6'-(glycosylthio)hexyl ether glycolipids, and cholesterol, and derivatives thereof. Synthetic lipids, such as synthetic palmitoylsphingomyelin or N-palmitoyl-4-hydroxysphinganine-1-phosphocholine (a form of phytosphingomyelin), can be used to minimize lipid oxidation.

[0242] In some embodiments, the lipid-binding protein-based complex comprises two types of phospholipids: a neutral lipid, e.g., lecithin and / or sphingomyelin (abbreviated as SM), and a charged phospholipid (e.g., a negatively charged phospholipid). A "neutral" phospholipid has a net charge of about 0 at physiological pH. In many embodiments, the neutral phospholipid is zwitterionic, although other types of net-neutral phospholipids are known and can be used. In some embodiments, the molar ratio of charged phospholipid (e.g., a negatively charged phospholipid) to neutral phospholipid is in the range of 1:1 to 1:3, e.g., about 1:1, about 1:2, or about 1:3.

[0243] The neutral phospholipids can include, for example, one or both of lecithin and / or SM, and can optionally include other neutral phospholipids. In some embodiments, the neutral phospholipids include lecithin but 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. Although all of these specific exemplary embodiments can include neutral phospholipids in addition to lecithin and / or SM, many embodiments do not include such additional neutral phospholipids.

[0244] As used herein, the term "SM" includes sphingomyelin derived from or obtained from natural sources, as well as analogs and derivatives of naturally occurring SM that are not susceptible to hydrolysis by LCAT like naturally occurring SM. SM is a phospholipid that is very similar in structure to lecithin, but unlike lecithin, SM does not have a glycerol backbone and therefore does not have ester bonds to attach acyl chains. Rather, SM has a ceramide backbone with amide bonds connecting the acyl chains. SM can be obtained, for example, from milk, eggs, or brain. SM analogs or derivatives can also be used. Non-limiting examples of useful SM analogs and derivatives include, but are not limited to, palmitoylsphingomyelin, N-palmitoyl-4-hydroxysphinganine-1-phosphocholine (a form of phytosphingomyelin), palmitoylsphingomyelin, stearoylsphingomyelin, D-erythro-N-16:0-sphingomyelin, and its dihydro isomer, D-erythro-N-16:0-dihydro-sphingomyelin. Synthetic SM, such as synthetic palmitoylsphingomyelin or N-palmitoyl-4-hydroxysphinganine-1-phosphocholine (phytosphingomyelin), can be used to produce complexes that are more homogeneous and have fewer contaminants and / or oxidation products than sphingolipids of animal origin. Methods for synthesizing SM are described in U.S. Patent Application Publication No. 2016 / 0075634.

[0245] Sphingomyelin isolated from natural sources can be artificially enriched in one particular saturated or unsaturated acyl chain. For example, milk sphingomyelin (Avanti Phospholipids, Alabaster, Ala.) is characterized by long saturated acyl chains (i.e., acyl chains with 20 or more carbon atoms). In contrast, egg sphingomyelin is characterized by short saturated acyl chains (i.e., acyl chains with fewer than 20 carbon atoms). For example, only about 20% of milk sphingomyelin contains C16:0 (16-carbon, saturated) acyl chains, while about 80% of egg sphingomyelin contains C16:0 acyl chains. Using solvent extraction, the composition of milk sphingomyelin can be enriched to have an acyl chain composition equivalent to that of egg sphingomyelin, or vice versa.

[0246] SM can be semi-synthetic, whereby the SM has a specific acyl chain. For example, milk sphingomyelin can be first purified from milk, and then one specific acyl chain, e.g., the C16:0 acyl chain, can be cleaved and replaced with another acyl chain. SM can also be synthesized entirely, for example, by large-scale synthesis. See, for example, Dong et al., U.S. Patent No. 5,220,043, entitled "Synthesis of D-erythro-sphingomyelins," issued June 15, 1993; Weis, 1999, Chem. Phys. Lipids 102(1-2):3-12. SM can be fully synthesized, for example, as described in U.S. Patent Application Publication No. 2014 / 0275590.

[0247] The length and level of saturation of the acyl chains comprising semi-synthetic or synthetic SM can be selectively varied. The acyl chains can be saturated or unsaturated and can contain from about 6 to about 24 carbon atoms. Each chain can contain the same number of carbon atoms, or alternatively, each chain can contain a different number of carbon atoms. In some embodiments, semi-synthetic or synthetic SM comprises mixed acyl chains, whereby one chain is saturated and one chain is unsaturated. In such mixed acyl chain SM, the chain lengths can be the same or different. In other embodiments, the acyl chains of the semi-synthetic or synthetic SM are either both saturated or both unsaturated. Again, the chains can contain the same or different numbers of carbon atoms. In some embodiments, both acyl chains comprising the semi-synthetic or synthetic SM are identical. In specific embodiments, the chains correspond to the acyl chains of naturally occurring fatty acids, such as oleic, palmitic, or stearic acid. In another embodiment, SM with saturated or unsaturated functionalized chains is used. In another specific embodiment, both acyl chains are saturated and contain 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.

[0248] In some embodiments, the SM is palmitoyl SM having a C16:0 acyl chain, eg, synthetic palmitoyl SM, or egg SM containing palmitoyl SM as the major component.

[0249] In a specific embodiment, a functionalized SM, such as phytosphingomyelin, is used.

[0250] Lecithin can be derived from natural sources or can be isolated from them, or lecithin can be obtained synthetically.The example of suitable lecithin isolated from natural sources includes, but is not limited to, egg phosphatidylcholine and soybean phosphatidylcholine.Other non-limiting examples of suitable lecithin include dipalmitoyl phosphatidylcholine, dimyristoyl phosphatidylcholine, distearoyl phosphatidylcholine, 1-myristoyl 1-2-palmitoyl phosphatidylcholine, 1-palmitoyl 1-2-myristoyl phosphatidylcholine, 1-palmitoyl 1-2-stearoyl phosphatidylcholine, 1-stearoyl 1-2-palmitoyl phosphatidylcholine, 1-palmitoyl 1-2-oleoyl phosphatidylcholine, 1-oleoyl 1-2-palmityl phosphatidylcholine, dioleoyl phosphatidylcholine and their ether derivatives or analogues.

[0251] Lecithin derived from or isolated from natural sources can be enriched to contain designated acyl chains. In embodiments using semi-synthetic or synthetic lecithin, the identity(ies) of the acyl chains can be selectively varied, as discussed above in connection with SM. In some embodiments of the complexes described herein, both acyl chains in the lecithin are identical. In some embodiments of complexes containing both SM and lecithin, the acyl chains of the SM and lecithin are all identical. In specific embodiments, the acyl chains correspond to the acyl chains of myristicin, palmitic, oleic, or stearic acid.

[0252] The complexes of the present disclosure can include one or more negatively charged phospholipids (e.g., alone or in combination with one or more neutral phospholipids). As used herein, a "negatively charged phospholipid" is a phospholipid that has a net negative charge at physiological pH. The negatively charged phospholipid can include a single type of negatively charged phospholipid or a mixture of two or more 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, phospatidyl inositol, phosphatidylserine, phosphatidic acid, and salts thereof (e.g., sodium or potassium salts). In some embodiments, the negatively charged phospholipid includes one or more of phosphatidylinositol, phosphatidylserine, phosphatidylglycerol, and / or phosphatidic acid. In a specific embodiment, the negatively charged phospholipid comprises or consists of a salt of phosphatidylglycerol or a salt of phosphatidylinositol. In another specific embodiment, the negatively charged phospholipid comprises or consists of 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] or DPPG, or a salt thereof.

[0253] Negatively charged phospholipids can be obtained from natural sources or prepared by chemical synthesis. In embodiments using synthetic negatively charged phospholipids, the identity of the acyl chain can be selectively varied, as discussed above in connection with SM. In some embodiments of the complexes of the present disclosure, both acyl chains in the negatively charged phospholipids are the same. In some embodiments, all acyl chains of all types of phospholipids contained within the complexes of the present disclosure are the same. In specific embodiments, the complexes include negatively charged phospholipid(s) and / or SM, and the negatively charged phospholipid(s) and / or SM all have C16:0 or C16:1 acyl chains. In a specific embodiment, the fatty acid moiety of the SM is predominantly C16:1 palmitoyl. In one specific embodiment, the acyl chains of the charged phospholipid(s), lecithin, and / or SM correspond to the acyl chains of palmitic acid. In yet another specific embodiment, the acyl chains of the charged phospholipid(s), lecithin, and / or SM correspond to the acyl chains of oleic acid.

[0254] Examples of positively charged phospholipids that can be included in the complexes of the present disclosure include N1-[2-((1S)-1-[(3-aminopropyl)amino]-4-[di(3-amino-propyl)amino]butylcarboxamido)ethyl]-3,4-di[oleyloxy]-benzamide, 1,2-di-O-octadecenyl-3-trimethylammonium propane, 1,2-dimyristoleoyl-sn-glycero-3-ethylphosphocholine, 1-palmitoyl-2-oleoyl-sn-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-dimethyl Ammonium-propane 1,2-dimyristoyl-3-dimethylammonium-propane, 1,2-dipalmitoyl-3-dimethylammonium-propane, N-(4-carboxybenzyl)-N,N-dimethyl-2,3-bis(oleoyloxy)propan-1-aminium, 1,2-dioleoyl-3-trimethylammonium-propane, 1,2-dioleoyl-3-trimethylammonium-propane, 1,2-stearoyl-3-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)-1propanaminium methylsulfate, and salts thereof (e.g., chloride or bromide salts).

[0255] The lipids used are preferably at least 95% pure and / or have reduced levels of oxidizing agents (e.g., but not limited to, peroxides). Lipids obtained from natural sources preferably have fewer polyunsaturated fatty acid moieties and / or fatty acid moieties that are less susceptible to oxidation. The level of oxidation in a sample can be determined using an iodometric titration method, which provides a peroxide value, expressed as milliequivalents of isolated iodine per kg of sample, abbreviated as meq O / kg. See, for example, Gray, 1978, Journal of the American Oil Chemists Society 55:539-545; Heaton et al., 1958, Journal of the Science of Food and Agriculture 9:781-786. Preferably, the level of oxidation or peroxide 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.

[0256] In some embodiments, the complex may contain a small amount of additional lipid. Substantially 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 complex of the present disclosure may contain cholesterol or a cholesterol derivative, such as a cholesterol ester. The cholesterol derivative may also be a substituted cholesterol or a substituted cholesterol ester. The complex of the present disclosure may also contain an oxidized sterol, such as, but not limited to, oxidized cholesterol or an oxidized sterol derivative (e.g., an oxidized cholesterol ester). In some embodiments, the complex does not contain cholesterol and / or its derivatives (e.g., a cholesterol ester or an oxidized cholesterol ester).

[0257] Surfactants The complex may contain one or more surfactants. The surfactant may be zwitterionic, nonionic, cationic, anionic, or a combination thereof. Exemplary zwitterionic surfactants 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 surfactants 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, and 1-decanoyl-rac-glycerol. Exemplary cationic surfactants include (S)-O-methyl-serine dodecylamide hydrochloride, dodecylammonium chloride, decyltrimethylammonium bromide, and cetyltrimethylammonium sulfate. Exemplary anionic surfactants include cholesteryl hemisuccinate, cholate, alkyl sulfates, and alkyl sulfonates.

[0258] 6.1.3.3.Fatty acids The complex can contain one or more fatty acids, including short-chain fatty acids having an aliphatic tail of 5 carbons or less (e.g., butyric acid, isobutyric acid, valeric acid, or isovaleric acid), medium-chain fatty acids having an aliphatic tail of 6 to 12 carbons (e.g., caproic acid, caprylic acid, capric acid, or lauric acid), long-chain fatty acids having an aliphatic tail of 13 to 21 carbons (e.g., myristic acid, palmitic acid, stearic acid, or arachidic acid), very long-chain fatty acids having an aliphatic tail of 22 or more carbons (e.g., behenic acid, lignoceric acid, or cerotic acid), or combinations thereof. The one or more fatty acids 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, linoelaidic 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 conjugates of the present disclosure is a cis fatty acid.

[0259] 6.1.3.4. Nonpolar molecules and sterols attached to sugars The complex can contain one or more amphiphilic molecules, which include a nonpolar molecule or moiety (e.g., a hydrocarbon chain, an 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 or substituted sugar. Exemplary amphiphilic molecules comprising a nonpolar molecule attached to a sugar include dodecan-2-yloxy-β-D-maltoside, tridecan-3-yloxy-β-D-maltoside, tridecan-2-yloxy-β-D-maltoside, n-dodecyl-β-D-maltoside (DDM), n-octyl-β-D-glucoside, n-nonyl-β-D-glucoside, n-decyl-β-D-maltoside, n-dodecyl-β-D-maltopyranoside, 4-n-dodecyl-α,α-trehalose, 6-n-dodecyl-α,α-trehalose, and 3-n-dodecyl-α,α-trehalose.

[0260] In some embodiments, the non-polar moiety is an acyl or diacyl chain.

[0261] In some embodiments, the sugar is a modified or substituted sugar.

[0262] 6.1.4. Formulations Lipid-binding protein molecules and lipid-binding protein-based complexes containing lipid-binding protein molecules 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).

[0263] CER-001 intended for administration by injection can be formulated in a phosphate buffer with sucrose and mannitol excipients, for example, as described in WO2012 / 109162.

[0264] 6.2. Target population The subjects that can be treated according to the methods described herein are preferably mammals, most preferably humans.

[0265] In some embodiments, the subject has or is at risk of having a condition associated with abnormal levels of TREM-1.Such conditions include but are not limited to acute myocardial infarction (AMI), Alzheimer's disease, chronic inflammatory bowel disease (IBD), cardiovascular disease (CVD), stroke, transient ischemic attack, transplanted organ rejection (e.g., transplanted heart rejection), ischemia-reperfusion-induced tissue injury, postoperative inflammation, psoriasis, sepsis (e.g., septic shock), and sepsis-induced acute kidney injury (AKI).

[0266] In some embodiments, the subject has or is at risk for a condition associated with abnormal levels of albumin, including, but not limited to, hypoalbuminemia (e.g., caused by liver disease, heart failure, malnutrition or vitamin deficiency, inflammatory bowel disease, kidney disease, infection, stress, thyroid disease, diabetes, nephrotic syndrome, lupus, or cirrhosis).

[0267] In some embodiments, the subject has or is at risk of having a condition associated with abnormal levels of kynurenine pathway biomarkers.Such condition includes but is not limited to Alzheimer's disease, attention deficit / hyperactivity disorder (ADHD), CNS disease, COVID-19 cognitive dysfunction, depression and major depressive disorder, epilepsy, HIV-associated neurocognitive disorder, Huntington's disease, long-term cognitive dysfunction after sepsis, death and neurological outcome after cardiac arrest, multiple sclerosis (MS), Parkinson's disease, schizophrenia, sepsis and sepsis-induced AKI.

[0268] In some embodiments, the subject has or is at risk of a condition associated with abnormal levels of IL-10.Such conditions include but are not limited to autoimmune disease, IBD, rheumatoid arthritis (RA), systemic lupus erythematosus, type I diabetes, MS, pemphigus vulgaris, ulcerative colitis (UC), Sjogren's syndrome, Graves' disease, myasthenia gravis, psoriasis, autoimmune lymphoproliferative syndrome (ALPS), cytokine release syndrome (CRS, cytokine storm), sepsis, and sepsis-induced AKI.

[0269] In some embodiments, the subject has or is at risk for a condition associated with abnormal levels of TNFα, including, but not limited to, CRS, sepsis, and sepsis-induced AKI.

[0270] In some embodiments, the subject has or is at risk for a condition associated with abnormal levels of MCP-1, including, but not limited to, CRS, sepsis, and sepsis-induced AKI.

[0271] In some embodiments, the subject has or is at risk for a condition associated with abnormal levels of IL-6, including, but not limited to, CRS, sepsis, and sepsis-induced AKI.

[0272] In some embodiments, the subject has or is at risk for a condition associated with abnormal levels of IL-8, including, but not limited to, CRS, sepsis, and sepsis-induced AKI.

[0273] In some embodiments, the subject has or is at risk for a condition associated with abnormal levels of VCAM-1 and / or ICAM-1, including, but not limited to, vascular endothelial disorders.

[0274] In some embodiments, the subject has or is at risk of bacterial infection.Examples of bacteria that commonly cause infection and sepsis include Staphylococcus aureus, Escherichia coli, Streptococcus pneumoniae, Klebsiella pneumoniae and Pseudomonas aeruginosa (GBD 2019 Antimicrobial Resistance Collaborators, 2023, Lancet 400(10369):2221-2248).

[0275] Other examples of bacteria that can cause infection and sepsis include Acinetobacter baumanni, Bacteroides fragilis, and Proteus mirabilis.

[0276] In some embodiments, the subject has or is at risk for a gram-positive bacterial infection.

[0277] In some embodiments, the subject has or is at risk for a gram-negative bacterial infection.

[0278] In some embodiments, the subject has or is at risk for a viral infection, e.g., a SARS-CoV-2 (COVID-19) infection or an influenza virus infection.

[0279] In some embodiments, the subject has or is at risk for acute myocardial infarction (AMI).

[0280] In some embodiments, the subject has or is at risk for Alzheimer's disease.

[0281] In some embodiments, the subject has or is at risk for chronic inflammatory bowel disease (IBD).

[0282] In some embodiments, the subject has or is at risk for cardiovascular disease (CVD).

[0283] In some embodiments, the subject has or is at risk of stroke or transient ischemic attack (TIA).The subject who has or has had TIA, commonly referred to as " mini-stroke ", is at risk of stroke, particularly the risk of stroke within a few days after TIA.Therefore, in some embodiments, the subject who has experienced or has experienced TIA (for example, within the past 2 days) is administered lipid-binding protein according to the dosage regimen described herein.

[0284] In some embodiments, the subject has or is at risk of having cytokine release syndrome (CRS, cytokine storm). In some embodiments, the CRS is secondary to an infection, e.g., a bacterial or viral infection.

[0285] In some embodiments, the subject has or is at risk for organ transplant, eg, heart transplant rejection.

[0286] In some embodiments, the subject has or is at risk for ischemia-reperfusion induced tissue injury.

[0287] In some embodiments, the subject has or is at risk for post-surgical inflammation.

[0288] In some embodiments, the subject has or is at risk for psoriasis.

[0289] In some embodiments, the subject has or is at risk of sepsis, including, but not limited to, sepsis in which the subject has abnormal levels of at least two of TNFα, IL-6, IL-8, TREM-1, and kynurenine pathway biomarkers, such as abnormal levels of TREM-1, and abnormal levels of at least one of quinolinic acid, kynurenic acid, kynurenine, tryptophan, and the kynurenine / tryptophan ratio. In some embodiments, the sepsis is secondary to an infection, such as a bacterial or viral infection.

[0290] In some embodiments, a subject with sepsis has a documented or suspected infection and an acute change in total SOFA score of 2 or more (see Vincent et al. 1996, Intensive Care Med, 22:707-710).

[0291] In some embodiments, the subject has septic shock.In some embodiments, the subject with septic shock has hypotension (for example, systolic arterial pressure less than 90mmHg or mean arterial pressure (MAP) less than 65mmHg) that requires the use of vasopressors for more than 1 hour despite intravenous fluid resuscitation.In some embodiments, the subject with septic shock has hypotension that requires vasopressor therapy to maintain a mean arterial pressure of 65mmHg or above, and has serum lactate levels greater than 2mmol / L (greater than 18mg / dL) despite adequate fluid resuscitation.

[0292] In some embodiments, the subject has or is at risk for sepsis-induced acute kidney injury (AKI).

[0293] In some embodiments, the subject has or is at risk for hypoalbuminemia.

[0294] In some embodiments, the subject has or is at risk for hypoalbuminemia associated with a vitamin deficiency.

[0295] In some embodiments, the subject has or is at risk for hypoalbuminemia associated with inflammatory bowel disease (IBD).

[0296] In some embodiments, the subject has or is at risk for hypoalbuminemia associated with kidney disease.

[0297] In some embodiments, the subject has or is at risk for hypoalbuminemia associated with an infection.

[0298] In some embodiments, the subject has or is at risk for stress-related hypoalbuminemia.

[0299] In some embodiments, the subject has or is at risk for hypoalbuminemia associated with thyroid disease.

[0300] In some embodiments, the subject has or is at risk for diabetes-related hypoalbuminemia.

