Methods for treating cytokine storm-related diseases with recombinant mutant human angiopoietin-like 4

Abstract

The present disclosure provides a novel approach to the prevention, treatment, and alleviation of diseases in patients that occur after, are induced by, or are exacerbated by cytokine storm. More specifically, the present disclosure describes the administration of angiopoietin-like 4 protein (ANGPTL4) to inhibit, prevent, or neutralize the biological action of certain cytokines and related proteins. Also contemplated herein are kits and compositions designed for use in inhibiting, preventing, or neutralizing the biological action of certain cytokines and related proteins.

Description

[Technical field]

[0001] Government funding This invention was made with government support under grants awarded by the National Institutes of Health, including R01DK101637, R01DK128203 and R01DK129522. The Government has certain rights.

[0002] This application claims priority to U.S. Provisional Patent Application No. 63 / 309,754, entitled “Methods of Treating a Cytokine Storm Related Disease with Recombinant Mutated Human Angiopoietin-Like 4,” filed February 14, 2022, the entire contents of which are incorporated herein by reference. [Background technology]

[0003] Cytokine storms may complicate many types of human diseases. For example, viral infections may initiate cytokine production as part of the innate and adaptive immune responses. The extensive cytokine storms recorded early in the COVID-19 pandemic may contribute to organ damage, and a novel evidence-based model of cytokine-mediated end-organ damage has been developed. See Huang et al., "Clinical features of patients infected with 2019 novel coronavirus in Wuhan, China," (2020) Lancet 395:497-506. Among the three organs studied, literature on cardiac involvement shows elevated cardiac troponin I levels (simulating acute myocardial infarction), myocarditis, myocardial necrosis, pericarditis, arrhythmias, and heart failure. See Topol, "COVID-19 can affect the heart," (2020) Science 370:408-409; Inciardi et al., "Cardiac involvement in a patient with coronavirus disease 2019 (COVID-19)" (2020) JAMA Cardiol. 5:819-824. Evidence of liver injury includes increased aminotransferase levels, hepatocellular injury, inflammation, and steatosis. See Herta et al., "COVID-19 and the liver-Lessons learned," (2021) Liver Int. 41 Suppl 1:1-8. Renal manifestations are very common in hospitalized patients with COVID-19, with nearly 40% developing proteinuria and about one-third developing acute kidney injury (AKI).See Cheng et al., "Kidney disease is associated with in-hospital death of patients with COVID-19," (2020) Kidney Int. 97:829-838; Hirsch et al., "Acute kidney injury in patients hospitalized with COVID-19," (2020) Kidney Int. 98:209-218. Kidney biopsy studies in COVID-19 patients with severe proteinuria and / or impaired renal function have shown that the collapsing variant of focal and segmental glomerulosclerosis (FSGS) and acute kidney injury were the most commonly documented. See Kudose et al., "Kidney Biopsy Findings in Patients with COVID-19," (2020) J Am Soc Nephrol. 31:1959-1968; Nasr et al., "Kidney Biopsy Findings in Patients With COVID-19, Kidney Injury and Proteinuria," (2021) Am J Kidney Dis. 77:465-468 (2021).

[0004] The COVID-19 cytokine storm model may offer a mechanistic comparison with rare manifestations of the cytokine storm in the common cold, which shares some elements with renal disease. Basnet et al., “Rhinoviruses and Their Receptors”, (2019) Chest 155:1018–1025; Wine et al., “Cytokine responses in the common cold and otitis media”, (2012) Curr Allergy Asthma Rep.12:574–581; Nieters et al., “Cross-sectional study on cytokine polymorphisms, cytokine production after T-cell stimulation and clinical parameters in a random sample of a German population", (2001) Hum Genet. 108:241-248; Noah et al., "Nasal cytokine production in viral acute upper respiratory infection of childhood", (1995) J Infect Dis. 171:584-592; van Kempen et al., "An update on the pathophysiology of rhinovirus upper respiratory tract" "Rhinology infections", (1999) 37:97~103; Whiteman et al., “IFN-gamma regulation of ICAM-1 receptors in bronchial epithelial cells: soluble ICAM-1 release inhibits human rhinovirus infection”, (2008) J Inflamm(Lond).5:8; Jartti et al., “Systemic T-helper and T-regulatory cell type cytokine responses in rhinovirus vs.See, "respiratory syncytial virus induced early wheezing: an observational study," (2009) Respir Res. 10:85; Hershey et al., "The association of atopy with a gain-of-function mutation in the alpha subunit of the interleukin-4 receptor," (1997) N Engl J Med. 337:1720-1725; Abdel-Hafez et al., "Idiopathic nephrotic syndrome and atopy: is there a common link?", (2009) Am J Kidney Dis. 54:945-953.

[0005] The common cold, often caused by rhinovirus, triggers nearly 70% of recurrent episodes of the glomerular diseases MCD and FSGS. See Passioti et al., "The common cold: potential for future prevention or cure," (2014) Curr Allergy Asthma Rep. 14:413; Takahashi et al., "Triggers of relapse in steroid-dependent and frequently relapsing nephrotic syndrome," (2007) Pediatr Nephrol. 22:232-236. [Prior art documents] [Non-patent literature]

[0006] [Non-Patent Document 1] Huang et al., “Clinical features of patients infected with 2019 novel coronavirus in Wuhan,China”, (2020) Lancet 395:497~506 [Non-Patent Document 2] Topol, “COVID-19 can affect the heart”, (2020) Science 370: pp. 408–409. [Non-Patent Document 3] Inciardi et al., “Cardiac involvement in a patient with coronavirus disease 2019 (COVID-19)” (2020) JAMA Cardiol.5:819-824 [Non-Patent Document 4] Herta et al., "COVID-19 and the liver-Lessons learned", (2021) Liver Int.41 Suppl 1:1~8 [Non-Patent Document 5] Cheng et al., “Kidney disease is associated with in-hospital death of patients with COVID-19”, (2020) Kidney Int.97:829–838 [Non-Patent Document 6] Hirsch et al., "Acute kidney injury in patients hospitalized with COVID-19," Kidney Int. 98:209-218 (2020) [Non-Patent Document 7] Kudose et al., “Kidney Biopsy Findings in Patients with COVID-19”, (2020) J Am Soc Nephrol.31:1959~1968 [Non-Patent Document 8] Nasr et al., "Kidney Biopsy Findings in Patients With COVID-19, Kidney Injury and Proteinuria", (2021) Am J Kidney Dis.77:465-468 (2021) [Non-Patent Document 9] Basnet et al., "Rhinoviruses and Their Receptors," (2019) Chest 155:1018-1025 [Non-Patent Document 10] Wine et al., “Cytokine responses in the common cold and otitis media”, (2012) Curr Allergy Asthma Rep. 12:574–581 [Non-Patent Document 11] Nieters et al., "Cross-sectional study on cytokine polymorphisms, cytokine production after T-cell stimulation and clinical parameters in a random sample of a German population", (2001) Hum Genet. 108: 241-248. [Non-Patent Document 12] Noah et al., "Nasal cytokine production in viral acute upper respiratory infection of childhood", (1995) J Infect Dis. 171: 584-592. [Non-Patent Document 13] van Kempen et al., "An update on the pathophysiology of rhinovirus upper respiratory tract infections", (1999) Rhinology 37:97-103 [Non-Patent Document 14] Whiteman et al., “IFN-gamma regulation of ICAM-1 receptors in bronchial epithelial cells: soluble ICAM-1 release inhibits human rhinovirus infection”, (2008) J Inflamm(Lond).5:8. [Non-Patent Document 15] Jartti et al., “Systemic T-helper and T-regulatory cell type cytokine responses in rhinovirus vs.respiratory syncytial virus induced early wheezing: an observational study”, (2009) Respir Res.10:85 [Non-Patent Document 16] Hershey et al., "The association of atopy with a gain-of-function mutation in the alpha subunit of the interleukin-4 receptor", (1997) N Engl J Med. 337: 1720-1725. [Non-Patent Document 17] Abdel-Hafez et al., “Idiopathic nephrotic syndrome and atopy: is there a common link?” (2009) Am J Kidney Dis.54:945-953 [Non-Patent Document 18] Passioti et al., “The common cold: potential for future prevention or cure”, (2014) Curr Allergy Asthma Rep. 14: 413 pages. [Non-Patent Document 19] Takahashi et al., “Triggers of relapse in steroid-dependent and frequently relapsing nephrotic syndrome”, (2007) Pediatr Nephrol.22:232-236 Summary of the Invention [Problem to be solved by the invention]