[0301] In some embodiments, the subject has or is at risk for hypoalbuminemia associated with nephrotic syndrome.

[0302] In some embodiments, the subject has or is at risk for hypoalbuminemia associated with lupus.

[0303] In some embodiments, the subject has or is at risk for hypoalbuminemia associated with cirrhosis.

[0304] In some embodiments, the subject has or is at risk for hypoalbuminemia associated with liver disease.

[0305] In some embodiments, the subject has or is at risk for hypoalbuminemia associated with heart failure.

[0306] In some embodiments, the subject has or is at risk for hypoalbuminemia associated with malnutrition.

[0307] In some embodiments, the subject has or is at risk for attention-deficit / hyperactivity disorder (ADHD).

[0308] In some embodiments, the subject has or is at risk for a central nervous system (CNS) disease.

[0309] In some embodiments, the subject has or is at risk for COVID-19 cognitive decline.

[0310] In some embodiments, the subject has or is at risk for depression or major depressive disorder.

[0311] In some embodiments, the subject has or is at risk for epilepsy.

[0312] In some embodiments, the subject has or is at risk for an HIV-associated neurocognitive disorder.

[0313] In some embodiments, the subject has or is at risk for Huntington's disease.

[0314] In some embodiments, the subject has or is at risk for inflammatory bowel disease (IBD).

[0315] In some embodiments, the subject has or is at risk for long-term cognitive decline ("brain fog"), such as can develop after sepsis.

[0316] In some embodiments, the subject has or is at risk of death or neurological deficit following cardiac arrest.

[0317] In some embodiments, the subject has or is at risk for multiple sclerosis (MS).

[0318] In some embodiments, the subject has or is at risk for Parkinson's disease.

[0319] In some embodiments, the subject has or is at risk for schizophrenia.

[0320] In some embodiments, the subject has or is at risk for a vascular endothelial disorder.

[0321] In some embodiments, the subject has or is at risk for acute respiratory distress syndrome (ARDS).

[0322] In some embodiments, the subject has a SOFA score of 1 to 24, e.g., a score of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24, prior to treatment with the lipid-binding protein molecule (see Vincent et al. 1996, Intensive Care Med, 22:707-710).

[0323] In some embodiments, the subject has or is at risk for acute kidney injury (AKI), eg, AKI resulting from a viral or bacterial infection (eg, a septic subject).

[0324] In some embodiments, the subject may have CRS, or may be at risk for CRS, and / or may require a reduction in the serum level of one or more inflammatory markers, such as IL-6. In some embodiments, the subject has CRS. In some embodiments, the subject has CRS that is secondary to an infection, such as a viral or bacterial infection. In yet other embodiments, the subject is at risk for CRS, such as CRS caused by a viral or bacterial infection.

[0325] In another embodiment, the subject is a subject who needs to reduce the serum level of one or more inflammatory markers, for example, a subject who has one or more inflammatory marker levels that are elevated 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 chemotactic protein (MCP) 1, triggering receptor expressed on myeloid cells-1 (TREM-1), and tumor necrosis factor alpha (TNFα).In some embodiments, the one or more cytokines include IL-6. In some embodiments, the one or more cytokines include combinations of the above, e.g., combinations of two, three, four, five, six, seven, eight, or all nine 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, TREM-1, and tumor necrosis factor alpha (TNFα).

[0326] 6.3. Dosage Regimen The methods of the present disclosure typically involve multiple administrations of a lipid binding protein molecule (e.g., ApoA-I), e.g., 2 to 20 individual doses, although in some embodiments, a single dose can be used. In some embodiments, the administration regimen can include two or more, three or more, or four or more individual doses of a lipid binding protein molecule (e.g., ApoA-I), e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more than 20 individual doses. In some embodiments, the administration regimen comprises or consists of a single dose. In some embodiments, the administration regimen comprises or consists of two individual doses. In some embodiments, the administration regimen comprises or consists of three individual doses. In some embodiments, the administration regimen comprises or consists of four individual doses. In some embodiments, the administration regimen comprises or consists of five individual doses. In some embodiments, the dosing regimen includes or consists of 6 individual doses. In some embodiments, the dosing regimen includes or consists of 7 individual doses. In some embodiments, the dosing regimen includes or consists of 8 individual doses. In some embodiments, the dosing regimen includes or consists of 9 individual doses. In some embodiments, the dosing regimen includes or consists of 10 individual doses. In some embodiments, the dosing regimen includes or consists of 11 individual doses. In some embodiments, the dosing regimen includes or consists of 12 individual doses. In some embodiments, the dosing regimen includes or consists of 13 individual doses. In some embodiments, the dosing regimen includes or consists of 14 individual doses. In some embodiments, the dosing regimen includes or consists of 15 individual doses. In some embodiments, the dosing regimen includes or consists of 16 individual doses.In some embodiments, the dosing regimen comprises or consists of 17 individual doses, in some embodiments, the dosing regimen comprises or consists of 18 individual doses, in some embodiments, the dosing regimen comprises or consists of 19 individual doses, in some embodiments, the dosing regimen comprises or consists of 20 individual doses.

[0327] In some embodiments, the amount of lipid-binding protein molecule delivered by each dose can be 5 mg / kg to 40 mg / kg on a protein weight basis. In some embodiments, the amount of lipid-binding protein molecule delivered by each dose can be 5 mg / kg to 10 mg / kg on a protein weight basis. In some embodiments, the amount of lipid-binding protein molecule delivered by each dose can be 5 mg / kg to 20 mg / kg on a protein weight basis. In some embodiments, the amount of lipid-binding protein molecule delivered by each dose can be 10 mg / kg to 30 mg / kg on a protein weight basis. In some embodiments, the amount of lipid-binding protein molecule delivered by each dose can be 10 mg / kg to 20 mg / kg on a protein weight basis. In some embodiments, the dose is 5 mg / kg on a protein weight basis. In some embodiments, the dose is 6 mg / kg on a protein weight basis. In some embodiments, the dose is 7 mg / kg on a protein weight basis. In some embodiments, the dose is 8 mg / kg on a protein weight basis. In some embodiments, the dose is 9 mg / kg on a protein weight basis. In some embodiments, the dose is 10 mg / kg by weight protein. In some embodiments, the dose is 11 mg / kg by weight protein. In some embodiments, the dose is 12 mg / kg by weight protein. In some embodiments, the dose is 13 mg / kg by weight protein. In some embodiments, the dose is 14 mg / kg by weight protein. In some embodiments, the dose is 15 mg / kg by weight protein. In some embodiments, the dose is 16 mg / kg by weight protein. In some embodiments, the dose is 17 mg / kg by weight protein. In some embodiments, the dose is 18 mg / kg by weight protein. In some embodiments, the dose is 19 mg / kg by weight protein.In some embodiments, the dose is 20 mg / kg by weight protein. In some embodiments, the dose is 21 mg / kg by weight protein. In some embodiments, the dose is 22 mg / kg by weight protein. In some embodiments, the dose is 23 mg / kg by weight protein. In some embodiments, the dose is 24 mg / kg by weight protein. In some embodiments, the dose is 25 mg / kg by weight protein. In some embodiments, the dose is 26 mg / kg by weight protein. In some embodiments, the dose is 27 mg / kg by weight protein. In some embodiments, the dose is 28 mg / kg by weight protein. In some embodiments, the dose is 29 mg / kg by weight protein. In some embodiments, the dose is 30 mg / kg by weight protein. In some embodiments, the dose is 31 mg / kg by weight protein. In some embodiments, the dose is 32 mg / kg by weight protein. In some embodiments, the dose is 33 mg / kg by weight protein. In some embodiments, the dose is 34 mg / kg by protein weight. In some embodiments, the dose is 35 mg / kg by protein weight. In some embodiments, the dose is 36 mg / kg by protein weight. In some embodiments, the dose is 37 mg / kg by protein weight. In some embodiments, the dose is 38 mg / kg by protein weight. In some embodiments, the dose is 39 mg / kg by protein weight. In some embodiments, the dose is 40 mg / kg by protein weight.

[0328] An exemplary dosing regimen for a subject with sepsis (e.g., septic shock) involves twice-daily administration of a lipid-binding protein molecule (e.g., ApoA-I) for five days. In some embodiments, ApoA-I (e.g., as CER-001) is administered at a dose of 10 mg / kg protein weight. In other embodiments, ApoA-I (e.g., as CER-001) is administered at a dose of 20 mg / kg protein weight. The two daily doses can be administered, for example, approximately 12 hours apart. In some embodiments, the two daily doses are administered 11 to 13 hours apart. In some embodiments, the two daily doses are administered as a morning dose and an evening dose, which can be more or less than 12 hours apart.

[0329] In some embodiments of the dosing regimens described in this paragraph, the subject has septic shock.

[0330] In some embodiments, the lipid-binding protein molecule is administered according to an induction regimen, and optionally according to a consolidation regimen, where the induction and consolidation regimens are described in Sections 6.3.1 and 6.3.2, respectively. In some embodiments, the lipid-binding protein molecule can be administered in a single phase, for example, according to the administration regimens described in this section. In some embodiments, the subject is not treated with the lipid-binding protein molecule according to a maintenance regimen, for example, a regimen comprising long-term (e.g., one month or more) administration of the lipid-binding protein molecule. In other embodiments, for example, when the subject has a chronic condition, such as Alzheimer's disease, the subject is treated with the lipid-binding protein molecule according to a maintenance regimen, for example, a regimen comprising long-term (e.g., one month or more) administration of the lipid-binding protein molecule.

[0331] A lipid binding protein molecule (eg, ApoA-I) administration regimen of the present disclosure can last up to one week, one week, or more than one week (eg, two or three weeks).

[0332] For example, a lipid binding protein molecule (e.g., ApoA-I) administration regimen may include: Five individual doses of ApoA-I administered over a period of one week Six individual doses of ApoA-I administered over a period of one week Seven individual doses of ApoA-I over a period of one week Eight individual doses of ApoA-I administered over a period of one week - 9 individual doses of ApoA-I over a period of 1 week - 10 individual doses of ApoA-I over a period of 1 week - 12 individual doses of ApoA-I over a period of 1 week - 14 individual doses of ApoA-I over a period of 1 week 10 individual doses of ApoA-I over 2 weeks 12 individual doses of ApoA-I over 2 weeks - It may include 14 individual doses of ApoA-I administered over a two week period.

[0333] In one embodiment, the method of the disclosure involves administering seven individual doses of ApoA-I over a period of one week, for example, on days 1, 2, 3, 4, 5, 6, and 7.

[0334] In some embodiments, the methods of the present disclosure include administering multiple individual doses over a period of 4 to 6 days, e.g., administering 1 to 2 individual doses per day for 4 to 6 days. In some embodiments, the methods of the present disclosure include administering multiple individual doses over a period of 4 days, e.g., administering 1 to 2 individual doses per day for 4 days. In some embodiments, the methods of the present disclosure include administering multiple individual doses over a period of 5 days, e.g., administering 1 to 2 individual doses per day for 5 days. In some embodiments, the methods of the present disclosure include administering multiple individual doses over a period of 6 days, e.g., administering 1 to 2 individual doses per day for 6 days. In some embodiments of the methods described in this paragraph, one individual dose is administered per day. In other embodiments of the methods described in this paragraph, two individual doses are administered per day.

[0335] In some embodiments, of the methods of the present disclosure, multiple doses of a lipid-binding protein molecule (e.g., ApoA-I) are administered at intervals of one day or less. For example, in some embodiments, two or more individual doses are administered approximately 12 hours apart. In some embodiments, two individual doses are administered approximately 12 hours apart. In other embodiments, three individual doses are administered approximately 12 hours apart. In other embodiments, two individual doses are administered approximately 12 hours apart, with a third individual dose administered approximately one day later. In other embodiments, three individual doses are administered approximately 12 hours apart, with a fourth individual dose administered approximately one day later.

[0336] In some embodiments of the disclosed methods, a lipid binding protein molecule (e.g., ApoA-I) is administered to a subject at 0 and 12 hours (e.g., over a 0.5-1 hour period), e.g., at a dose of 10 mg / kg or 15 mg / kg. In some embodiments of the disclosed methods, a lipid binding protein molecule (e.g., ApoA-I) is administered to a subject at 0 and 12, 24, and 48 hours (e.g., over a 0.5-1 hour period), e.g., at a dose of 10 mg / kg or 15 mg / kg.

[0337] In some embodiments of the disclosed methods, the lipid-binding protein molecule (e.g., ApoA-I) is administered daily, for example, for at least 5 days, at least 6 days, at least 7 days, or more than 7 days (e.g., daily for up to 1 week, or daily for up to 2 weeks). In other embodiments, the lipid-binding protein molecule (e.g., ApoA-I) is administered less frequently, for example, every other day, twice per week, three times per week, or once per week.

[0338] In practice, a dosing window can be provided to accommodate slight variations to a dosing schedule, for example, multiple dosing per week. For example, a window of ±2 days or ±1 day around the dosing date can be used.

[0339] The lipid binding protein molecule (e.g., ApoA-I) can be administered in the methods of the present disclosure for a predetermined period of time, for example, one week. Alternatively, administration of the lipid binding protein molecule (e.g., ApoA-I) can be continued until one or more symptoms of the condition are reduced, or until the serum levels of one or more inflammatory markers are reduced, for example, reduced to normal levels or reduced relative to a baseline value for the subject, for example, a baseline value measured before initiation of lipid binding protein molecule (e.g., ApoA-I) therapy. Reference or "normal" levels for various markers, e.g., inflammatory markers, are known in the art. For example, the Mayo Clinic Laboratories Test Catalog (www.mayocliniclabs.com / test-catalog) provides the following reference values: IL-6: 1.8 pg / ml or less; C-reactive protein: 8.0 mg / ml or less; D-dimer: Fibrinogen Equivalent Units (FEU); The following reference values ​​are provided: IFN-γ (FEU) less than 500 ng / mL; ferritin: 24-336 mcg / L (men) and 11-307 mcg / L (women); IL-8 less than 57.8 pg / mL; TNF-α less than 5.6 pg / mL; albumin: 3.5-5.0 g / dL; tryptophan: 17-80 nmol / mL; and serotonin less than 330 ng / mL. Radhakrishnan et al., 2014, Indian J Endocrinol Metab 18(4):505-510, provide the following reference values: MCP-1: 350-450 pg / mL. Ho et al., 2004, World J Gastroenterol 10(14):2014-2018, provide the following reference values: VCAM-1: less than 700 ng / mL. Rothlein et al., 1991, J Immunol. 147(11):3788-93 provides the following reference values: ICAM-1: 100-200 ng / mL. The normal range for TREM-1 is estimated to be less than 500 pg / mL, and for some subject populations, it may be less than 300 pg / mL. Values ​​outside the normal range are considered "abnormal."

[0340] The disclosed methods typically involve administering a high dose of a lipid-binding protein molecule (e.g., ApoA-I). 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), which are administered, for example, over one or more days (e.g., a one-day period, a two-day period, a three-day period, a four-day period, a five-day period, a six-day period, a seven-day period, a eight-day period, a nine-day period, a ten-day period, an eleven-day period, a twelve-day period, a thirteen-day period, a fourteen-day period, or a fifteen-day period). In some embodiments, the individual doses of the high dose are administered daily, twice daily, or at intervals of 2 to 3 days.

[0341] In some embodiments, the high dose is an amount effective to increase a subject's HDL and / or ApoA-I blood levels and / or improve a subject's vascular endothelial function, as measured, for example, by circulating vascular cell adhesion molecule 1 (VCAM-1) and / or intercellular adhesion molecule 1 (ICAM-1) levels. In some embodiments, the high dose or individual doses are amounts that increase a subject's HDL and / or ApoA-I levels by at least 25%, at least 30%, or at least 35% 2-4 hours after administration.

[0342] In some embodiments, the high dose is an amount effective to reduce serum levels of one or more inflammatory markers, such as one or more of IL-6, C-reactive protein, D-dimer, ferritin, IL-8, GM-CSF, MCP1, TREM-1, or TNF-α. In some embodiments, the serum levels of one or more inflammatory markers are reduced from an elevated range to the normal range and / or are reduced by at least 20%, at least 40%, or at least 60%.

[0343] In some embodiments, the dose of a lipid-binding protein molecule (e.g., ApoA-I) administered to a subject (e.g., an individual dose that forms a higher dose when aggregated with one or more other individual doses) can be in the range of 4 to 40 mg / kg (e.g., 10 to 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 above values, e.g., 10 to 20 mg / kg, 15 to 25 mg / kg, 20 to 40 mg / kg, 25 to 35 mg / kg, or 30 to 40 mg / kg). As used herein, the phrase "protein weight basis" means that the dose of a lipid-binding protein molecule (e.g., ApoA-I) administered to a subject is calculated based on the amount of lipid-binding protein molecule (e.g., ApoA-I) administered and the subject's body weight. For example, a subject weighing 70 kg receiving a 20 mg / kg dose of CER-001 will receive an amount of CER-001 that provides 1400 mg of ApoA-I (70 kg x 20 mg / kg).

[0344] The same amount of lipid-binding protein can be administered for each individual dose. Alternatively, the amount of lipid-binding protein can vary between individual doses. For example, one or more individual doses can be administered with the amount of a first dose, and then one or more individual doses can be administered with a different amount of a second dose. For example, the amount of the first dose can be 5 mg / kg (protein weight basis), and the amount of the second dose can be a higher amount, for example, 10 mg / kg (protein weight basis). As another example, the amount of the first dose can be 10 mg / kg (protein weight basis), and the amount of the second dose can be a higher amount, for example, 15 mg / kg (protein weight basis). As another example, the amount of the first dose can be 10 mg / kg (protein weight basis), and the amount of the second dose can be a higher amount, for example, 20 mg / kg (protein weight basis). As another example, the amount of the first dose can be 15 mg / kg (protein weight basis), and the amount of the second dose can be a higher amount, for example, 20 mg / kg (protein weight basis). As another example, the amount of the first dose can be 20 mg / kg (protein weight basis), and the amount of the second dose can be a lower amount, for example, 10 mg / kg (protein weight basis). As another example, the amount of the first dose can be 20 mg / kg (protein weight basis), and the amount of the second dose can be a lower amount, for example, 5 mg / kg (protein weight basis). As another example, the amount of the first dose can be 10 mg / kg (protein weight basis), and the amount of the second dose can be a lower amount, for example, 5 mg / kg (protein weight basis).

[0345] In some embodiments, the lipid-binding protein (e.g., ApoA-I, optionally in the form of CER-001) is administered at a dose of 5 mg / kg to 20 mg / kg (protein weight basis) once or twice daily for 4 to 6 days. In some embodiments, the amount of each individual dose is the same; alternatively, different individual dose amounts may be used, e.g., as described in the preceding paragraph. In some embodiments, the lipid-binding protein (e.g., ApoA-I, optionally in the form of CER-001) is administered at a dose of 5 mg / kg to 20 mg / kg (protein weight basis) once or twice daily for 4 days. In some embodiments, the lipid-binding protein (e.g., ApoA-I, optionally in the form of CER-001) is administered at a dose of 5 mg / kg to 20 mg / kg (protein weight basis) once or twice daily for 5 days. In some embodiments, the lipid-binding protein (e.g., ApoA-I, optionally in the form of CER-001) is administered at a dose of 5 mg / kg to 20 mg / kg (by protein weight) once or twice daily for 6 days. In some embodiments, the lipid-binding protein (e.g., ApoA-I, optionally in the form of CER-001) is administered at a dose of 5 mg / kg (by protein weight) once or twice daily for 4 to 6 days. In some embodiments, the lipid-binding protein (e.g., ApoA-I, optionally in the form of CER-001) is administered at a dose of 5 mg / kg (by protein weight) once or twice daily for 4 days. In some embodiments, the lipid-binding protein (e.g., ApoA-I, optionally in the form of CER-001) is administered at a dose of 5 mg / kg (by protein weight) once or twice daily for 5 days. In some embodiments, the lipid-binding protein (e.g., ApoA-I, optionally in the form of CER-001) is administered at a dose of 5 mg / kg (protein weight basis) once or twice daily for 6 days. In some embodiments, the lipid-binding protein (e.g., ApoA-I, optionally in the form of CER-001) is administered at a dose of 10 mg / kg (protein weight basis) once or twice daily for 4 to 6 days.In some embodiments, the lipid-binding protein (e.g., ApoA-I, optionally in the form of CER-001) is administered at a dose of 10 mg / kg (protein weight basis) once or twice daily for 4 days. In some embodiments, the lipid-binding protein (e.g., ApoA-I, optionally in the form of CER-001) is administered at a dose of 10 mg / kg (protein weight basis) once or twice daily for 5 days. In some embodiments, the lipid-binding protein (e.g., ApoA-I, optionally in the form of CER-001) is administered at a dose of 10 mg / kg (protein weight basis) once or twice daily for 6 days. In some embodiments, the lipid-binding protein (e.g., ApoA-I, optionally in the form of CER-001) is administered at a dose of 15 mg / kg (protein weight basis) once or twice daily for 4 to 6 days. In some embodiments, the lipid-binding protein (e.g., ApoA-I, optionally in the form of CER-001) is administered at a dose of 15 mg / kg (protein weight basis) once or twice daily for 4 days. In some embodiments, the lipid-binding protein (e.g., ApoA-I, optionally in the form of CER-001) is administered at a dose of 15 mg / kg (protein weight basis) once or twice daily for 5 days. In some embodiments, the lipid-binding protein (e.g., ApoA-I, optionally in the form of CER-001) is administered at a dose of 15 mg / kg (protein weight basis) once or twice daily for 6 days. In some embodiments, the lipid-binding protein (e.g., ApoA-I, optionally in the form of CER-001) is administered at a dose of 20 mg / kg (protein weight basis) once or twice daily for 4 to 6 days. In some embodiments, the lipid binding protein (e.g., ApoA-I, optionally in the form of CER-001) is administered at a dose of 20 mg / kg (protein weight basis) once or twice daily for 4 days. In some embodiments, the lipid binding protein (e.g., ApoA-I, optionally in the form of CER-001) is administered at a dose of 20 mg / kg (protein weight basis) once or twice daily for 5 days.In some embodiments, the lipid-binding protein (e.g., ApoA-I, optionally in the form of CER-001) is administered at a dose of 20 mg / kg (on a protein weight basis) once or twice daily for 6 days. In some embodiments of the methods described in this paragraph, the lipid-binding protein is administered once daily. In other embodiments of the methods described in this paragraph, the lipid-binding protein is administered twice daily.

[0346] In some embodiments, the lipid binding protein (eg, ApoA-I, optionally in the form of CER-001) is administered to a subject according to one of the following regimens. - two individual doses per day at a first dose amount (e.g., 5 mg / kg, 10 mg / kg, 15 mg / kg, or 20 mg / kg) for at least three days, followed by one individual dose per day at a second dose amount equal to or higher than the first dose amount (e.g., 5 mg / kg, 10 mg / kg, 15 mg / kg, or 20 mg / kg) for up to 15 days (e.g., up to 5 days, up to 1 week, up to 10 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, or 15 days); - two individual doses per day at a first dose amount (e.g., 5 mg / kg, 10 mg / kg, 15 mg / kg, or 20 mg / kg) for three days, followed by one individual dose per day at a second dose amount equal to or higher than the first dose amount (e.g., 5 mg / kg, 10 mg / kg, 15 mg / kg, or 20 mg / kg) for up to 15 days (e.g., up to 5 days, up to 1 week, up to 10 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, or 15 days); - two individual doses per day at a first dose amount (e.g., 5 mg / kg, 10 mg / kg, 15 mg / kg, or 20 mg / kg) for four days, followed by one individual dose per day at a second dose amount equal to or higher than the first dose amount (e.g., 5 mg / kg, 10 mg / kg, 15 mg / kg, or 20 mg / kg) for up to 15 days (e.g., up to 5 days, up to 1 week, up to 10 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, or 15 days); - two individual doses per day at a first dose amount (e.g., 5 mg / kg, 10 mg / kg, 15 mg / kg, or 20 mg / kg) for 5 days, followed by one individual dose per day at a second dose amount equal to or higher than the first dose amount (e.g., 5 mg / kg, 10 mg / kg, 15 mg / kg, or 20 mg / kg) for up to 15 days (e.g., up to 5 days, up to 1 week, up to 10 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, or 15 days); - Two individual doses per day at a first dose amount (e.g., 5 mg / kg, 10 mg / kg 15 mg / kg, or 20 mg / kg) for 6 days, followed by one individual dose per day at a second dose amount equal to or higher than the first dose amount (e.g., 5 mg / kg, 10 mg / kg 15 mg / kg, or 20 mg / kg) for up to 15 days (e.g., up to 5 days, up to 1 week, up to 10 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, or 15 days). In some embodiments, the amount of the first dose is the same as the amount of the second dose. For example, in some embodiments, the amounts of the first and second doses are 5 mg / kg, 10 mg / kg, 15 mg / kg, or 20 mg / kg. In other embodiments, the amount of the second dose is higher than the amount of the first dose. For example, in some embodiments, the amount of the first dose is 5 mg / kg and the amount of the second dose is 10 mg / kg, 15 mg / kg, or 20 mg / kg; the amount of the first dose is 10 mg / kg and the amount of the second dose is 15 mg / kg or 20 mg / kg; or the amount of the first dose is 15 mg / kg and the amount of the second dose is 20 mg / kg.

[0347] In still other embodiments, the lipid-binding protein molecule (e.g., ApoA-I) can be administered on a unit dosage basis. The unit dosage used in the methods of the present disclosure can, in some embodiments, range from 300 mg to 4000 mg (e.g., 600 mg to 4000 mg) (protein weight basis) per administration.

[0348] In certain embodiments, the dosage of the lipid binding protein molecule (e.g., ApoA-I) per administration is 300 mg to 400 mg, 300 mg to 500 mg, 300 mg to 600 mg, 300 mg to 800 mg, 300 mg to 1000 mg, 300 mg to 1200 mg, 300 mg to 1500 mg, 300 mg to 2000 mg, 300 mg to 2400 mg, 300 mg to 3000 mg, 400 mg to 500 mg, 400 mg to 600 mg, 400 mg to 800 mg, 400 mg to 1000 mg, 400 mg to 1 200mg, 400mg~1500mg, 400mg~2000mg, 400mg~2400mg, 400mg~3000mg, 400mg~4000mg, 500mg~600mg, 500mg~800mg, 500mg~1000mg, 500mg~1 200mg, 500mg~1500mg, 500mg~2000mg, 500mg~2400mg, 500mg~3000mg, 500mg~4000mg, 600mg~800mg, 600mg~1000mg, 600mg~1200mg, 600mg~1 500mg, 600mg~2000mg, 600mg~2400mg, 600mg~3000mg, 600mg~4000mg, 800mg~1000mg, 800mg~1200mg, 800mg~1500mg, 800mg~2000mg, 800mg ~2400mg, 800mg~3000mg, 800mg~4000mg, 1000mg~1200mg, 1000mg~1500mg, 1000mg~2000mg, 1000mg~2400mg, 1000mg~3000mg, 1000mg~4000m g, 1200mg to 1500mg, 1200mg to 2000mg, 1200mg to 2400mg, 1200mg to 3000mg, 1200mg to 4000mg, 1500mg to 2000mg, 1500mg to 2400mg, 1500mg to 3000mg, 1500mg to 4000mg, 2000mg to 2400mg, 2000mg to 3000mg, 2000mg to 4000mg, 2400mg to 3000mg, 2400mg to 4000mg, or 3000mg to 4000mg (on a protein weight basis).

[0349] In some embodiments, the high dose of lipid-binding protein molecule (e.g., ApoA-I), e.g., the sum of multiple individual doses, is 600 mg to 40 g (protein weight basis). In certain embodiments, the high dose is 3 g to 35 g or 5 g to 30 g (protein weight basis).

[0350] The lipid-binding protein molecule (e.g., ApoA-I) is preferably administered as an IV infusion. For example, a stock solution of CER-001 can be diluted in normal saline, e.g., physiological saline (0.9% NaCl), to a total volume of 125 to 250 ml. In some embodiments, subjects weighing less than 80 kg have a total volume of 125 ml, and subjects weighing at least 80 kg have a total volume of 250 ml. In some embodiments, a dose of CER-001 is administered in a total volume of 250 ml. The lipid-binding protein molecule (e.g., ApoA-I) can be administered over a period ranging from 1 hour to 24 hours. Depending on the needs of the subject, administration can be by slow infusion having a duration of more than 1 hour (e.g., up to 2 hours or up to 24 hours), by rapid infusion of 1 hour or less, or by a single bolus injection. In one embodiment, the lipid-binding protein molecule (e.g., ApoA-I) is administered using an infusion pump at a fixed rate of, for example, 125 ml / hour or 250 ml / hour over a period of 1 hour. In one embodiment, the dose of lipid-binding protein molecule (e.g., ApoA-I) is administered as an infusion over a 24-hour period.

[0351] 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-binding protein molecule (e.g., ApoA-I) over multiple consecutive days, for example, 3 consecutive days, 4 consecutive days, 5 consecutive days, or 6 consecutive days. In some embodiments, the lipid-binding protein molecule (e.g., ApoA-I) is administered for 4 consecutive days. In some embodiments, the lipid-binding protein molecule (e.g., ApoA-I) is administered for 5 consecutive days. In some embodiments, the lipid-binding protein molecule (e.g., ApoA-I) is administered for 6 consecutive days.

[0352] In some embodiments, induction regimens suitable for use in the methods of the present disclosure involve twice-daily administration of a lipid-binding protein molecule (e.g., ApoA-I), e.g., twice-daily administration on multiple consecutive days. Twice-daily administration can include, for example, two doses approximately 12 hours apart, or a morning dose and an evening dose, which can be more or less than 12 hours apart.

[0353] In one embodiment, the induction regimen comprises two doses of a lipid-binding protein molecule (e.g., ApoA-I) per day for three consecutive days. In one embodiment, the induction regimen comprises two doses of a lipid-binding protein molecule (e.g., ApoA-I) per day for four consecutive days. In one embodiment, the induction regimen comprises two doses of a lipid-binding protein molecule (e.g., ApoA-I) per day for five consecutive days. In one embodiment, the induction regimen comprises two doses of a lipid-binding protein molecule (e.g., ApoA-I) per day for six consecutive days.

[0354] The therapeutic dose of the lipid-binding protein molecule (e.g., ApoA-I) administered by infusion in the induction regimen can be in the range of 4 to 40 mg / kg (e.g., 4 to 30 mg / kg) (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 above values, e.g., 5 to 15 mg / kg, 10 to 20 mg / kg, or 15 to 25 mg / kg) on ​​a protein weight basis. In some embodiments, the dose of the lipid-binding protein molecule (e.g., ApoA-I) used in the induction regimen is 5 mg / kg. In some embodiments, the dose of the lipid-binding protein molecule (e.g., ApoA-I) used in the induction regimen is 10 mg / kg. In some embodiments, the dose of the lipid-binding protein molecule (e.g., ApoA-I) used in the induction regimen is 15 mg / kg. In some embodiments, the dosage of the lipid-binding protein molecule (e.g., ApoA-I) used in the induction regimen is 20 mg / kg. In some embodiments, the induction regimen comprises six doses of the lipid-binding protein molecule (e.g., ApoA-I) administered over three days at doses of 5 mg / kg, 10 mg / kg, 15 mg / kg, or 20 mg / kg. In some embodiments, the induction regimen comprises eight doses of the lipid-binding protein molecule (e.g., ApoA-I) administered over four days at doses of 5 mg / kg, 10 mg / kg, 15 mg / kg, or 20 mg / kg. In some embodiments, the induction regimen comprises ten doses of the lipid-binding protein molecule (e.g., ApoA-I) administered over five days at doses of 5 mg / kg, 10 mg / kg, 15 mg / kg, or 20 mg / kg. In some embodiments, the induction regimen comprises 12 doses of a lipid binding protein molecule (e.g., ApoA-I) administered over 6 days at doses of 5 mg / kg, 10 mg / kg, 15 mg / kg, or 20 mg / kg.

[0355] In yet other embodiments, the lipid binding protein molecule (e.g., ApoA-I) can be administered on a unit dosage basis. The unit dosage used in the induction phase can, in some embodiments, range from 300 mg to 4000 mg (e.g., 300 mg to 3000 mg) (on a protein weight basis) per administration by infusion.

[0356] In certain embodiments, the dosage of lipid binding protein molecule (e.g., ApoA-I) 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 (on a protein weight basis) per administration by infusion.

[0357] 6.3.2. Intensive regimen Consolidation regimens suitable for use in the methods of the present disclosure involve administering one or more doses of a lipid binding protein molecule (eg, ApoA-I) following an induction regimen.

[0358] In one embodiment, the intensive regimen involves administering two doses of a lipid-binding protein molecule (e.g., ApoA-I). For example, the two doses can be administered approximately 12 hours apart, or as a morning dose and an evening dose (which can be more or less than 12 hours apart).

[0359] In some embodiments, the dose(s) of the lipid binding protein molecule (e.g., ApoA-I) in the intensification regimen can be administered on day 6 of a dosing regimen that begins with an induction regimen on day 1. In some embodiments, the dose(s) of the lipid binding protein molecule (e.g., ApoA-I) in the intensification regimen can be administered on day 4 of a dosing regimen that begins with an induction regimen on day 1. In some embodiments, the dose(s) of the lipid binding protein molecule (e.g., ApoA-I) in the intensification regimen can be administered on day 5 of a dosing regimen that begins with an induction regimen on day 1. In some embodiments, the dose(s) of the lipid binding protein molecule (e.g., ApoA-I) in the intensification regimen can be administered on day 7 of a dosing regimen that begins with an induction regimen on day 1.

[0360] In some embodiments, the intensification regimen comprises a once-daily administration of a lipid-binding protein molecule (e.g., ApoA-I) following the induction regimen, and the induction regimen comprises a twice-daily administration of a lipid-binding protein molecule (e.g., ApoA-I). Each individual dose of the intensification regimen can be the same as or higher than each individual dose of the induction regimen. For example, following an induction regimen comprising twice-daily administration of a lipid-binding protein molecule (e.g., ApoA-I) for at least 3 days (e.g., up to 1 week, 3 days, 4 days, 5 days, 6 days, or 7 days), the lipid-binding protein molecule (e.g., ApoA-I) can be administered once a day for up to 15 days (e.g., up to 7 days, up to 10 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, or 15 days).

[0361] The therapeutic dose of the lipid-binding protein molecule (e.g., ApoA-I) administered by infusion in the intensive regimen can be in the range of 4 mg / kg to 40 mg / kg (e.g., 4 to 30 mg / kg) (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 above values, e.g., 5 to 15 mg / kg, 10 to 20 mg / kg, or 15 to 25 mg / kg) on ​​a protein weight basis. In some embodiments, the dose of the lipid-binding protein molecule (e.g., ApoA-I) used in the intensive regimen is 5 mg / kg. In some embodiments, the dose of the lipid-binding protein molecule (e.g., ApoA-I) used in the intensive regimen is 10 mg / kg. In some embodiments, the dose of the lipid-binding protein molecule (e.g., ApoA-I) in the intensive regimen is 15 mg / kg. In some embodiments, the dose of lipid-binding protein molecule (e.g., ApoA-I) used in the intensive regimen is 20 mg / kg. In some embodiments, the intensive regimen includes two doses of lipid-binding protein molecule (e.g., ApoA-I) administered daily at doses of 5 mg / kg, 10 mg / kg, 15 mg / kg, or 20 mg / kg.

[0362] In yet other embodiments, the lipid binding protein molecule (e.g., ApoA-I) can be administered on a unit dosage basis. The unit dosage used in the consolidation phase can, in some embodiments, range from 300 mg to 4000 mg (e.g., 300 mg to 3000 mg) (on a protein weight basis) per administration by infusion.

[0363] In certain embodiments, the dosage of lipid binding protein molecule (e.g., ApoA-I) used during the consolidation phase is 300 mg to 1500 mg, 400 mg to 1500 mg, 500 mg to 1200 mg, or 500 mg to 1000 mg (on a protein weight basis) per administration by infusion.

[0364] The lipid binding protein molecule (eg, ApoA-I) can be administered during the consolidation phase in the same manner as described in Section 6.3, for example, as an IV infusion over 1 hour.

[0365] Combination Therapy Lipid binding protein molecule (for example, ApoA-I) can be administered to the subject described herein as a single therapy or as part of a combined therapy regimen.For example, combined therapy can include lipid binding protein molecule (for example, ApoA-I) in combination with the standard treatment for the condition that the subject suffers from or is at risk of suffering from, such as sepsis and / or AKI.For example, see Rhodes et al., 2017, Intensive Care Med 43:304-377; Dugar et al., 2020, Cleveland Clinic Journal of Medicine 87(1):53-64; Singer et al., 2016, JAMA 315(8):801-810.

[0366] In some embodiments, for example, for a subject with sepsis, the subject is treated with a lipid-binding protein molecule (e.g., ApoA-I) in combination with fluid replacement therapy. In some embodiments, for example, for a subject with sepsis, the subject is treated with a lipid-binding protein molecule (e.g., ApoA-I) in combination with an antibacterial agent. In some embodiments, for example, for a subject with sepsis, the subject is treated with a lipid-binding protein molecule (e.g., ApoA-I) 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, for example, for a subject with a viral infection, the subject is treated with a lipid-binding protein molecule (e.g., ApoA-I) in combination with an antiviral agent. In some embodiments, the subject is treated with a lipid-binding protein molecule (e.g., ApoA-I) in combination with a medication that increases blood pressure (e.g., norepinephrine or epinephrine). In some embodiments, the subject is treated with a lipid-binding protein molecule (e.g., ApoA-I) in combination with an immunosuppressant, e.g., tacrolimus or everolimus.

[0367] In some embodiments, the combination therapy regimen can include one or more anti-IL-6 agents and / or one or more other agents for treating CRS, such as corticosteroids (e.g., methylprednisolone and / or dexamethasone). Exemplary anti-IL-6 agents include tocilizumab, siltuximab, olokizumab, elcilimomab, BMS-945429, sirukumab, revilimab, and CPSI-2364. In some embodiments, a lipid-binding protein molecule (e.g., ApoA-I) is administered in combination with tocilizumab.

[0368] In certain embodiments, an antihistamine (e.g., dexchlorpheniramine, cetirizine, fexofenadine, or loratadine) can be administered prior to administration of a lipid-binding protein molecule (e.g., ApoA-I). The antihistamine can reduce the likelihood of an allergic reaction. [Example]

[0369] 7. Working Example 7.1. Example 1: Lipid-binding protein molecular therapy in a porcine model of LPS-induced acute kidney injury The ability of CER-001, an ApoA-I-containing complex, to attenuate sepsis-associated cognitive decline was evaluated in a lipopolysaccharide (LPS)-induced porcine model of cognitive decline ("brain fog").

[0370] 7.1.1 Materials and Methods Pigs were randomized into three groups: LPS (endotoxemic pigs, n=3 unless otherwise noted), single-dose CER-001-treated pigs (endotoxemic pigs treated with a single dose of CER-001 at 20 mg / kg, n=4 unless otherwise noted), and multiple-dose CER-001-treated pigs (endotoxemic pigs treated with two doses of CER-001 at 20 mg / kg for a total dose of 40 mg / kg, n=6 unless otherwise noted).

[0371] 7.1.1.1. Kynurenine Pathway Biomarker Assays Sepsis was induced in pigs at T0 by intravenous infusion of saline solution containing 300 μg / kg LPS. Pigs treated with a single dose of CER-001 and pigs treated with multiple doses of CER-001 received a 20 mg / kg dose of CER-001 at T0. Pigs treated with multiple doses of CER-001 received a second 20 mg / kg dose of CER-001 3 hours later (T3). Quinolinic acid, kynurenic acid, tryptophan (Trp), and kynurenine (Kyn) levels, as well as the Kyn:Trp ratio, were monitored over time. LPS pigs, n=3 except where noted; single-dose endotoxemic pigs treated with a single dose of CER-001 at 20 mg / kg, n=4 except where noted; endotoxemic pigs treated with two doses of CER-001 at 20 mg / kg, n=6 except where noted.

[0372] 7.1.1.2. Q-PCR of Kynurenine Pathway and Interleukin-6 mRNA in Brain Tissue For all groups, n = 3. Animals were humanely sacrificed. Brain tissue was extracted, and qPCR was performed on mRNAs encoding aromatic L-amino acid / L-tryptophan decarboxylase (DDC), indoleamine 2,3-dioxygenase 1 (IDO1), interleukin-6 (IL-6), kynurenine 3-monooxygenase (KMO), kynurenine formamidase isoform X1 (AFMID), and kynurenine-oxoglutarate transaminase 3 (KYAT3).