[0007] The pathway of this relapse is unclear, but the involvement of cytokine storm remains unclear. Despite efforts to understand the resulting cytokine storm seen after COVID-19 infection, there is still a need to understand the underlying mechanisms of this phenomenon and the resulting clinical symptoms. Such understanding will facilitate the design of therapeutic approaches to reduce cytokine storm-related organ damage. [Means for solving the problem]

[0008] Summary of the Invention The present disclosure relates to a method for preventing or treating the effects of a disease in a patient, comprising administering to the patient a pharmaceutical composition comprising human angiopoietin-like 4 protein (ANGPTL4), wherein ANGPTL4 neutralizes the biological effects of one or more cytokines from the patient, and the disease occurs after, is induced by, or is exacerbated by a cytokine storm. The human angiopoietin-like 4 protein (ANGPTL4) may be a recombinant form of human angiopoietin-like 4 protein (ANGPTL4). In certain embodiments, the human angiopoietin-like 4 protein (ANGPTL4) is a mutated recombinant form of human angiopoietin-like 4 protein (ANGPTL4). In certain embodiments, human angiopoietin-like 4 protein (ANGPTL4) is protein 8520 (SEQ ID NO:5), protein 8496 (SEQ ID NO:6), protein 8501 (SEQ ID NO:7), protein 8506 (SEQ ID NO:8), protein 8511 (SEQ ID NO:9) or protein 8515 (SEQ ID NO:10). In certain embodiments, ANGPTL4 is a polypeptide having an amino acid sequence having at least 90% sequence identity to SEQ ID NO:5 (protein 8520).

[0009] In certain embodiments, human angiopoietin-like 4 protein (ANGPTL4) is capable of inhibiting the biological action of one or more cytokines and related proteins in a patient, the one or more cytokines and related proteins being selected from the group including IL-2, IL-4, soluble IL-4Rα, IL-6, IL-13, IFN-γ, TNFα, soluble ICAM-1, and ACE-2.

[0010] In certain embodiments of the method, the cytokine storm is a result of a viral infection. In some embodiments, the viral infection is caused by an infection selected from the group consisting of SARS-CoV-1, SARS-CoV-2, rhinovirus, influenza, parainfluenza, respiratory syncytial virus, adenovirus, enterovirus, other coronaviruses, cytomegalovirus (CMV), Epstein-Barr virus (EBV), Middle East respiratory syndrome (MERS), and Ebola virus.

[0011] In certain embodiments of the method, the disease is induced by sepsis, primary or secondary hemophagocytic lymphohistiocytosis (HLH), an autoinflammatory disorder, group A streptococcus, bacteria, fungi, tumors or other cancers, organ transplants, diabetes, or metabolic syndrome.

[0012] In certain embodiments, the disease is a viral disease, hi certain embodiments, the viral disease is SARS-CoV-1, SARS-CoV-2, rhinovirus, influenza, parainfluenza, respiratory syncytial virus, adenovirus, enterovirus, other coronaviruses, cytomegalovirus (CMV), Epstein-Barr virus (EBV), Middle East respiratory syndrome (MERS), or Ebola virus.

[0013] In certain embodiments, the disease affects the heart, liver and kidneys following the onset or worsening of a cytokine storm, hi other embodiments, these phenomena are acute kidney injury, myocarditis, pericarditis, cardiac damage, hepatitis and liver damage, and the worsening of glomerular diseases such as minimal change disease (MCD) and focal segmental glomerulosclerosis (FSGS).

[0014] Also disclosed herein is a method for inhibiting acute recurrence or exacerbation of glomerular disease in a patient, the disease occurring after, being induced by, or being exacerbated by a cytokine storm, the method comprising administering to the patient a pharmaceutical composition comprising human angiopoietin-like 4 protein (ANGPTL4), the ANGPTL4 inhibiting or neutralizing the biological effects of one or more cytokines in the subject. In certain embodiments, the human angiopoietin-like 4 protein (ANGPTL4) is a recombinant form of human angiopoietin-like 4 protein (ANGPTL4). In certain embodiments, the human angiopoietin-like 4 protein (ANGPTL4) is a mutated recombinant form of human angiopoietin-like 4 protein (ANGPTL4). In certain embodiments, the mutated recombinant form of human angiopoietin-like 4 protein (ANGPTL4) is selected from the group consisting of protein 8520 (SEQ ID NO:5), protein 8496 (SEQ ID NO:6), protein 8501 (SEQ ID NO:7), protein 8506 (SEQ ID NO:8), protein 8511 (SEQ ID NO:9) and protein 8515 (SEQ ID NO:10). In certain embodiments, ANGPTL is a polypeptide having at least 90% sequence identity to SEQ ID NO:5. In certain embodiments, human angiopoietin-like 4 protein (ANGPTL4) is capable of inhibiting the effects of circulating levels of one or more cytokines and related proteins in a patient.