[0373] 7.1.2.Results LPS injection resulted in a time-dependent increase in quinolinic acid in endotoxemic animals compared to basal conditions (T0) (Figure 1A, Figure 1B). CER-001 treatment was able to reverse the LPS effect, as indicated by essentially unchanged quinolinic acid levels, at both a single dose of 20 mg / kg (Figure 1A, "20 mg") and two doses of 20 mg / kg each, totaling 40 mg / kg (Figure 1B, "40 mg"). One of three pigs in the 20 mg / kg group had a time-dependent increase in quinolinic acid at T3 and T6. All 10 pigs in the experimental group had essentially baseline quinolinic acid levels at the end of the experiment. Figure 1C graphically summarizes the results observed for the three groups. *, p<0.05 vs. LPS.

[0374] LPS injection also resulted in a time-dependent increase in kynurenic acid in two of the three endotoxemic animals (Figures 2A and 2B) compared to basal conditions (T0). CER-001 treatment was able to reverse the LPS effect, as indicated by essentially unchanged kynurenic acid levels, at both a single dose of 20 mg / kg (Figure 2A, "20 mg") and two doses of 20 mg / kg each, totaling 40 mg / kg (Figure 2B, "40 mg"). All 10 pigs in the experimental group had essentially baseline kynurenic acid levels at the end of the experiment. Figure 2C graphically summarizes the results observed for the three groups. *, p<0.05 for the 40 mg group versus LPS.

[0375] LPS injection also resulted in a time-dependent decrease in Trp in endotoxemic animals compared to basal conditions (T0) (Figures 3A and 3B). CER-001 treatment was able to reverse the LPS effect, as indicated by either no decrease or an increase in Trp levels, for both the 20 mg / kg group (Figure 3A, "20 mg") and the 2 × 20 mg / kg group (Figure 3B, "40 mg"). All 10 pigs in the experimental group had Trp levels essentially at or above baseline at the end of the experiment. Figure 3C graphically summarizes the results observed for the three groups. *, p<0.05 for the 20 mg group vs. LPS; **, p<0.-05 for the 40 mg group vs. LPS.

[0376] LPS injection also resulted in a time-dependent increase in Kyn in endotoxemic animals compared to basal conditions (T0) (Figures 4A and 4B). CER-001 treatment was able to reverse the LPS effect, as indicated by essentially unchanged Kyn levels for some pigs in both the 20 mg / kg group (Figure 4A, "20 mg," one of two animals) and the 2 × 20 mg / kg group (Figure 4B, "40 mg," five of six animals). In total, six of the eight pigs in the experimental groups in which Kyn levels were measured had essentially baseline Kyn levels at the end of the experiment. Figure 4C graphically summarizes the results observed for the three groups. *, p<0.05 for the 20 mg group versus the 40 mg group; **, p<0.005 versus LPS.

[0377] Figure 5A summarizes the Kyn:Trp ratios observed for the three groups. *, p<0.05 for the 20 mg group versus the 40 mg group; **, p<0.005 versus LPS. Figure 5B summarizes the Kyn:Trp ratios observed for the three groups (LPS, n=5; 20 mg, n=6; 40 mg, n=3) in the second set of pigs.

[0378] Figure 6A shows the fold gene expression (2 fold) of indoleamine 2,3-dioxygenase 1 (IDO1) in brain tissue relative to housekeeping genes as measured by qPCR for cohorts from all three groups of endotoxemic pigs receiving LPS, 20 mg, and 40 mg. -ΔΔCt ) are indicated. IDO1 catalyzes the conversion of tryptophan to formylkynurenine. In brain tissue from the LPS cohort, approximately two-fold greater IDO1 expression was found relative to housekeeping genes compared to essentially unchanged IDO1 expression in the 20 mg and 40 mg CER-001 cohorts. *, p<0.05 for the 20 mg and 40 mg groups relative to LPS.

[0379] Figure 6B shows the relative fold gene expression of aromatic L-amino acid / L-tryptophan decarboxylase (DDC) in brain tissue as measured by qPCR for cohorts from all three groups of endotoxemic pigs: LPS, 20 mg, and 40 mg. DDC catalyzes the conversion of tryptophan to serotonin. CER-001 resulted in increased expression of DDC in both test groups. *, p<0.05 for the 40 mg group versus LPS.

[0380] Figure 6C shows the relative fold gene expression of kynurenine formamidase isoform X1 (AFMID) in brain tissue as measured by qPCR for cohorts from all three groups of endotoxemic pigs: 20 mg LPS, 40 mg LPS, and 20 mg LPS. AFMID catalyzes the conversion of formylkynurenine to kynurenine. CER-001 partially reversed the increase in AFMID expression induced by LPS.

[0381] Figure 6D shows the relative fold gene expression of kynurenine 3-monooxygenase (KMO) in brain tissue as measured by qPCR for cohorts from all three groups of endotoxemic pigs: 20 mg LPS, and 40 mg LPS. KMO catalyzes the conversion of kynurenine to 3-hydroxykynurenine. CER-001 partially reversed the increase in KMO expression induced by LPS. *, p<0.05 for the 40 mg group versus LPS.

[0382] Figure 6E shows the relative fold gene expression of kynurenine-oxoglutarate transaminase 3 (KYAT3) in brain tissue as measured by qPCR for cohorts from all three groups of endotoxemic pigs treated with LPS (20 mg, 40 mg, and 20 mg). KYAT3 catalyzes the conversion of 3-hydroxykynurenine to xanthurenic acid. CER-001 partially reversed the increase in KYAT3 expression induced by LPS.

[0383] Figure 6F shows the relative fold gene expression of interleukin-6 (IL-6) in brain tissue as measured by qPCR for cohorts from all three groups of endotoxemic pigs: LPS, 20 mg, and 40 mg. CER-001 partially reversed the increase in IL-6 expression induced by LPS.

[0384] This preclinical data indicates that CER-001 treatment reduces kynurenine pathway dysfunction by increasing tryptophan levels, decreasing levels of the kynurenine pathway products quinolinic acid, kynurenic acid, and kynurenine, decreasing the kynurenine / tryptophan ratio, reversing increases in IDO1 expression, and increasing DDC expression (which are expected to result in increased serotonin levels, as well as decreased levels of formylkynurenine and subsequent kynurenine pathway biomarkers). Kynurenine pathway activity is associated with various diseases and conditions, including sepsis-induced cognitive deficits ("brain fog").

[0385] 7.2. Example 2: Randomized Pilot Study Comparing Short-Term CER-001 Infusions at Different Doses to Prevent Sepsis-Induced Acute Kidney Injury Currently, there are no approved treatments for sepsis-associated AKI. Given that the inflammatory response to endotoxemia is a major cause of hemodynamic instability and progression to AKI in patients with sepsis, the primary objective of this study was to investigate whether the use of CER-001 at different doses in combination with standard of care (SOC) treatment is safe and effective, providing a new strategy for treating patients with sepsis, reducing the inflammatory response, and preventing progression to AKI. Without being bound by theory, the anticipated mechanism of action is dual and involves both endotoxin binding by CER-001 and a direct anti-inflammatory effect of CER-001.

[0386] The study reported in this example included 20 patients with Gram-negative sepsis, who were at high risk for acute kidney injury due to high levels of endotoxin activity and impaired function of one or more organ systems. Patients received standard of care treatment, either alone or in combination with one of three dosing regimens of CER-001 (5 patients per group). The study investigated whether the use of CER-001 at different doses in combination with standard of care (SOC) treatment is safe and effective, offering a potential new strategy for treating patients with sepsis, reducing the inflammatory response to endotoxin and preventing progression to AKI according to the Kidney Disease: Improving Global Outcomes (KDIGO) criteria, and the safety and tolerability of the dosing regimens were investigated to select the optimal dose of CER-001.

[0387] One of the metabolic hallmarks of bacterial (as in sepsis) or viral infections (as in SARS-CoV-2) is a strong decrease in circulating lipoproteins, especially high-density lipoproteins (HDL), whose major component is apolipoprotein AI (ApoA-I). As an example, ApoA-I levels were recently described as a biomarker predicting long-term mortality after surgical sepsis.

[0388] One goal of the study was to restore ApoA-I levels using CER-001 to re-establish full functionality of this personalized biomarker and provide benefit in sepsis pathology.

[0389] 7.2.1. Test Protocol Study Population: This was a single-center, randomized, dose-ranging (Phase II) study that included patients with sepsis or urosepsis due to intra-abdominal infection admitted to the intensive care unit (ICU) of the participating centers. The investigators ensured that all patients who met the following inclusion and exclusion criteria were offered enrollment in the study.

[0390] Inclusion Criteria: - Male or non-pregnant female adults aged 18 years or older at the time of enrollment; -SOFA score on admission to SOFA fulfilling sepsis 3 criteria, defined as an acute increase of at least 2 points in the score; endotoxin levels greater than -0.6 (as measured by Endotoxin Activity Assay (EEA™); Spectral Medical) (see Marshall et al., 2004, J Infect Dis. 190(3):527-34); - Signed and dated informed consent from the patient or their legal representative.

[0391] Exclusion criteria: -Patients weighing more than 100 kg; -Alanine transaminase / aspartate transaminase (ALT / AST) greater than 5 times the upper limit of normal; Stage 4 severe chronic kidney disease or requiring dialysis (i.e., ≥30 ml / min / 1.73 m 2 less than estimated glomerular filtration rate (eGFR); -Less than 2.0 x 10^9 white blood cells; - Pregnant or lactating; - received an organ transplant in the past year; - Anticipated transfer to another non-research hospital within 72 hours; - terminal illness, including metastasis or hematologic malignancy, with a life expectancy of less than 30 days (as assessed by the attending physician) or classified as "do not resuscitate"; - Prior history of end-stage chronic organ failure(s); - diagnosed with HIV; - Uncontrollable bleeding within the past 24 hours - Patients who have used an investigational drug or device within 30 days of the first dose of CER-001.

[0392] Number of subjects: 20 subjects were enrolled and randomized (1:1:1:1) into four experimental groups, Groups A-D, as defined below. Baseline characteristics of the subjects are summarized in the table below. [Table 2-1] [Table 2-2]

[0393] The study population had the following baseline clinical and demographic characteristics: [Table 3]

[0394] Duration of the study: The study was completed in 24 weeks (6 months). The enrollment period was approximately 20 weeks (5 months) from the first enrolled subject. The end of the study was the last visit of the last subject.

[0395] Primary Endpoints: The co-primary endpoints of the study were (1) to define the safety and optimal dose of CER-001 in combination with standard of care in patients with persistent sepsis due to Gram-negative bacteria, (2) to determine the onset of AKI according to the KDIGO criteria, and (3) to determine the severity of AKI according to the KDIGO criteria.

[0396] Secondary endpoints: The secondary endpoints were: - Changes in endotoxin and IL-6 levels from baseline to days 3, 6, and 9. Baseline is defined as the last measurement taken before dosing on Day 1. - Change in SOFA score (Vincent et al. 1996, Intensive Care Med, 22:707-710) from baseline to days 3, 6, and 9. - Changes in key inflammatory markers (CRP, D-dimer, ferritin, IL-8, GM-CSF, MCP 1, and TNF-α) from baseline to days 3, 6, and 9. - Changes in AKI biomarkers and the development of AKI according to the KDIGO criteria (Kidney Disease Improving Global Outcomes. KDIGO Clinical Practice Guideline for Acute Kidney Injury. Kidney International Supplements 2012;2:1-138) Mortality at 30 days

[0397] Intervention / Exposure: Twenty patients who met the eligibility criteria and signed and dated informed consent documents approved by the Ethics Committee (EC) were randomized and assigned in a (1:1:1:1) ratio to receive conventional therapy (Group A), low dose CER-001 (Group B), medium dose CER-001 (Group C), or high dose CER-001 (Group D). Conventional therapy was adjusted according to clinical status. All non-experimental treatments were allowed to be administered concurrently during patient participation in this study. Any medication taken by patients other than the study drug specified according to the protocol was considered as concurrent medication and recorded in the study records.

[0398] Each patient was identified by a patient number at screening. Patient numbers were not reused when patients were assigned. The investigators who enrolled the patients did not participate in the randomization and allocation assignment. The randomization list, divided into blocks, was appropriately concealed to prevent attempts to subvert the randomization.

[0399] Treatment group: All patients received conventional therapy. The treatment group received additional therapy with the study drug. In particular, - Group A: Conventional therapy (i.e., antibiotic treatment and hemodynamic support according to the patient's condition). - Group B: Conventional therapy + 5 mg / kg BID for 3 consecutive days, followed by CER-001 at 5 mg / kg BID on day 6. - Group C: Conventional therapy + 10 mg / kg BID for 3 consecutive days followed by CER-001 at 10 mg / kg BID on day 6. - Group D: Conventional therapy + 20 mg / kg BID for 3 consecutive days followed by CER-001 at 20 mg / kg BID on day 6.

[0400] Figure 7 summarizes the study regimen.

[0401] Patients were pretreated with an antihistamine (eg, 5 mg dexchlorpheniramine or 100 mg hydroxyzine) before each CER-001 dose to avoid any potential infusion reactions.

[0402] Statistical Analysis: Comparisons between groups were performed using appropriate statistical tests. Dichotomous variables (baseline characteristics, mortality, and development of AKI) were compared using chi-square or Fisher's exact test, and continuous baseline characteristics were compared by ANOVA or Kruskal-Wallis test, Student's t, or Mann-Whitney U test, as appropriate. Changes in inflammatory markers were compared between groups by ANOVA and presented graphically. The proportion of patients with AKI and mortality were calculated for each group. All analyses were performed using SPSS 12.0 for Windows, and p<0.05 was considered statistically significant.

[0403] Procedures: The following procedures were performed during the screening visit: After randomization, subjects began treatment within 2 weekdays. -Informed consent - Medical History - included recording past and present illnesses and collecting subject demographic data (date of birth, sex, and race). -Physical examination with review of height, weight, BMI and circumference -Vital signs (pulse, blood pressure, and oral, ear, axillary, or core temperature). -Review of inclusion / exclusion criteria. Adverse events were recorded starting from the time informed consent was obtained. - Previous medications were collected from 4 weeks prior to the first dose of study article. All current medications were recorded. Complete blood count (CBC) - included white blood cell count (WBC) with differential, platelet count, red blood cell count (RBC), hemoglobin (Hb), and hematocrit (Hct). -Fasting chemistry panel / electrolytes: including sodium, potassium, chloride, blood urea nitrogen (BUN or urea), serum creatinine, calculated creatinine clearance (CKD-EPI), glucose, calcium, phosphorus, total protein, uric acid, AST, ALT, gamma GT, ALP, total and direct bilirubin, albumin, total cholesterol, HDL, LDL, triglycerides, LDH, CPK, -ABG (to assess respiratory and / or metabolic disorders) -ApoA-I (for pharmacokinetic and pharmacodynamic evaluation) - Coagulation tests - included prothrombin time (PT) (expressed as international normalized ratio [INR]), and partial thromboplastin time (PTT). -Urinary analysis - included assessment of specific gravity, pH, protein / albumin, glucose, ketones, and hemoglobin / blood. Microalbuminuria and proteinuria at -g / 24 hours - Serum or urine pregnancy test (for women of childbearing potential) within 7 days prior to randomization. Pharmacokinetic and pharmacodynamic assessments included ApoA-I and total cholesterol levels. Endotoxin levels were measured using the EAA™ kit. AKI biomarkers (TIMP-2 and IGFBP-7) were measured using the Nephrocheck® kit. Inflammatory markers included CRP, D-dimer, ferritin, IL-6, IL-8, GM-CSF, MCP 1, and TNF-α.

[0404] In addition to the biological samples collected for daily routine clinical laboratory evaluations performed in a central laboratory, biological samples were collected for research purposes and included the following: - Two tubes of 5ml serum -One tube of 3 ml of plasma -30ml of urine

[0405] Using these samples, additional inflammatory cytokines and urinary biomolecules were evaluated to obtain a more comprehensive characterization of enrolled patients, better assess response to treatment, provide more information in follow-up, and, more importantly, discover new potential biomarkers that may be useful for the early diagnosis of sepsis-induced AKI. Analyses were performed by ELISA tests and protein arrays.

[0406] Therapy Visits (Treatment Period): Treatment period was defined as from the start of treatment. Visits were scheduled on days 3, 6, and 9. A final visit was scheduled on day 30. The following procedures were performed during the therapy visits: - Records of adverse events and concomitant medications -Review of appropriate laboratory information -Physical examination - Vital signs (pulse, blood pressure, and oral, ear, axillary, or core temperature) were assessed -Continuously record adverse events and concomitant medications Complete blood count (CBC) - included white blood cell count (WBC) with differential, platelet count, red blood cell count (RBC), hemoglobin (Hb), and hematocrit (Hct). -Fasting chemistry panel / electrolytes: including sodium, potassium, chloride, blood urea nitrogen (BUN or urea), serum creatinine, and calculated creatinine clearance (CKD-EPI), -Glucose, calcium, phosphorus, total protein, uric acid, AST, ALT, □GT, ALP, total and direct bilirubin, albumin, total cholesterol, HDL, LDL, triglycerides, LDH, CPK -ABG (to assess respiratory and / or metabolic disorders) -ApoA-I (for pharmacokinetic and pharmacodynamic evaluation) - Coagulation tests - included prothrombin time (PT) (expressed as international normalized ratio [INR]), and partial thromboplastin time (PTT). -Urinary analysis - included assessment of specific gravity, pH, protein / albumin, glucose, ketones, and hemoglobin / blood. Microalbuminuria and proteinuria at -g / 24 hours - Serum or urine pregnancy test (for women of childbearing potential) within 7 days prior to randomization. Pharmacokinetic and pharmacodynamic assessments included ApoA-I and total cholesterol levels. Endotoxin levels were measured using the EAA™ kit. AKI biomarkers (TIMP-2 and IGFBP-7) were measured using the Nephrocheck® kit. Inflammatory markers included CRP, D-dimer, ferritin, IL-6, IL-8, GM-CSF, MCP 1, and TNF-α.

[0407] In addition to the biological samples collected for daily routine clinical laboratory evaluations performed in a central laboratory, biological samples were collected for research purposes and included the following: - Two tubes of 5ml serum -One tube of 3 ml of plasma -30ml of urine

[0408] Clinical scores included the SOFA score (Table 2) and the KDIGO criteria for AKI assessment and staging (Table 3). The individual components of each score were documented. [Table 4] [Table 5]

[0409] Table 4 provides a summary of the study protocol for this example. [Table 6]

[0410] Safety Assessment: Safety assessment was accomplished utilizing information collected from the following assessments: physical examination (including weight), vital signs (blood pressure, pulse, temperature), CBC with percentages, platelet count, blood chemistry, and fasting lipid profile [including HDL-cholesterol, LDL-cholesterol, and lipoprotein(a)], urea, glucose, 24-hour urine protein measurement, serum creatinine and calculated creatinine clearance (CKD-EPI), and adverse event monitoring. All women of childbearing potential underwent a qualitative serum pregnancy test during pre-study screening / baseline assessment and thereafter as clinically indicated. Patients were monitored for the occurrence of adverse events throughout the study, and adverse events were recorded. Adverse events spontaneously reported by subjects or discovered as a result of general questioning by the investigator or by physical examination were recorded. Duration (start and end dates), severity, cause and relationship to study medication, patient outcome, action taken, and an assessment of whether the event was serious were recorded for each reported adverse event.

[0411] 7.2.2.Results Lipopolysaccharide (LPS) Figures 8A-8F show lipopolysaccharide (LPS) changes for the standard care group (Group A) and experimental groups (Groups B-D). Figure 8A: LPS change from baseline for Group A and Groups B-D. Figure 8B: LPS change from baseline for Groups A, B, C, and D. Figure 8C: LPS change reported as a percentage of peak LPS levels (peak = 100%) for Group A and Groups B-D. The treatment x study day effect for peak was p<0.0005. Figure 8D: LPS change for Group A and Groups B-D, separated by whether the subject was enrolled from the center's ICU or nephrology department. Figure 8E: LPS change from baseline for each subject in Group A and Groups B-D. Figure 8F: LPS change from baseline for each subject in each of Groups A-D. LPS levels were measured by ELISA. Statistically significant differences were assessed by using mixed model ANOVA (ns: p>0.05).

[0412] In general, treatment regimens providing 5 mg / kg, 10 mg / kg, or 20 mg / kg of CER-001 in addition to SOC reduced LPS more than SOC alone. Significant or near-significant results are summarized in the table below. [Table 7]

[0413] Consistent with the animal data and the observed ApoA-I increase, treatment with CER-001 significantly reduced LPS bloodstream concentrations compared to SOC controls (Figure 8E). This observation reinforces the hypothesis of a positive impact of LPS reduction on clinical outcomes. However, other studies have emphasized the need to extend such hypotheses beyond simple LPS removal (Monard et al., 2023, Critical Care 27(1):36; Cavaillon et al., 2020, EMBO Molecular Medicine 12(4)). Indeed, pleiotropic effects, including LPS reduction / inactivation and inhibition of the cytokine storm cascade and / or endothelial dysfunction, could have potent biochemical and clinical implications. Without being bound by theory, it is believed that CER-001 not only reduces LPS levels in animals and humans, but also has direct interactions with the immune system via ApoA-I and provides endothelial protection.

[0414] 7.2.2.2. Endotoxin Activity Assay (EAA) Figures 9A-9D show endotoxin activity assay (EAA) changes for the standard treatment group (Group A) and experimental groups (Groups B-D). Figure 9A: EAA change from baseline for Group A and Groups B-D. Figure 9B: EAA change from baseline for Groups A, B, C, and D. Figure 9C: EAA change reported as a percentage of peak EAA levels (peak = 100%) for Group A and Groups B-D. The treatment x study day effect for peak was p<0.1769. Figure 9D: EAA change for Group A and Groups B-D, separated by whether subjects were enrolled from the center's ICU or nephrology department. Figure 9E: EAA change from baseline for Group A and Groups B-D. EAA was performed at each time point using a commercially available kit (Spectral Medical, Toronto, Canada). Statistically significant differences were assessed using mixed-model ANOVA (ns: p>0.05).

[0415] Overall, treatment regimens providing 10 mg / kg CER-001 in addition to SOC reduced EAAs more than SOC alone. Significant or near-significant results are summarized in the table below. [Table 8]

[0416] TNF-α Figures 10A-10F show the TNF-α change for the standard care group (Group A) and experimental groups (Groups B-D) measured by ELISA. Statistically significant differences were assessed by using a mixed-model ANOVA (ns: p>0.05). Figure 10A: TNF-α change from baseline for Group A and Groups B-D. Figure 10B: TNF-α change from baseline for Groups A, B, C, and D. Figure 10C: TNF-α change reported as a percentage of peak TNF-α levels (peak = 100%) for Group A and Groups B-D. The treatment x study day effect for peak was p<0.0004. Figure 10D: TNF-α change for Group A and Groups B-D, separated by whether the subject was enrolled from the center's ICU or nephrology department. Figure 10E: TNF-α change from baseline for each subject in Group A and Groups B-D. FIG. 10F: TNF-α change from baseline for each subject in each of Groups AD.

[0417] Overall, treatment regimens providing 10 mg / kg CER-001 in addition to SOC reduced TNF-α more than SOC alone.

[0418] 7.2.2.4.MCP-1 Figures 11A-11F show MCP-1 changes for the standard care group (Group A) and experimental groups (Groups B-D) measured by ELISA. Statistically significant differences were assessed by using a mixed-model ANOVA (ns: p>0.05). Figure 11A: MCP-1 change from baseline for Group A and Groups B-D. Figure 11B: MCP-1 change from baseline for Groups A, B, C, and D. Figure 11C: MCP-1 change reported as a percentage of peak MCP-1 levels (peak = 100%) for Group A and Groups B-D. The treatment x study day effect for peak was p<0.0090. Figure 11D: MCP-1 change for Group A and Groups B-D, separated by whether subjects were enrolled from the center's ICU or nephrology department. Figure 11E: MCP-1 change from baseline for Group A and Groups B-D. FIG. 11F: MCP-1 change from baseline for each subject in each of Groups AD.

[0419] Interleukin-6 (IL-6) Figures 12A-12F show IL-6 changes for the standard care group (Group A) and experimental groups (Groups B-D) measured by ELISA. Statistically significant differences were assessed by using a mixed-model ANOVA (ns: p>0.05). Figure 12A: IL-6 change from baseline for Group A and population groups B-D. Figure 12B: IL-6 change from baseline for Groups A, B, C, and D. Figure 12C: IL-6 change reported as a percentage of peak IL-6 levels (peak = 100%) for Group A and population groups B-D. The treatment x study day effect for peak was p<0.0037. Figure 12D: IL-6 change for Group A and population groups B-D, separated by whether the subject was enrolled from the center's ICU or nephrology department. Figure 12E: IL-6 change from baseline for each subject in Group A and population groups B-D. FIG. 12F: IL-6 change from baseline for each subject in each of Groups AD.

[0420] Overall, treatment regimens providing CER-001 in addition to SOC and specific CER-001 at 10 mg / kg reduced IL-6 more than SOC alone. Significant or near-significant results are summarized in the table below. [Table 9]

[0421] Interleukin-8 (IL-8) Figures 13A-13F show IL-8 changes for the standard care group (Group A) and experimental groups (Groups B-D). Figure 13A: IL-8 change from baseline for Group A and population groups B-D measured by ELISA. Statistically significant differences were assessed by using a mixed-model ANOVA (ns: p>0.05). Figure 13B: IL-8 change from baseline for Groups A, B, C, and D. Figure 13C: IL-8 change reported as a percentage of peak IL-8 levels (peak = 100%) for Group A and population groups B-D. The treatment x study day effect for peak was p<0.0001. Figure 13D: IL-8 change for Group A and population groups B-D, separated by whether the subject was enrolled from the center's ICU or nephrology department. Figure 13E: IL-8 change from baseline for each subject in Group A and population groups B-D. FIG. 13F: IL-8 change from baseline for each subject in each of Groups AD.

[0422] Overall, treatment regimens providing CER-001 in addition to SOC reduced IL-8 more than SOC alone. Significant or near-significant results are summarized in the table below. [Table 10]

[0423] Interleukin-10 (IL-10) Figures 14A-14D show IL-10 changes for the standard care group (Group A) and experimental groups (Groups B-D) measured by ELISA. Statistically significant differences were assessed by using a mixed-model ANOVA (ns: p>0.05). Figure 14A: IL-10 change from baseline for Group A and population Groups B-D. Figure 14B: IL-10 change from baseline for Groups A, B, C, and D. Figure 14C: IL-10 change reported as a percentage of peak IL-10 levels (peak = 100%) for Group A and population Groups B-D. The treatment x study day effect for peak was p<0.3780. Figure 14D: IL-10 change for Group A and population Groups B-D, separated by whether subjects were enrolled from the center's ICU or nephrology department.

[0424] 7.2.2.8.TREM-1 Figures 15A-15F show TREM-1 changes for the standard care group (Group A) and experimental groups (Groups B-D) measured by ELISA. Statistically significant differences were assessed by using a mixed-model ANOVA (ns: p>0.05). Figure 15A: TREM-1 change from baseline for Group A and population groups B-D. Figure 15B: TREM-1 change from baseline for Groups A, B, C, and D. Figure 15C: TREM-1 change reported as a percentage of peak TREM-1 levels (peak = 100%) for Group A and population groups B-D. The treatment x study day effect for peak was p<0.0003. Figure 15D: TREM-1 change for Group A and population groups B-D, separated by whether the subject was enrolled from the center's ICU or nephrology department. Figure 15E: TREM-1 change from baseline for each subject in Group A and population groups B-D. FIG. 15F: TREM-1 change from baseline for each subject in each of Groups AD.

[0425] Overall, treatment regimens providing CER-001 in addition to SOC reduced TREM-1 more than SOC alone. Significant or near-significant results are summarized in the table below. [Table 11]

[0426] VCAM and ICAM Figures 16A-16F show VCAM changes for the standard care group (Group A) and experimental groups (Groups B-D) measured by ELISA. Statistically significant differences were assessed by using a mixed-model ANOVA (ns: p>0.05). Figure 16A: VCAM change from baseline for Group A and population Groups B-D. Figure 16B: VCAM change from baseline for Groups A, B, C, and D. Figure 16C: VCAM change reported as a percentage of peak VCAM levels (peak = 100%) for Group A and population Groups B-D. The treatment x study day effect for peak was p<0.0001. Figure 16D: VCAM change for Group A and population Groups B-D, separated by whether the subject was enrolled from the center's ICU or nephrology department. Figure 16E: VCAM change from baseline for each subject in Group A and population Groups B-D. Figure 16F: VCAM change from baseline for each subject in each of Groups A-D.

[0427] Overall, treatment regimens providing CER-001 in addition to SOC reduced VCAM more than SOC alone. Significant or near-significant results are summarized in the table below. [Table 12]

[0428] Figures 17A-17F show ICAM changes for the standard care group (Group A) and experimental groups (Groups B-D) measured by ELISA. Statistically significant differences were assessed by using a mixed-model ANOVA (ns: p>0.05). Figure 17A: ICAM change from baseline for Group A and population Groups B-D. Figure 17B: ICAM change from baseline for Groups A, B, C, and D. Figure 17C: ICAM change reported as a percentage of peak ICAM levels (peak = 100%) for Group A and population Groups B-D. The treatment x study day effect for peak was p<0.0001. Figure 17D: ICAM change for Group A and population Groups B-D, separated by whether the subject was enrolled from the center's ICU or nephrology department. Figure 17E: ICAM change from baseline for each subject in Group A and population Groups B-D. Figure 17F: ICAM change from baseline for each subject in each of Groups A-D.

[0429] Overall, treatment regimens providing CER-001 in addition to SOC reduced ICAM more than SOC alone. Significant or near-significant results are summarized in the table below. [Table 13]

[0430] Ferritin Figures 18A-18D show ferritin changes for the standard care group (Group A) and experimental groups (Groups B-D). Figure 18A: Ferritin change from baseline for Group A and population Groups B-D. Figure 18B: Ferritin change from baseline for Groups A, B, C, and D. Figure 18C: Ferritin change reported as a percentage of peak ferritin level (peak = 100%) for Group A and population Groups B-D. The treatment x study day effect for peak was p<0.0962. Figure 18D: Ferritin change for Group A and population Groups B-D, separated by whether subjects were enrolled from the center's ICU or nephrology department.

[0431] 7.2.2.11.White blood cells Figures 19A-19D show white blood cell count changes for the standard care group (Group A) and experimental groups (Groups B-D). Figure 19A: White blood cell count change from baseline for Group A and population Groups B-D. Figure 19B: White blood cell count change from baseline for Groups A, B, C, and D. Figure 19C: White blood cell count change reported as a percentage of peak white blood cell count (peak = 100%) for Group A and population Groups B-D. The treatment x study day effect for peak was p = 0.5492. Figure 19D: White blood cell count change for Group A and population Groups B-D, separated by whether subjects were enrolled from the center's ICU or nephrology department.

[0432] C-reactive protein Figures 20A-20F show CRP changes for the standard care group (Group A) and experimental groups (Groups B-D). Statistically significant differences were assessed by using a mixed-model ANOVA (ns: p>0.05). Figure 20A: CRP change from baseline for Group A and population Groups B-D. Figure 20B: CRP change from baseline for Groups A, B, C, and D. Figure 20C: CRP change reported as a percentage of peak CRP level (peak = 100%) for Group A and population Groups B-D. The treatment x study day effect for peak was p<0.6446. Figure 20D: CRP change for Group A and population Groups B-D, separated by whether the subject was enrolled from the center's ICU or nephrology department. Figure 20E: CRP change from baseline for each subject in Group A and population Groups B-D. Figure 20F: CRP change from baseline for each subject in each of Groups A-D.

[0433] 7.2.2.13.KIM-1 Figures 21A-21D show KIM-1 changes for the standard care group (Group A) and experimental groups (Groups B-D). Figure 21A: KIM-1 change from baseline for Group A and population Groups B-D. Figure 21B: KIM-1 change from baseline for Groups A, B, C, and D. Figure 21C: KIM-1 change reported as a percentage of peak KIM-1 levels (peak = 100%) for Group A and population Groups B-D. The treatment x study day effect for peak was p<0.0001. Figure 21D: KIM-1 change for Group A and population Groups B-D, separated by whether subjects were enrolled from the center's ICU or nephrology department.

[0434] Serum Albumin Figures 22A-D show serum albumin changes for the standard care group (Group A) and experimental groups (Groups B-D). Figure 22A: Serum albumin change from baseline for Group A and population Groups B-D. Figure 22B: Serum albumin change from baseline for Groups A, B, C, and D. Figure 22C: Serum albumin change reported as a percentage of peak serum albumin level (peak = 100%) for Group A and population Groups B-D. The treatment x study day effect for peak was p = 0.1595. Figure 22D: Serum albumin change for Group A and population Groups B-D, separated by whether the subject was enrolled from the center's ICU or nephrology department. Figure 22E: Serum albumin change from baseline for each subject in Group A and population Groups B-D.

[0435] Overall, treatment regimens providing CER-001 in addition to SOC increased serum albumin more than SOC alone.

[0436] The significant or near-significant results are summarized in the table below. [Table 14]

[0437] Serum creatinine Figures 23A-F show serum creatinine changes for the standard care group (Group A) and experimental groups (Groups B-D). Figure 23A: Serum creatinine change from baseline for Group A and population Groups B-D. Figure 23B: Serum creatinine change from baseline for Groups A, B, C, and D. Figure 23C: Serum creatinine change reported as a percentage of peak serum creatinine level (peak = 100%) for Group A and population Groups B-D. The treatment x study day effect for peak was p = 0.1630. Figure 23D: Serum creatinine change for Group A and population Groups B-D, separated by whether the subject was enrolled from the center's ICU or nephrology department. Figure 23E: Area under the curve (AUC) (mean ± SEM) for serum creatinine for Group A and population Groups B-D for all subjects and for subject populations from the ICU and nephrology intake pathways. Figure 23F: AUC (95% confidence interval) for serum creatinine for Group A and aggregate Groups BD, and subject populations from the ICU and nephrology uptake pathways.

[0438] Estimated glomerular filtration rate (eGFR) Figures 24A-B show eGFR changes for all subjects in the standard care group (Group A) and experimental groups (Groups B-D). Estimated GFR was determined by CKD-EPI. Figure 24A: eGFR change from baseline for Group A and population Groups B-D. Figure 24B: eGFR change from baseline for Groups A, B, C, and D. Figure 24E shows eGFR change, reported as a percentage of peak level (peak = 100%), for Group A and population Groups B-D for all subjects. The treatment x study day effect for peak was p = 0.5666.

[0439] Figures 24C-D show eGFR changes for subjects with AKI who entered the study. Figure 24C: eGFR change from baseline for Group A and population groups B-D. Figure 24D: eGFR change for Groups A, B, C, and D. Figure 24F shows eGFR change, reported as a percentage of peak level (peak = 100%), for Group A and population groups B-D for subjects with AKI who entered the study. The treatment x study day effect for peak was p = 0.2406.

[0440] The significant or near-significant results from Figures 24C-24D are summarized in the table below. [Table 15]

[0441] 7.2.2.17.P / F ratio FIG. 25 shows the change in P / F ratio for all subjects in the standard care group (Group A) and population groups BD.

[0442] 7.2.2.18. Number of days in ICU FIG. 26 shows the survival percentage for all subjects after days in ICU for the standard of care group (Group A, "SOC") and collective groups BD ("CER-001").

[0443] 7.2.2.19.30 days alive FIG. 27A shows the 30-day survival rates for all subjects for the standard of care group (Group A, "SOC") and collective groups BD ("CER-001").

[0444] FIG. 27B shows the 30-day survival rate for all subjects enrolled in the study from the center's ICU for the standard of care group (Group A, "SOC") and aggregate groups BD ("CER-001").

[0445] AKI Staging Figure 28A shows the evolution of AKI as assessed by the KDIGO staging criteria for all subjects in the standard of care group (Group A, "SOC"). AKI stages: 0, serum creatinine less than 1.5 times baseline or increased by less than 0.3 mg / dl within 48 hours, and a urine volume greater than 0.5 ml / kg / hour for 6-12 hours; 1, serum creatinine 1.5-1.9 times baseline or increased by more than 0.3 mg / dl, or a urine volume less than 0.5 ml / kg / hour for 6-12 hours; 2, serum creatinine 2.0-2.9 times baseline, or a urine volume less than 0.5 ml / kg / hour for more than 12 hours; 3, serum creatinine 3.0 times baseline or greater than 4.0 mg / dl, or a urine volume less than 0.3 ml / kg / hour for more than 24 hours or approximately zero for more than 12 hours. On day 6, approximately 40% of SOC subjects had AKI 0 (minimum severity), and the remainder had AKI 2-3 (more severe to maximal severity).

[0446] Figure 28B shows the evolution of AKI as assessed by the KDIGO staging criteria for all subjects in population groups B through D ("CER-001"). At day 6, approximately 60% of CER-001 subjects had AKI 0 and 20% had AKI 3.

[0447] 7.2.2.21. Days on mechanical ventilation and vasopressors FIG. 29 shows the number of days on mechanical ventilation for all subjects who participated in the study while in the center's ICU for the standard of care group (Group A, SOC) and cluster groups B-D (CER-001). CER-001 reduced the number of days on mechanical ventilation for 5 / 7 subjects relative to SOC.

[0448] FIG. 30 shows the number of days on vasopressors for all subjects who participated in the study while in the center's ICU for the standard of care group (Group A, SOC) and aggregate groups BD (CER-001).

[0449] 7.2.2.22. Number of days on dialysis Figure 31A shows the number of days on dialysis for all subjects who participated in the study while in the center's ICU for the standard care group (Group A) and population groups B-D. Figure 31B shows this result for all subjects. Both intermittent and continuous modalities were considered. A reduction in the number of days on dialysis indicates improved kidney function.

[0450] 7.2.2.23. Days of survival without organ support, days to ICU discharge, and hemodynamic changes Figure 32 shows the organ support-free survival in days for all subjects who participated in the study while in the center's ICU for the standard of care group (SOC) and cohort groups B-D (CER-001). The use of any vasopressors, mechanical ventilation, and / or renal support was considered organ support.

[0451] Figure 65 shows the number of days to ICU discharge for all subjects enrolled in the study from the center's ICU for the standard of care group (SOC) and cluster groups BD (CER-001).

[0452] Figures 33A and 33B show the change in daily mean arterial pressure (MAP) for all subjects who participated in the study while in the center's ICU for the standard of care group (SOC) and cluster groups B through D (CER-001). A reduction in MAP is generally desirable for ICU subjects.

[0453] Figure 34 shows the change in average daily heart rate (HR) for all subjects who participated in the study while in the center's ICU for the standard of care group (SOC) and cohort groups B-D (CER-001). A reduction in MAP is generally desirable for ICU subjects.

[0454] Figure 35 shows the change in the average daily P / F ratio for all subjects who participated in the study while in the center's ICU for the standard of care group (SOC) and cohort groups B through D (CER-001). An increase in the P / F ratio is generally desirable for ICU subjects.

[0455] 7.2.2.24. Serum ApoA-I FIG. 56A shows the mean ApoA-I levels for the control and population study groups in the clinical study of Example 2.

[0456] Figure 56 shows the mean ApoA-I levels for the control group and the combined study group; Figure 56B shows the mean ApoA-I levels for the control group and each study group; Figure 56C shows the ApoA-I change for each subject in the standard of care (SOC) and experimental (CER-001) groups; and Figure 56D shows the change from baseline in ApoA-I levels for each subject divided by study group, as measured by ELISA. Serum ApoA-I levels increased rapidly in patients receiving CER-001 during the first three days of treatment, with a delayed increase in the SOC group. Statistically significant differences were assessed using mixed-model ANOVA (ns: p>0.05).

[0457] Kynurenine pathway markers FIG. 65 shows the change in serum quinolinic acid (QA) levels from baseline (Day 1) for subjects in the standard of care (SOC) group (Group A) and population groups BD in the clinical study of Example 2.

[0458] FIG. 67 shows the change in serum kynurenine / tryptophan ratio (Kyn / Trp) levels from baseline (Day 1) for subjects in the standard of care (SOC) group (Group A) and population groups B through D in the clinical study of Example 2.

[0459] FIG. 68 shows the change in serum serotonin levels from baseline (Day 1) for subjects in the standard of care (SOC) group (Group A) and population groups BD in the clinical study of Example 2.

[0460] ADDITIONAL RESULTS Across all subjects, liver enzymes changed anywhere from about 0.2 to about 4 times baseline levels after 9 days (Figures 57A, 57B). Subjects receiving CER-001 had similar changes in liver enzyme levels as those receiving standard of care alone.

[0461] Adverse Events: Of the 20 subjects, 9 experienced adverse events, of which 7 were definitely not related to the study article (i.e., the adverse event could be explained entirely by the subject's clinical condition or other medications / therapy) and 2 were probably not related to the study article (i.e., the adverse event could most likely be explained by the subject's clinical condition or other medications / therapy, and not by the study article).

[0462] 7.2.3. Conclusion Overall, co-administration of CER-001 with standard of care resulted in improved clearance of endotoxin (as indicated by LPS and EAA data), modulation of CRS (most evident for IL-6, IL-8, and TREM-1), and endothelial protection (as indicated by VCAM and ICAM data) without increasing inflammation or negatively affecting AKI biomarkers or hemodynamics.

[0463] Notably, the trends of endothelial dysfunction markers VCAM and ICAM showed different trends from ApoA-I among study participants: VCAM and ICAM levels increased in patients in the SOC group, whereas VCAM and ICAM significantly decreased with CER-001 treatment (Figures 16E and 17E), suggesting increased vascular protection due to these mediators contributing to the enhancement of irreversible mechanisms of tubular apoptosis and renal injury. These effects are consistent with the in vitro observation of eNOS production by endothelial cells inhibited by CER-001, discussed in Example 4, thus confirming the positive effect of the ApoA-I complex on vascular permeability, a hallmark of acute and chronic inflammation.

[0464] Recently, the anti-inflammatory potential of CER-001 was highlighted in severely ill COVID-19 patients in the ICU, where evaluation of serum amyloid A-1, inflammatory markers, and cytokines primarily showed significant reductions during CER-001 infusion (Begue, et al., 2021, Sci Rep 11, 2291). Similarly, the data in this example showed that CER-001 treatment induced significant reductions in serum levels of MCP-1, TNF-α, IL-6, and IL-8 in treated patients compared with the SOC group (Figures 11E, 10E, 12E, and 13E), suggesting the immunomodulatory and anti-inflammatory effects of CER-001 treatment and its ability to modulate cytokine storm. Consistent with cytokine storm modulation, the treatment group showed a more pronounced reduction in C-reactive protein (CRP) over the first 9 days compared with the SOC group (Figure 20E).

[0465] Soluble triggering receptor-1 (sTREM-1), expressed in myeloid cells, has been suggested as a strong predictor of poor prognosis and survival in sepsis patients. Persistently elevated sTREM-1 levels during the first days after ICU admission are associated with mortality in human septic shock (Jolly, et al., 2021, Cell Mol Immunol 18, 2054-2056). The results of this example suggest that sTREM-1 rapidly decreases within the first 3 days of CER-001 treatment and remains low and stable for at least 30 days (Figure 15E). More importantly, this decrease is accompanied by a resolution of clinical signs and symptoms. Collectively, these observations confirm the potential anti-inflammatory effects of ApoA-I complexes, independent of LPS removal. Understanding that HDL and ApoA-I can interact directly with monocytes / macrophages either through receptor interactions, e.g., SR-BI, or transporters, e.g., ABCA-1 and / or ABCG-1, or by blocking contact-mediated activation of monocytes / macrophages by T lymphocytes, as evidenced by decreased TNF-α (Figure 10E), one can hypothesize that the direct cellular effect of HDL, and therefore CER-001, on such cells results in a general decrease in cytokine production by an as yet unknown mechanism.

[0466] The potential impact of cytokine cascades and endothelial dysfunction on clinical outcomes was evaluated given the observed potent effects of CER-001.

[0467] The effect on organ dysfunction was analyzed with the hypothesis that the immunomodulatory effects of CER-001 treatment could limit renal dysfunction. To this end, renal function among study participants was analyzed, and AKI onset and severity were classified according to the KDIGO criteria based on both creatinine and urine output criteria. Overall, a lower risk of AKI onset and / or progression to moderate-to-severe AKI (AKI stages 2-3) was observed (26.6%) among CER-001-treated patients up to day 6 compared with the SOC group, in which approximately 60% of patients presented with such a condition (Figures 28A-B). These results were consistent with the clinical and histological data reported in animal models, confirming the protective effect of the treatment.

[0468] In addition, the effects of the study drug on liver function were analyzed. As shown in Figures 57A-57B, no significant changes in liver enzymes (AST, ALT) were observed among CER-001 subjects, and only two patients in the treatment group exhibited slight increases in AST and ALT, without clinically significant increases. Conversely, an increase in albumin levels was observed in the CER-001 treatment group compared with the SOC group (Figure 22E). Overall, these results support the safety of CER-001 and the possibility that the early and sustained effects of treatment on inflammatory conditions may improve liver function and increase albumin production.

[0469] The effects of the study drugs on serum levels of kynurenine pathway markers were also investigated. As shown in Figure 66, subjects receiving standard treatment had a slight increase in serum quinolinic acid (QA) levels over the 30-day study period. In contrast, CER-001 reduced QA levels from baseline within 3 days. The effect was substantially sustained until the end of the study period.

[0470] As shown in Figure 67, a similar effect was observed in serum kynurenine / tryptophan ratio (Kyn / Trp). The SOC group had essentially unchanged Kyn / Trp from baseline (Day 1) to the end of the study period. The CER-001 group had an approximately 40% reduction in Kyn / Trp by Day 30.

[0471] Figure 68 shows that serum serotonin levels remained near baseline throughout the study period for subjects in the CER-001 group. The SOC group experienced a significant drop in serum serotonin of approximately 40 ng / mL on day 6, which did not return to baseline levels by the end of the study period.

[0472] We focused our analysis on the subset of critically ill patients enrolled in the ICU to analyze key clinical outcomes. Although the small sample size (7 treated patients, 2 SOC subjects) limits statistical evaluation, a reduced ICU length of stay was observed among patients in the treatment group (mean ICU stay of 23.2 vs. 29 days) (Figure 65). In addition, mean daily arterial pressure (MAP) during the study period improved after the second day of treatment compared with SOC subjects (mean days of 6.5 vs. 8 in the SOC group), who had an overall lower number of days on vasopressors (Figure 33B, Figure 30). Similarly, the number of days on mechanical ventilation (mean days of 16.7 vs. 26.5) was lower among CER-001-treated patients (Figure 29). Both SOC subjects required dialysis during the study period; only three of the seven CER-001-treated patients did so (Figure 31).

[0473] Finally, the need for any form of organ support (a composite endpoint including mechanical ventilation, dialysis, and / or vasopressor use) was lower in treated subjects (mean days of survival without organ support: 5.8 vs. 2) (Figure 32). Overall, these results suggested a more rapid improvement in clinical status among patients receiving CER-001 treatment.

[0474] In particular, the results demonstrate the following: - Direct and significant effect of CER-001 on endotoxin removal and the resulting reduction in the inflammatory cascade or "cytokine storm" - Significant protective effect of CER-001 on endothelial function Trends toward fewer ICU days, lower need for organ support, and improved 30-day survival for treated patients - Reinforces CER-001's well-established safety profile

[0475] The results also demonstrate that CER-001 had effects on three kynurenine pathway biomarkers, effects consistent with a beneficial effect on cognitive impairment or "brain fog."

[0476] 7.3. Example 3: Lipid-binding protein molecular therapy in a porcine model of LPS-induced acute kidney injury This example describes additional materials and methods, results, and analysis of the study of Example 1.

[0477] 7.3.1 Materials and Methods Animal models Animal studies were carried out in domestic pigs (Sus scrofa domesticus) after approval by the ethical committee of the relevant government authority. The number of animals was selected by calculating the number of subjects required for proper analysis using the calculator program Anastat (www.anastats.fr / ). Six pigs per group were evaluated in relation to data obtained in previous published studies of porcine models of acute kidney injury (Castellano et al., 2014, Crit Care 18, 520; Stasi et al., 2021, Front Immunol 12, 605212; Castellano et al., 2019, Int J Mol Sci 20; Sallustio et al., 2019, FASEB J 33, 10753-10766; Curci et al., 2014, Nephrol Dial Transplant 29, 799-808; Castellano et al., 2016, Am J Transplantation 16, 325-333). Statistical analysis was performed at a significance level of p = 0.05. Briefly, endotoxemia was induced by intravenous infusion of saline solution containing 300 μg / kg of LPS (lipopolysaccharide membrane of Escherichia coli). If left untreated, pigs progressed to LPS-induced AKI as previously described (Sallustio, et al., 2019, FASEB J 33, 10753-10766). The goal of the study was to determine whether treatment with CER-001 could prevent the development of AKI. Animals were randomized into three groups: LPS (endotoxemic pigs, n=6), CER20 (endotoxemic pigs treated with a single dose of 20 mg / kg CER-001 from Abionyx Pharma, Toulouse, France, n=6), and CER20x2 (endotoxemic pigs treated with two doses of 20 mg / kg CER-001 at time 0 and 3 hours later, n=6).

[0478] A few minutes after the start of the LPS infusion, the CER20 group was treated with CER-001 infusion via an isolated venous access. The CER20x2 group was treated by administering two doses of CER-001 (20 mg / kg) via the previously isolated venous access. The first dose was administered a few minutes after the start of the LPS infusion (T0), and the second dose was administered 3 hours after the start of the LPS infusion (T3 / T0 bis). For all CER-001 dosing, the drug product was thawed and then diluted with normal saline to a volume of 250 mL containing 20 mg / kg of CER-001, individualized for each animal based on body weight, and administered using an infusion pump at a fixed rate of 250 mL / hour over a 1-hour period. The LPS group received 250 mL of normal saline at the same infusion rate.

[0479] The dose of CER-001 is defined as the concentration of human ApoA-I present in the dosing solution. Surviving animals were sacrificed approximately 24 hours after LPS / saline injection with an overdose of IV propofol, followed immediately by a 10 ml IV bolus of a supersaturated solution of potassium chloride (2 mEq / ml, Galenica Senese, srl, Italy).

[0480] Domestic pigs (Sus scrofa domesticus) were injected with 300 μg / kg LPS and either no (control group) or 20 mg / kg CER-001 (CER20 group), followed by a second injection of 20 mg / kg CER-001 at 3 hours for half of the group (CER20x2 group).

[0481] 7.3.1.2. Sample collection At the time of sacrifice or early death, kidneys and livers were collected from all animals and processed using standard procedures as previously described (Sallustio, et al., 2019, FASEB J 33, 10753-10766). Urine samples were collected from all animals via catheter, and urine volume was recorded hourly. Pig serum was collected at baseline (T0; before LPS infusion) and at intermediate time points up to 24 hours via an indwelling arterial catheter. Bile samples were collected from all animals at sacrifice. LPS was extracted from bile samples using a phenol-water extraction method (Harada, et al., 2003, Lab Invest 83, 1657-1667) with an LPS extraction kit (Intron Biotechnology, Kyungki-Do, Korea) according to the manufacturer's instructions.

[0482] Total protein extraction was performed from all bile samples (Ciordia, et al., 2021, J Proteomics 230, 103984; Ciordia, et al., 2022, Methods Mol Biol 2420, 1-10). 1000 μl of bile from each sample was centrifuged at 9000 × g for 3 minutes at 4 °C, and the supernatant containing soluble proteins was used for assessment of ApoA-I levels.

[0483] Assessment of LPS and ApoA-I levels The Apo-A1 content of serum and bile samples was measured by ELISA assay (R&D Systems, Minneapolis, MN, USA), as was LPS (R&D Systems, Minneapolis, MN, USA).

[0484] 7.3.1.4. Evaluation of Pro-inflammatory Cytokines and Markers of Endothelial Dysfunction Serum IL-6 and TNF-α levels were measured by ELISA (R&D Systems, Minneapolis, MN, USA), as were s-VCAM, s-ICAM, and MCP-1 (MyBioSource, San Diego, CA, USA).

[0485] 7.3.1.5. Assessment of Classical, Lectin, and Alternative Complement Pathway Activity Complement function in pig serum was assessed using an ELISA (WIESLAB® Complement System Screen COMPL 300, Euro-Diagnostica) as previously described (Castellano, et al., 2010, Am J Pathol 176, 1648-1659).

[0486] 7.3.1.6. Kidney and Liver Function Measurements Serum / urine creatinine, serum / urine kidney injury molecule-1 (KIM-1), and serum / urine cystatin C measurements were performed using commercially available ELISA kits (MyBioSource, San Diego, USA) according to the manufacturer's instructions. Liver function was assessed by serum measurement of the ALT enzyme using a commercially available ELISA (MyBioSource, San Diego, USA).

[0487] 7.3.1.7. Histological Analysis of Kidney and Liver Tissue Kidney and liver tissues were processed for histological staining [hematoxylin and eosin (HE) (Millipore Sigma)]. Digital slides were acquired and analyzed using an AperioScanScope CS2 device (Aperio, Vista, CA, USA) as previously described (Stasi, et al., 2021, Front Immunol 12, 605212; Sallustio, et al., 2019, FASEB J 33, 10753-10766). HE staining was performed to evaluate histological injury in both the kidney and liver. Tubular and glomerular damage were scored semiquantitatively by two blinded observers. The score index for each animal was expressed as the mean of all scores obtained. Both tubular and glomerular pathology scores for each group were expressed as the mean ± SEM. Liver injury was defined as the amount of hepatic lobule destruction, inflammatory cell infiltration, hemorrhage, and hepatocyte necrosis (Baranova, et al., 2016, J Immunol 196, 3135-3147). A score of 1 to 4 was assessed using criteria from a previously published study (ibid.). Pathology scores for each group were expressed as mean ± SEM.

[0488] Western Blot Analysis Liver tissue was homogenized and treated with RIPA lysis buffer (1 mM PMSF, 5 mM EDTA, 1 mM sodium orthovanadate, 150 mM sodium chloride, 8 μg / mL leupeptin, 1.5% Nonidet P-40, and 20 mM Tris-HCl, pH 7.4) containing phosphatase and protease inhibitors. Samples (30 μg of protein) were separated in a 4-15% polyacrylamide gel and then transferred to a PVDF membrane (0.2 mM) using a Trans-Blot Turbo (BioRad, Hercules, CA, USA). Nonspecific binding sites in the blot were blocked by incubation in 5% BSA for 1 hour, and the membrane was then incubated with primary antibodies overnight and secondary antibodies for 1 hour. Immune complexes were detected using the ECL chemiluminescence system (Amersham Pharmacia, Little Chalfont, UK) according to the manufacturer's instructions. The primary antibodies used were anti-LPS (Abcam) and anti-β-actin (1:20,000; Sigma). The secondary antibodies used were HRP-conjugated anti-rabbit (Abcam) and anti-mouse (Abcam). Chemiluminescent blots were acquired using Chemidoc and analyzed using Image J software. Protein expression levels were normalized to those of β-actin.

[0489] 7.3.1.9.Statistical analysis Survival data were analyzed using the log-rank test for trend, and values ​​were censored for surviving animals using the time of sacrifice. LPS, cytokines (TNF-α, MCP-1, IL-6), endothelial markers (VCAM, ICAM), complement (classical, alternative, and lectin pathways), serum (sCR, sKIM-1, sCystatin-C) and urinary (uKIM-1, uCystatin-C) kidney biomarkers, and ALT were analyzed using two-way repeated measures ANOVA corrected for multiple comparisons of pairwise treatment group differences using Tukey's method. For these analyses, data were converted to change from baseline, and the last observation was carried forward as needed to handle any missing values. Urine volume, tubular injury score, glomerular injury score, and liver injury score were analyzed using one-way ANOVA corrected for multiple comparisons of pairwise treatment group differences using Tukey's method. For ANOVA analyses, pairwise treatments were also tested without correction for multiple comparisons. Generally, both the corrected and uncorrected tests agreed on statistically significant findings. The table below shows significant findings from both methods and highlights any differences between Tukey's corrected and uncorrected Fisher's LSD. [Table 16-1] [Table 16-2] [Table 16-3] [Table 16-4] [Table 16-5] JPEG2026504047000023.jpg181156 [Table 16-6] [Table 16-7]

[0490] For in vitro analysis, data are presented as mean ± standard deviation (SD) and compared by Student's t test.

[0491] All analyses were performed by using GraphPad Prism 9.2.0 (GraphPad software, Inc., San Diego, CA, USA).

[0492] 7.3.2.Results Groups of 6.8±0.7-month-old pigs were injected with the indicated doses of LPS and CER-001, and mortality was recorded for 24 hours (n=6 per group). Untreated pigs were highly susceptible to LPS challenge and typically succumbed before completion of the study protocol, with a survival rate of approximately 16.7%. CER-001 treatment increased the median survival of endotoxemic pigs by 50% and 66.7% in the CER20 and CER20x2 groups, respectively (Figure 36). Significant statistical treatment trends were observed in the three groups by log-rank (p=0.02 log-rank trend test).

[0493] Serum levels of VCAM (Figure 37), ICAM (Figure 38), TNF-α (Figure 39), MCP-1 (Figure 40), and IL-6 (Figure 41) were measured by ELISA assay (n=6 independent samples per time point and group). Gray bands indicate two injections (T0-T1 and T3-T4) of saline or CER-001 at a flow rate of 250 ml / h. Results are presented as mean ± SEM. Significant differences were assessed using two-way repeated measures ANOVA with Tukey's correction for multiple comparisons (ns: p>0.05, *p<0.05 **p<0.005, **p<0.0005 vs. LPS group; §p<0.05, §§p<0.0005 vs. CER20 group).

[0494] Systemic complement activation was measured for the classical pathway (Figure 42), alternative pathway (Figure 43), and lectin pathway (Figure 44) by Wieslab assay (n=6 independent samples per time point and group). In each graph, gray bands represent two infusions (0-1 hour and 3-4 hours) of saline or CER-001 at a flow rate of 250 ml / h. Results are presented as mean ± SEM. Significant differences were assessed using two-way repeated measures ANOVA with Tukey's correction for multiple comparisons (ns: p>0.05, *p<0.05 **p<0.005, **p<0.0005 vs. LPS group; §p<0.05, §§p<0.0005 vs. CER20 group).

[0495] As a potent stimulator of the innate immune system, circulating LPS has been primarily described as a rapid inducer of chemokine and cytokine secretion (Tucureanu, et al., 2018, Int J Nanomedicine 13, 63-76). Our results are consistent with this description of a representative set of these pro-inflammatory molecules. TNF-α, typically released early in the inflammatory cascade by monocytes / macrophages, rapidly increased in endotoxemic animals, followed later by increases in MCP-1 and IL-6 (LPS group, Figures 39-41). This increase was significantly impaired by CER-001 (T24, CER20 vs. LPS group: TNF-α, p = 0.0004), with a stronger effect after two doses of CER-001 (T24, CER20 × 2 vs. LPS group: TNF-α, p < 0.0001; MCP-1, p = 0.0009; IL-6, p = 0.0086). As expected, such potent stimulation of the immune system also induced a significant increase in the systemic complement activation cascade, and this increase was significantly inhibited by CER-001 treatment, regardless of the intrinsic activation pathway, i.e., classical, alternative, and lectin pathways, even when the latter was less stimulated by LPS injection.

[0496] Endothelial dysfunction is a key pathological feature in patients with sepsis, and is also prominent in pig models of LPS-induced sepsis (Castellano, et al., 2014, Crit Care 18, 520; Stasi, A. et al., 2021, Front Immunol 12, 605212). Several aspects of sepsis contribute to endothelial dysfunction, such as hemodynamic instability, direct interaction with bacterial components, release of pro-inflammatory cytokines by pathogen-activated immune cells, and procoagulant mediators (Boisrame-Helms, et al., 2013, Curr Vasc Pharmacol 11, 150-160). The subsequent activation of endothelial cells induces the upregulation and expression of different adhesion molecules, such as ICAM and VCAM, which enhance leukocyte migration and transmigration, amplifying innate and adaptive immune responses (de Pablo, R. et al., 2013, Eur J Intern Med 24, 132-138). CER-001 infusion ameliorated systemic endothelial dysfunction by reducing VCAM (Figure 37) and ICAM (Figure 38) serum levels in both treatment groups, with the increased effect of the two ApoA-I complex dose groups highlighted at T6.

[0497] In the continuum of systemic deterioration, liver dysfunction is a severe manifestation in the course of sepsis, primarily caused by alterations to hepatocytes and / or direct and indirect invasion (Yan, et al., 2014, Int Rev Immunol 33, 498-510). Early signs of liver dysfunction were observed in endotoxemic pigs, including increased ALT levels and histological changes (as demonstrated by H&E staining of liver tissue in the LPS and treatment groups (n = 6 independent pigs per group)), such as microvacuolation and infiltrating inflammatory cells (evident in the LPS group, Figure 45A). Scale bar: 80 μm. Liver injury was defined as the amount of destruction of hepatic lobules (black arrows) and infiltration of inflammatory cells (black dashed arrows). Hepatocytes appeared enlarged and hepatic cords were disorganized.

[0498] Liver injury was resolved by CER-001 infusion, as measured by a moderate increase in ALT (Figure 45E) and a statistically significant decrease in liver histological score (Figure 45D). Histological scores were calculated from five randomly selected fields per sample (n = 6 independent pigs per group) shown in Figures 45A-C and scored from 1 to 4 according to the % area of ​​involvement per high-power field, highlighting the protection that CER-001 provides to liver tissue. In Figure 45E, gray bands indicate two infusions (0-1 h and 3-4 h) of saline or CER-001 at a flow rate of 250 ml / h.

[0499] Overproduction of proinflammatory cytokines is associated with sepsis progression, severity, and the development of acute kidney injury (Wang, et al., 2008, Am J Emergency Med 26, 711-715). Renal injury in a pig model was assessed by a time-dependent increase in serum creatinine (Figure 47), and there was a significant reduction in urine output (mL / kg / h) (Figure 48) compared to basal levels (TO). In addition, the expression of the renal tubule injury biomarkers cystatin C and KIM-1 in both serum (Figures 49A and 50A, respectively) and urine samples (Figures 49B and 50B, respectively) increased compared to basal levels (TO). (Serum and urinary creatinine, cystatin C, and KIM-1 levels were measured by ELISA assay (n=6 independent samples per time point and group), and urine output (ml / kg / h) was recorded for each animal. Results are presented as mean±SEM in Figures 45D, 46D, 46E, and 48. Significant differences were assessed by one-way ANOVA with Tukey's correction (ns: p>0.05, *p<0.05 vs. LPS group). **p<0.005, **p<0.0005; §p<0.05, §§p<0.0005 vs. CER20 group). In Figure 45E, Figure 47, and Figures 49A-50B, gray bands indicate two infusions (0-1 hour and 3-4 hours) of saline or CER-001 at a flow rate of 250 ml / h. Results are presented as mean ± SEM. Significant differences were assessed using two-way repeated measures ANOVA with Tukey's correction (ns: p>0.05, *p<0.05 **p<0.005, **p<0.0005 vs. LPS group; §p<0.05, §§p<0.0005 vs. CER20 group).

[0500] Overall, these results confirmed renal injury in endotoxemic pigs, in contrast to pigs receiving CER-001 treatment, which maintained creatinine levels, urine volume, and serum / urinary cystatin C and KIM-1 expression at median baseline levels (T0) compared to control animals.

[0501] This renal injury was highlighted by significant morphological changes in the renal parenchyma, including desquamation, tubular vacuolization, epithelial flattening, necrosis, inflammatory cell infiltration, significant fibrin deposition, reduced capillary numbers in numerous glomeruli, Bowman's capsule dilation (black arrow), and interstitial inflammatory infiltrates (blue arrow). Compare Figure 46A (LPS group) with Figures 46B and 46C (CER-001 group). These observations were reflected by histopathological scores, which showed significantly less renal damage in the treatment groups, especially after two doses of the ApoA-I complex (tubular pathology, Figure 46D; glomerular pathology, Figure 46E; tubular and glomerular pathology scores were obtained as described in the Methods section (n = 6 for each group).

[0502] As a synthetic HDL containing ApoA-I, the potential positive mechanism of action of CER-001 in sepsis can be attributed, on the one hand, to its ability to reduce inflammatory cytokines (see above) and, on the other hand, to its ability to counteract the effects of LPS through direct interaction, as previously described for HDL. Indeed, consistent with the positive effect of CER-001, serum LPS levels measured by ELISA assay were observed to be reduced in treated animals (n = 6 independent samples per time point and group) (Figure 51). The effect was particularly evident after the second injection of CER-001 (T6, CER20x2 vs. LPS, p = 0.0015), with a sustained effect up to 24 hours after infection in both treatment groups.

[0503] One hypothesis for such LPS reduction could be the result of accelerated catabolism / excretion induced by the scavenging effect of CER-001 on LPS, as described for HDL. Because the natural catabolism of HDL allows cholesterol excretion via bile, with HDL being excreted by the liver as its final target, it is hypothesized that the formed LPS-CER-001 complex could rapidly excrete LPS into bile via the liver. Indeed, the amount of LPS measured in the liver was higher for the LPS group than for both CER-001-treated groups, as shown by representative Western blots (Figure 52A) and densitometric analysis (Figure 52B) of LPS and β-actin protein expression. Figure 53 shows a dose-dependent increase in endotoxin in the bile of CER-001-treated septic pigs, as measured by ELISA assay. Interestingly, the time course of human ApoA-I serum levels (Figure 54) and the dose-dependent increase in human ApoA-I in bile samples (Figure 55) confirmed the role of CER-001 in this increased LPS excretion. Human ApoA-I levels were measured by ELISA. Gray bands indicate two injections (0-1 h and 3-4 h) of saline or CER-001 at a flow rate of 250 ml / h. Data are presented as mean ± SEM. Statistically significant differences were assessed by one-way ANOVA with Tukey's correction (ns: p>0.05).

[0504] 7.4. Example 4: Lipid-binding protein molecular therapy in a model of LPS-induced vascular endothelial injury The ability of CER-001, an ApoA-I-containing complex, to attenuate sepsis-induced injury to the vascular endothelium was evaluated in a lipopolysaccharide (LPS)-induced in vitro model.

[0505] 7.4.1 Materials and Methods 7.4.1.1.Cell culture Human umbilical vein endothelial cells (HUVECs, ECs) were purchased from the American Type Culture Collection (ATCC-LGC Standards Srl, Sesto San Giovanni, Milan, Italy). ECs were maintained in their recommended medium, EndGro (Merck Millipore, Darmstadt, Germany).

[0506] Peripheral blood mononuclear cells (PBMCs) were isolated from buffy coats of healthy donors (selected from a research repository) by gradient centrifugation using the Ficoll-Hypaque method as previously described (Sallustio, et al., 2021, Nephrol Dial Transplant 36, 452-464). PBMCs were maintained in their recommended medium (ibid.).

[0507] When cells became confluent, they were stimulated with 0.3 μg / ml, 4 μg / ml LPS (E. coli O111:B4, Sigma-Aldrich, Milan, Italy) and 50, 100, and 500 μg / ml CER-001 for the indicated periods.

[0508] PBMC culture supernatants were collected and analyzed for TNF-α by ELISA (R&D Systems, Minneapolis, MN, USA).

[0509] 7.4.1.2. Cell proliferation assay ECs and PBMCs were incubated with LPS at 0.3 μg / ml and / or CER-001 at 50 and 500 μg / ml for 60 minutes and 24 hours. The proliferation rate was measured by MTT cell proliferation assay kit according to the manufacturer's instructions (Sigma Aldrich). Briefly, 3 × 10 4 Cells / well were seeded into a 96-well plate, and then the cells were treated with LPS and CER-001 as indicated. The absorbance at 570 nm was then measured by a spectrophotometer.

[0510] 7.4.1.3. Immunophenotypic analysis After stimulation, ECs were permeabilized with an IntraPrep kit (Instrumentation Laboratory) and incubated with unconjugated primary antibody p-ENOS (Abcam) for 25 minutes at 4° C. Cells were then washed and labeled with secondary antibody AlexaFluor 488 (Molecular Probes) for 25 minutes at 4° C. Finally, cells were washed twice, resuspended in FACS buffer, and acquired.

[0511] PBMCs were stained with the following monoclonal antibody, CD14 monoclonal antibody (61D3)-PE (eBioscience™, Thermo Fisher Scientific, Italy) for 20 minutes at room temperature in the dark, washed twice, and resuspended in FACS buffer. The stained PBMCs were then harvested.

[0512] Data were acquired using a FC500 (Beckman Coulter) flow cytometer and analyzed with Kaluza software. Three independent studies were performed on both EC and PBMC. Positive areas were measured using isotype-matched mAbs, and a total of 104 events were acquired for each sample.

[0513] 7.4.1.4.Statistical analysis Data shown are representative of three independent studies. Data are presented as mean ± standard deviation (SD) and compared by Student's t-test.

[0514] 7.4.2.Results The effects of LPS and CER-001 on endothelial cells and endothelial nitric oxide synthase (eNOS) activation were analyzed. The MTT cell viability assay results in Figure 58 showed a slight reduction in proliferation after LPS stimulation. CER-001 at 50 and 500 μg / ml did not affect endothelial viability. Endothelial cells treated with LPS and CER-001 at both 50 and 500 μg / ml increased proliferation rates compared to LPS-stimulated cells, especially at the highest concentration.

[0515] eNOS production has been described as a marker of vascular endothelial integrity (Zhao, et al., 2015, J Pharmacol Sci 129, 83-94). In this in vitro model, eNOS phosphorylation and activation (Figures 59-60) were altered by LPS and upregulated by CER-001. Specifically, a strong decrease in eNOS (phospho-S1177) (p-ENOS) was observed 60 minutes after LPS stimulation compared to basal and VEGF (positive control). CER-001 supplementation at 500 μg / ml completely reversed the LPS effect. (Figure 60 shows representative data from one of three studies. The histogram indicates p-ENOS expression levels.)

[0516] In addition, CER-001 modulated the response of peripheral blood mononuclear cells (PBMCs) stimulated for 24 hours with LPS at 0.3 μg / ml and / or CER-001 at 50 and 500 μg / ml, reducing mCD14 expression and TNF-α secretion. As shown in Figure 61, MTT assays showed no significant differences in cell viability relative to baseline for the above conditions. PBMC culture supernatants were analyzed by ELISA, and the results are shown in Figure 62. 24 hours after LPS stimulation, PBMCs increased TNF-α synthesis. Stimulation of PBMCs with CER-001 alone at 50 and 500 μg / ml did not affect TNF-α production. Addition of CER-001 at both 50 and 500 μg / ml to the culture medium of LPS-activated PBMCs reversed the LPS effect. FACS also demonstrated a strong upregulation of CD14 surface expression by PCMB 24 hours after LPS stimulation (Figure 63). PBMCs treated with a combination of LPS and CER-001 maintained CD14 expression at basal levels (Figure 64).

[0517] 7.5. Example 5: CER-001 in Subjects with Septic Shock 7.5.1. Purpose A clinical Phase 2B / 3 trial is being conducted to evaluate CER-001 plus standard of care (SOC) versus placebo plus SOC for 90-day survival in subjects with septic shock. Secondary objectives include observing the effects of CER-001 on organ dysfunction and organ support use, morbidity and mortality, and health-related quality of life, and further evaluating the pharmacokinetics of CER-001 and a series of biomarkers related to CER-001's mode of action.

[0518] Endpoints The primary endpoint is all-cause mortality at 90 days (defined as the fraction of subjects who died regardless of cause). Secondary endpoints include ICU-free days up to 90 days, mechanical ventilation-free days up to 90 days, renal replacement therapy (RRT)-free days up to 90 days, vasopressor-free days up to 30 days, organ support-free days up to 90 days, and sepsis support index score over 90 days.

[0519] The Sepsis Support Index (SSI) is a composite endpoint reflecting organ dysfunction or death within the first 14 days of follow-up. More specifically, within the first 14 days of follow-up, every day on which vasopressors or mechanical ventilation are used, or on which renal dysfunction (defined as a renal Sequential Organ Failure Assessment (SOFA) score of 4) is evident, or on which the patient dies is counted. The sum of the counted days over the 14-day follow-up period defines the SSI score, which can have a maximum of 14. Further explanation of how to calculate the SSI is provided by Geven, et al., 2019, BMJ Open 0:e024475.

[0520] Secondary efficacy endpoints include: Organ dysfunction (Single-day Organ Failure Assessment (SOFA) score over 30 days) Health-related quality of life (change in utility up to 30 days and up to 90 days based on the EuroQol Group 5-item 5-level (EQ-5D-5L) questionnaire) Mental status (assessed at 30, 60, and 90 days)

[0521] Exploratory endpoints include: Eligible location of stay on days 30, 60, and 90 (ICU, hospital, step-down unit, or home) ● Mean arterial pressure (MAP) until ICU discharge (up to 7 days) Pharmacokinetic response (in a subset of approximately 100 subjects in Part 1 of the study) Arterial oxygen partial pressure to inspired oxygen concentration ratio (PaO2 / FiO2 ratio) Sepsis-related cytokines Endothelial biomarkers

[0522] 7.5.3. Clinical trial design This is a double-blind, randomized, placebo-controlled, two-part adaptive clinical trial.

[0523] The overall trial design includes two parts (Part 1 - Phase 2b and Part 2 - Phase 3) depicted in Figure 69.

[0524] Part 1 has a fixed randomization (1:1:1) to placebo, 10 mg / kg CER-001, or 20 mg / kg CER-001. When 30-day survival data are available from approximately 150 randomized subjects, the results of Part 1 will be used to determine whether the study will transition to Part 2 (enrollment in Part 1 will continue during analysis), eliminate one dosing arm, and allocate participants from this arm to the remaining CER-001 arm.

[0525] The investigational medicinal products (IMPs) used are (1) CER-001 sterile solution for intravenous infusion and (2) placebo: 0.9% sterile sodium chloride solution (250 mL). CER-001 is provided frozen in 20 mL vials, each containing approximately 18 mL of product at a concentration (ApoA-I content) of 8 mg / mL. CER-001 is dosed by body weight. All doses are thawed, then diluted to a volume of 250 mL with normal saline, and administered over a 1-hour period (250 mL / hour) using an infusion pump.

[0526] IMP infusion will be initiated as soon as possible, beginning within 48 hours of ICU admission. To ensure IMP treatment is initiated without delay, informed consent will be obtained as soon as possible in accordance with local regulations.

[0527] IMP is administered twice daily, 12 hours apart, for five consecutive days.

[0528] Study participation for individual subjects is depicted in Figure 70. After day 15, subjects discharged from the hospital to their homes or on step-down treatment will be followed remotely. Survival at day 180 will be assessed by telephone.

[0529] 7.5.4. Inclusion and Exclusion Criteria Subjects were at least 11 years old but under 80 years old, had proven or suspected infection, and had septic shock characterized by hypotension (systolic arterial pressure less than 90 mmHg or mean arterial pressure (MAP) less than 65 mmHg) requiring the use of vasopressors for more than 1 hour despite intravenous fluid resuscitation.

[0530] Exclusion criteria included inability to initiate IMP treatment within 24 hours of initiating vasopressors for septic shock; previous severe sepsis with ICU admission within the past 12 months; hypotension secondary to causes other than sepsis (e.g., major trauma including traumatic brain injury, hemorrhage, burns, or congestive heart failure / cardiogenic shock); chronic mechanical ventilation for any reason in the past 6 months or severe COPD within the past 30 days requiring continuous daily oxygen use; chronic kidney disease that is stage 4 or 5 or requires dialysis for any reason within the past 30 days; receipt of a bone marrow transplant within the past 6 months or receipt of chemotherapy within the past 12 months; known malignancy within the past 12 months, except for basal or squamous cell skin cancer; known pregnancy; presence of a DNR or other decision limiting full treatment taken before obtaining informed consent; previous enrollment in this trial; previous use of an investigational drug product within the past month, or planned or concurrent participation in a clinical trial for any investigational drug or device.

[0531] In addition, after randomization, no more than 24 hours may have elapsed since the initiation of vasopressors for septic shock before the start of the IMP infusion.

[0532] 7.5.5. Statistical methods The primary analysis will compare all subjects treated with the CER-001 dosing regimen from both parts of the trial (pooled together and treated as a single arm) to all subjects in the placebo arm from both parts of the trial. The primary analysis will assess CER-001 superiority using a one-sided 5% significance level test. Analyses will be based on both the modified intention-to-treat (mITT) set and the per-protocol (PP) analysis set, with the mITT considered the primary analysis for determining statistical significance and the PP analysis considered ancillary. The mITT will include all randomized subjects who receive at least one dose of blinded therapy.

[0533] A goal of approximately 850 subjects randomized 1:2 to placebo:CER-001 will provide 80% power to detect a 10% absolute risk reduction or a 20% relative risk reduction, assuming a 40% 90-day mortality rate in subjects with septic shock treated according to standard of care.

[0534] Secondary endpoints aim to support the primary efficacy by further demonstrating treatment effect with an acceptable safety profile. All secondary endpoints will be analyzed using both mITT and PP analysis sets.

[0535] All-cause mortality (days 30 and 180) will be analyzed in the same manner as the primary endpoint. Mortality will be presented graphically by Kaplan-Meier plots. Parameters measured in days and sepsis auxiliary indicators will be analyzed by superiority testing using a two-sided 5% significance level. Endpoints addressing changes in SOFA scores, health-related quality of life, cytokine levels, and endothelial dysfunction markers will be analyzed by analysis of variance (ANOVA) or analysis of covariance (ANCOVA) methods, as appropriate, and presented graphically.

[0536] The safety profile, including adverse events, vital signs, and safety laboratory variables, will be summarized descriptively. Safety analyses will be performed using a safety analysis set. The safety analysis set will include all IMP-treated subjects and will be analyzed according to the actual treatment received.

[0537] ApoA-I concentrations over time are presented in the calculation of pharmacokinetic parameters for days 1 and 5 (first and last days of treatment). Peak and trough levels of apoA-I on days 1-6 with the CER-001 dosing regimen are examined graphically.

[0538] Table 5 provides a schedule of study procedures. [Table 17]

[0539] 7.5.1.Results CER-001 therapy provides therapeutic benefit to subjects with septic shock.

[0540] 7.6. Discussion of Examples Current treatment guidelines for sepsis patients are based on hemodynamic resuscitation, supportive care, and appropriate antibiotic therapy. However, in the most critically ill patients, these measures are insufficient to prevent sepsis-related organ dysfunction and the development of AKI. The findings described in the Examples demonstrate the targeted anti-inflammatory effects of CER-001 at the renal and hepatic levels as well as the systemic circulation in a porcine model of endotoxemia. The results indicate that CER-001 enhances the transport of LPS to the liver and promotes its excretion into bile, indirectly attenuating inflammation. Data demonstrated a dose-dependent reduction in endotoxin levels in the serum and liver tissue of treated endotoxemic animals. LPS and ApoA-I levels were increased in bile samples from CER-001-treated animals. The pharmacokinetics of human ApoA-I measured in pig serum from both treatment groups was consistent with the reduced LPS and endothelial and inflammatory biomarker levels mediated by CER-001 treatment. A pilot clinical study testing the safety and efficacy of CER-001 in treating a heterogeneous cohort of patients with sepsis confirmed the ability of CER-001 treatment to enhance LPS clearance, modulate inflammatory responses secondary to sepsis, and prevent endothelial and organ dysfunction.

[0541] 8. Specific Embodiments The present disclosure is illustrated by the numbered embodiments set forth below. 1. A method of treating a subject having or at risk of having a condition, wherein the condition is optionally an acute condition, and the method comprises administering to the subject a dose, e.g., a high dose, of a lipid binding protein molecule. 2. The method of embodiment 1, wherein the condition is associated with abnormal levels of TREM-1, albumin, interleukin-10 (IL-10), kynurenine pathway biomarkers, TNF-α, MCP-1, IL-6, IL-8, VCAM-1, or ICAM-1. 3. The method of embodiment 1 or 2, wherein the pathology is associated with abnormal levels of ApoA-I, eNOS, or CD14. 4. The method of any one of embodiments 1 to 3, wherein the pathology is associated with abnormal levels of TREM-1. 5. The method of any one of embodiments 1 to 4, wherein the subject has above-normal levels of TREM-1 prior to administration of the lipid binding protein molecule. 6. The method of any one of embodiments 1 to 5, further comprising measuring the subject's level of TREM-1 prior to administering the dose. 7. The method of any one of embodiments 1 to 6, wherein the dose comprises an amount of a lipid binding protein molecule that reduces the level of TREM-1 in the subject. 8. The method of any one of embodiments 1-7, wherein the pathology is associated with abnormal levels of albumin. 9. The method of any one of embodiments 1-8, wherein the subject has a below-normal level of albumin prior to administration of the lipid binding protein molecule. 10. The method of any one of embodiments 1-9, further comprising measuring the subject's albumin level prior to administering the dose. 11. The method of any one of embodiments 1-10, wherein the dose comprises an amount of a lipid binding protein molecule that increases the level of albumin in the subject. 12. The method of any one of embodiments 1-11, wherein the pathology is associated with abnormal levels of a kynurenine pathway biomarker. 13. The method of embodiment 12, wherein the kynurenine pathway biomarker is kynurenine. 14. The method of embodiment 12, wherein the kynurenine pathway biomarker is kynurenic acid. 15. The method of embodiment 12, wherein the kynurenine pathway biomarker is 3-hydroxykynurenine. 16. The method of embodiment 12, wherein the kynurenine pathway biomarker is anthranilic acid. 17. The method of embodiment 12, wherein the kynurenine pathway biomarker is 3-hydroxyanthranilic acid. 18. The method of embodiment 12, wherein the kynurenine pathway biomarker is 2-amino-3-carboxymuconic acid-semialdehyde. 19. The method of embodiment 12, wherein the kynurenine pathway biomarker is picolinic acid. 20. The method of embodiment 12, wherein the kynurenine pathway biomarker is quinolinic acid. 21. The method of embodiment 12, wherein the kynurenine pathway biomarker is quinaldic acid. 22. The method of embodiment 12, wherein the kynurenine pathway biomarker is tryptophan. 23. The method of embodiment 12, wherein the kynurenine pathway biomarker is serotonin. 24. The method of embodiment 12, wherein the kynurenine pathway biomarker is xanthurenic acid. 25. The method of embodiment 12, wherein the kynurenine pathway biomarker is formylkynurenine. 26. The method of embodiment 12, wherein the kynurenine pathway biomarker is the kynurenine / tryptophan ratio. 27. The method of any one of embodiments 1-26, wherein the subject has above-normal levels of kynurenine before administration of the lipid binding protein molecule. 28. The method of any one of embodiments 1-27, further comprising measuring the subject's kynurenine level before administering the dose. 29. The method of any one of embodiments 1-28, wherein the dose comprises an amount of a lipid binding protein molecule that reduces the level of kynurenine in the subject. 30. The method of any one of embodiments 1-29, wherein the subject has above-normal levels of kynurenic acid prior to administration of the lipid binding protein molecule. 31. The method of any one of embodiments 1-30, further comprising measuring the subject's level of kynurenic acid before administering the dose. 32. The method of any one of embodiments 1-31, wherein the dose comprises an amount of a lipid binding protein molecule that reduces the level of kynurenic acid in the subject. 33. The method of any one of embodiments 1-32, wherein the subject has above-normal levels of 3-hydroxykynurenine before administration of the lipid binding protein molecule. 34. The method of any one of embodiments 1-33, further comprising measuring the subject's level of 3-hydroxykynurenine before administering the dose. 35. The method of any one of embodiments 1-34, wherein the dose comprises an amount of a lipid binding protein molecule that reduces the level of 3-hydroxykynurenine in the subject. 36. The method of any one of embodiments 1-35, wherein the subject has above-normal levels of anthranilic acid prior to administration of the lipid binding protein molecule. 37. The method of any one of embodiments 1-36, further comprising measuring the subject's level of anthranilic acid before administering the dose. 38. The method of any one of embodiments 1-37, wherein the dose comprises an amount of a lipid binding protein molecule that reduces the level of anthranilic acid in the subject. 39. The method of any one of embodiments 1-39, wherein the subject has above-normal levels of 3-hydroxyanthranilic acid prior to administration of the lipid binding protein molecule. 40. The method of any one of embodiments 1-39, further comprising measuring the subject's level of 3-hydroxyanthranilic acid before administering the dose. 41. The method of any one of embodiments 1-40, wherein the dose comprises an amount of a lipid binding protein molecule that reduces the level of 3-hydroxyanthranilic acid in the subject. 42. The method of any one of embodiments 1-41, wherein the subject has a higher than normal level of 2-amino-3-carboxymuconic acid-semialdehyde before administration of the lipid binding protein molecule. 43. The method of any one of embodiments 1-42, further comprising measuring the subject's level of 2-amino-3-carboxymuconic acid-semialdehyde before administering the dose. 44. The method of any one of embodiments 1-43, wherein the dose comprises an amount of a lipid-binding protein molecule that reduces the level of 2-amino-3-carboxymuconic acid-semialdehyde in the subject. 45. The method of any one of embodiments 1-44, wherein the subject has above-normal levels of picolinic acid before administration of the lipid binding protein molecule. 46. ​​The method of any one of embodiments 1-45, further comprising measuring the subject's level of picolinic acid before administering the dose. 47. The method of any one of embodiments 1-46, wherein the dose comprises an amount of a lipid binding protein molecule that reduces the level of picolinic acid in the subject. 48. The method of any one of embodiments 1-47, wherein the subject has above-normal levels of quinolinic acid before administration of the lipid binding protein molecule. 49. The method of any one of embodiments 1-48, further comprising measuring the subject's level of quinolinic acid before administering the dose. 50. The method of any one of embodiments 1-49, wherein the dose comprises an amount of a lipid binding protein molecule that reduces the level of quinolinic acid in the subject. 51. The method of any one of embodiments 1-51, wherein the subject has above-normal levels of quinaldic acid prior to administration of the lipid binding protein molecule. 52. The method of any one of embodiments 1-51, further comprising measuring the subject's level of quinaldic acid before administering the dose. 53. The method of any one of embodiments 1-52, wherein the dose comprises an amount of a lipid-binding protein molecule that reduces the level of quinaldic acid in the subject. 54. The method of any one of embodiments 1-53, wherein the subject has a below-normal level of tryptophan before administration of the lipid binding protein molecule. 55. The method of any one of embodiments 1-54, further comprising measuring the subject's tryptophan levels before administering the dose. 56. The method of any one of embodiments 1-55, wherein the dose comprises an amount of a lipid binding protein molecule that increases the level of tryptophan in the subject. 57. The method of any one of embodiments 1-56, wherein the subject has a below-normal level of serotonin before administration of the lipid binding protein molecule. 58. The method of any one of embodiments 1-57, further comprising measuring the subject's serotonin levels before administering the dose. 59. The method of any one of embodiments 1-58, wherein the dose comprises an amount of a lipid binding protein molecule that increases the level of serotonin in the subject. 60. The method of any one of embodiments 1-59, wherein the subject has a higher than normal level of xanthurenic acid before administration of the lipid binding protein molecule. 61. The method of any one of embodiments 1-60, further comprising measuring the subject's level of xanthurenic acid before administering the dose. 62. The method of any one of embodiments 1-61, wherein the dose comprises an amount of a lipid-binding protein molecule that reduces the level of xanthurenic acid in the subject. 63. The method of any one of embodiments 1-62, wherein the subject has above-normal levels of formylkynurenine before administration of the lipid binding protein molecule. 64. The method of any one of embodiments 1-63, further comprising measuring the subject's level of formylkynurenine before administering the dose. 65. The method of any one of embodiments 1-64, wherein the dose comprises an amount of a lipid binding protein molecule that reduces the level of formylkynurenine in the subject. 66. The method of any one of embodiments 1-65, wherein the subject has a supranormal kynurenine / tryptophan ratio before administration of the lipid binding protein molecule. 67. The method of any one of embodiments 1-66, further comprising measuring the subject's kynurenine / tryptophan ratio before administering the dose. 68. The method of any one of embodiments 1-67, wherein the dose comprises an amount of a lipid binding protein molecule that reduces the kynurenine / tryptophan ratio in the subject. 69. The method of any one of embodiments 1-68, wherein the pathology is associated with abnormal levels of TNFα. 70. The method of any one of embodiments 1-69, wherein the subject has above-normal levels of TNFα before administration of the lipid binding protein molecule. 71. The method of any one of embodiments 1-70, further comprising measuring the subject's level of TNFα before administering the dose. 72. The method of any one of embodiments 1-71, wherein the dose comprises an amount of a lipid binding protein molecule that reduces the level of TNFα in the subject. 73. The method of any one of embodiments 1-72, wherein the pathology is associated with abnormal levels of MCP-1. 74. The method of any one of embodiments 1-73, wherein the subject has above-normal levels of MCP-1 before administration of the lipid binding protein molecule. 75. The method of any one of embodiments 1-74, further comprising measuring the subject's level of MCP-1 before administering the dose. 76. The method of any one of embodiments 1-75, wherein the dose comprises an amount of a lipid binding protein molecule that reduces the level of MCP-1 in the subject. 77. The method of any one of embodiments 1-76, wherein the pathology is associated with abnormal levels of IL-6. 78. The method of any one of embodiments 1-77, wherein the subject has above-normal levels of IL-6 before administration of the lipid binding protein molecule. 79. The method of any one of embodiments 1-78, further comprising measuring the subject's level of IL-6 before administering the dose. 80. The method of any one of embodiments 1-79, wherein the dose comprises an amount of a lipid binding protein molecule that reduces the level of IL-6 in the subject. 81. The method of any one of embodiments 1-80, wherein the pathology is associated with abnormal levels of IL-8. 82. The method of any one of embodiments 1-81, wherein the subject has above-normal levels of IL-8 before administration of the lipid binding protein molecule. 83. The method of any one of embodiments 1-82, further comprising measuring the subject's level of IL-8 before administering the dose. 84. The method of any one of embodiments 1-83, wherein the dose comprises an amount of a lipid binding protein molecule that reduces the level of IL-8 in the subject. 85. The method of any one of embodiments 1-84, wherein the pathology is associated with abnormal levels of ApoA-I. 86. The method of any one of embodiments 1-85, wherein the subject has a below-normal level of ApoA-I before administration of the lipid binding protein molecule. 87. The method of any one of embodiments 1-86, further comprising measuring the subject's level of ApoA-I before administering the dose. 88. The method of any one of embodiments 1-87, wherein the dose comprises an amount of a lipid binding protein molecule that increases the level of ApoA-I in the subject. 89. The method of any one of embodiments 1-88, wherein the pathology is associated with abnormal levels of eNOS. 90. The method of any one of embodiments 1-89, wherein the subject has a below-normal level of eNOS before administration of the lipid binding protein molecule. 91. The method of any one of embodiments 1-90, further comprising measuring the level of eNOS in the subject before administering the dose. 92. The method of any one of embodiments 1-91, wherein the dose comprises an amount of a lipid-binding protein molecule that increases the level of eNOS in the subject. 93. The method of any one of embodiments 1-92, wherein the pathology is associated with abnormal levels of CD14. 94. The method of any one of embodiments 1-93, wherein the subject has above-normal levels of CD14 before administration of the lipid binding protein molecule. 95. The method of any one of embodiments 1-94, further comprising measuring the subject's CD14 level before administering the dose. 96. The method of any one of embodiments 1-95, wherein the dose comprises an amount of a lipid binding protein molecule that reduces the level of CD14 in the subject. 97. The method of any one of embodiments 1-96, wherein the pathology is associated with abnormal levels of VCAM-1 or ICAM-1. 98. The method of any one of embodiments 1-97, wherein the subject has above-normal levels of VCAM-1 before administration of the lipid binding protein molecule. 99. The method of any one of embodiments 1-98, further comprising measuring the subject's level of VCAM-1 before administering the dose. 100. The method of any one of embodiments 1-99, wherein the dose comprises an amount of a lipid binding protein molecule that reduces the level of VCAM-1 in the subject. 101. The method of any one of embodiments 1-100, wherein the subject has above-normal levels of ICAM-1 before administration of the lipid binding protein molecule. 102. The method of any one of embodiments 1-101, further comprising measuring the subject's level of ICAM-1 before administering the dose. 103. The method of any one of embodiments 1-102, wherein the dose comprises an amount of a lipid binding protein molecule that reduces the level of ICAM-1 in the subject. 104. The method of any one of embodiments 1-103, wherein the pathology is a bacterial infection. 105. The method of embodiment 104, wherein the bacterial infection is a Staphylococcus aureus infection. 106. The method of embodiment 104, wherein the bacterial infection is an Escherichia coli infection. 107. The method of embodiment 104, wherein the bacterial infection is a Streptococcus pneumoniae infection. 108. The method of embodiment 104, wherein the bacterial infection is a Klebsiella pneumoniae infection. 109. The method of embodiment 104, wherein the bacterial infection is a Pseudomonas aeruginosa infection. 110. The method of embodiment 104, wherein the bacterial infection is an Acinetobacter baumannii infection. 111. The method of embodiment 104, wherein the bacterial infection is a Bacteroides fragilis infection. 112. The method of embodiment 104, wherein the bacterial infection is a Klebsiella pneumoniae infection. 113. The method of embodiment 104, wherein the bacterial infection is a Proteus mirabilis infection. 114. The method of embodiment 104, wherein the bacterial infection is a ...

Claims

1. A method of treating a subject having or at risk of having a condition, wherein the condition is optionally an acute condition, the method comprising administering to the subject a dose of a lipid binding protein molecule.

2. 10. The method of claim 1, wherein the condition is associated with abnormal levels of TREM-1, albumin, interleukin-10 (IL-10), a kynurenine pathway biomarker, TNF-α, MCP-1, IL-6, IL-8, VCAM-1, ICAM-1, ApoA-I, eNOS, or CD14.

3. 3. The method of claim 2, wherein the kynurenine pathway biomarker is kynurenine, kynurenic acid, 3-hydroxykynurenine, anthranilic acid, 3-hydroxyanthranilic acid, 2-amino-3-carboxymuconic acid-semialdehyde, picolinic acid, quinolinic acid, quinaldic acid, tryptophan, serotonin, xanthurenic acid, formylkynurenine, or the kynurenine / tryptophan ratio.

4. 4. The method of any one of claims 1-3, further comprising measuring the subject's levels of TREM-1, albumin, kynurenic acid, 3-hydroxykynurenine, anthranilic acid, 3-hydroxyanthranilic acid, 2-amino-3-carboxymuconic acid-semialdehyde, picolinic acid, quinolinic acid, quinaldic acid, tryptophan, serotonin, xanthurenic acid, formylkynurenine, kynurenine / tryptophan ratio, TNFα, MCP-1, IL-6, IL-8, ApoA-I, eNOS, CD14, VCAM-1, or ICAM-1 prior to administering the dose.

5. The pathological conditions include sepsis, sepsis-induced acute kidney injury (AKI), bacterial infection, gram-positive bacterial infection, gram-negative bacterial infection, viral infection, acute myocardial infarction (AMI), Alzheimer's disease, chronic inflammatory bowel disease (IBD), cardiovascular disease (CVD), stroke, transient ischemic attack, cytokine release syndrome (CRS), transplanted organ rejection, ischemia-reperfusion-induced tissue injury, postoperative inflammation, psoriasis, hypoalbuminemia, attention-deficit / hyperactivity disorder (ADHD), central nervous system disorders (CNS), and the like.

5. The method of any one of claims 1 to 4, wherein the condition is a nervous system (CNS) disease, COVID-19 cognitive decline, depression or major depressive disorder, epilepsy, HIV-associated neurocognitive disorder, Huntington's disease, inflammatory bowel disease (IBD), long-term cognitive decline ("brain fog"), death or neurological deficit following cardiac arrest, multiple sclerosis (MS), Parkinson's disease, schizophrenia, liver damage, kidney damage, vascular endothelial damage, or acute respiratory distress syndrome (ARDS).

6. The method of any one of claims 1 to 5, wherein the subject has sepsis.

7. The method of any one of claims 1 to 6, wherein the subject has septic shock.

8. 8. The method of claim 6 or 7, wherein the subject has hypotension requiring the use of vasoconstrictors despite intravenous fluid resuscitation.

9. The method of any one of claims 6 to 8, wherein the subject has a systolic arterial pressure of less than 90 mmHg.

10. 10. The method of any one of claims 6 to 9, wherein the subject has a mean arterial pressure (MAP) of less than 65 mmHg.

11. 11. The method of any one of claims 6 to 10, wherein the subject has hypotension (e.g., systolic arterial pressure less than 90 mmHg or mean arterial pressure (MAP) less than 65 mmHg) requiring the use of vasoconstrictors despite intravenous fluid resuscitation.

12. 12. The method of any one of claims 6 to 11, wherein the subject has hypotension (e.g., systolic arterial pressure less than 90 mmHg or mean arterial pressure (MAP) less than 65 mmHg) requiring the use of vasopressors for more than one hour despite intravenous fluid resuscitation.

13. 9. The method of any one of claims 6 to 8, wherein the subject has hypotension requiring vasoconstrictor therapy to maintain a mean arterial pressure of 65 mmHg or greater.

14. 14. The method of any one of claims 6 to 13, wherein the subject has a serum lactate level greater than 2 mmol / L despite intravenous fluid resuscitation.

15. 15. The method of any one of claims 6 to 14, wherein administration of the lipid binding protein molecule is initiated within one day of the initiation of vasoconstrictor therapy.

16. 15. The method of any one of claims 6 to 14, wherein administration of the lipid binding protein molecule is initiated within 24 hours of the initiation of vasoconstrictor therapy.

17. The method of any one of claims 6 to 16, further comprising administering to the subject a standard of care therapy for sepsis.

18. 18. The method of any one of claims 1 to 17, wherein the doses are administered over a period of from 1 day to approximately 3 weeks.

19. 19. The method of claim 18, wherein the doses are administered over a period of 5 days.

20. 20. The method of any one of claims 1 to 19, wherein multiple individual doses are administered daily or twice daily.

21. 21. The method of claim 20, wherein the multiple individual doses are administered twice daily.

22. 22. The method of claim 21, comprising administering two or more doses approximately 12 hours apart.

23. The method of any one of claims 1 to 22, wherein the lipid-binding protein molecule is an apolipoprotein.

24. 24. The method of claim 23, wherein the apolipoprotein is ApoA-I.

25. 25. The method of claim 24, wherein the ApoA-I has the amino acid sequence of SEQ ID NO:

3.

26. The method of claim 24 or 25, wherein the ApoA-I is recombinant ApoA-I.

27. 27. The method of claim 26, wherein the ApoA-I is produced by a mammalian host cell.

28. 28. The method of claim 27, wherein the mammalian host cell is a CHO cell.

29. 29. The method of any one of claims 1 to 28, wherein the lipid-binding protein molecule is a component of a lipid-binding protein-based complex.

30. 30. The method of claim 29, wherein the lipid-binding protein-based complex is a reconstituted HDL or an HDL mimetic.

31. 31. The method of claim 29 or claim 30, wherein the lipid-binding protein-based complex comprises sphingomyelin.

32. 32. The method of claim 31 , wherein the lipid-binding protein-based complex comprises a synthetic sphingomyelin.

33. 33. The method of claim 32, wherein the sphingomyelin comprises palmitoyl sphingomyelin.

34. 34. The method of any one of claims 29 to 33, wherein the lipid-binding protein-based complex comprises a negatively charged lipid.

35. 35. The method of claim 34, wherein the negatively charged lipid is 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol) (DPPG) or a salt thereof.

36. 31. The method of claim 30, wherein the lipid-binding protein-based complex comprises ApoA-I and phospholipids in an ApoA-I weight:total phospholipid weight ratio of 1:2.7, and comprises the phospholipids sphingomyelin and 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(l-glycerol) (DPPG) in a sphingomyelin:DPPG weight:weight ratio of 97:

3.

37. 31. The method of claim 30, wherein the lipid-binding protein-based conjugate is CER-001, CSL-111, CSL-112, CER-522 ETC-216, or ETC-642.

38. 38. The method of claim 37, wherein the lipid-binding protein-based conjugate is CER-001.

39. 39. The method of any one of claims 1 to 38, wherein the lipid binding protein molecule is administered systemically, optionally by injection.

40. 40. The method of any one of claims 1 to 39, wherein each individual dose of the lipid binding protein molecule administered is 4 to 40 mg / kg (on a protein weight basis).

41. 41. The method of claim 40, wherein each individual dose of said lipid binding protein molecule is 10-20 mg / kg (on a protein weight basis).

42. 41. The method of claim 40, wherein each individual dose of the lipid binding protein molecule is 10 mg / kg (on a protein weight basis).

43. 41. The method of claim 40, wherein each individual dose of the lipid binding protein molecule is 20 mg / kg (on a protein weight basis).

44. 44. The method of any one of claims 1 to 43, wherein two individual doses are administered to the subject per day for five days.

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