[0015] In certain embodiments, the glomerular disease comprises nephrotic syndrome. In certain embodiments, the glomerular disease comprises minimal change disease, focal segmental glomerulosclerosis, membranous nephropathy, membranoproliferative glomerulonephritis, diabetic nephropathy, or lupus nephritis. In certain embodiments, the glomerular disease comprises acute relapse or exacerbation of chronic kidney disease caused by minimal change disease, focal segmental glomerulosclerosis, membranous nephropathy, membranoproliferative glomerulonephritis, diabetic nephropathy, or lupus nephritis.

[0016] In certain embodiments, the patient is in complete or partial remission of the glomerular disease prior to the cytokine storm. In certain embodiments, the cytokine storm is induced by a viral infection. In certain embodiments, the viral infection comprises rhinovirus, human coronavirus, adenovirus, respiratory syncytial virus, non-rhinovirus enterovirus, parainfluenza virus, metapneumovirus, influenza, or a combination thereof. In certain embodiments, the administration occurs between about 1 hour and about 7 days after the onset of the cytokine storm. In certain embodiments, the administration occurs within about 7 days after the onset of the cytokine storm. In certain embodiments, the administration comprises parenteral administration. In certain embodiments, the administration comprises intravenous, intraarterial, intramuscular, intradermal, subcutaneous, or intraperitoneal administration. In certain embodiments, the administration comprises intravenous administration.

[0017] The following detailed description of the disclosure may be better understood, by way of example only, with reference to the following drawings, in which: [Brief description of the drawings]

[0018] [Figure 1A] Figures 1A-1I show the effect of cytokine storm in a COVID-19 model. Figure 1A is a flow chart depicting the cytokine storm model in a COVID-19 model causing multisystem organ damage. [Figure 1B] 1 is a chart listing the cytokine components of five different cocktails intended to mimic either the common cold or coronavirus. [Figure 1C] FIG. 11 is a chart showing that baseline (i.e., day 0) urinary albumin / creatinine ratios were similar between BALB / c mice treated with ANGPTL4 or control (human albumin), whereas 24 hours after treatment, urinary albumin / creatinine ratios were significantly lower (P<0.05) in the ANGPTL4-treated group compared to the control-treated group. [Figure 1D] Electron microscopy images are shown demonstrating significantly improved morphological changes in kidney glomeruli and tubules in ANGPTL4-treated mice compared to human albumin-treated controls. [Figure 1E] 1 is a table listing the histological changes in kidney glomeruli and tubules in ANGPTL4-treated mice compared to human albumin-treated controls. The histological changes were significantly ameliorated in ANGPTL4-treated mice. [Figure 1F] 1 shows histological images of the heart in ANGPTL4-treated BALB / C mice compared to human albumin-treated controls. Histological changes were significantly ameliorated in ANGPTL4-treated mice. [Figure 1G] 1 is a chart listing morphological changes in cardiac tissue from ANGPTL4-treated mice compared to human albumin-treated controls. Morphological changes were significantly improved in ANGPTL4-treated mice. [Figure 1H] 1 shows histological images of the liver in ANGPTL4-treated BALB / c mice compared to human albumin-treated controls. Histological changes were significantly ameliorated in ANGPTL4-treated mice. [Figure 1I] 1 is a table listing morphological changes in the livers of ANGPTL4-treated mice compared to human albumin-treated controls. Morphological changes were significantly ameliorated in ANGPTL4-treated mice. [Figure 2A]Figures 2A-2D show the effects of cytokine storm in primary glomerular diseases such as minimal change disease (MCD) and focal segmental glomerulosclerosis (FSGS). Figure 2A is a flow chart depicting the cytokine storm model in a cold model causing multisystem organ damage. [Figure 2B] This chart shows that baseline (i.e., day 0) urinary albuminuria was similar between BALB / cJ mice treated with ANGPTL4 or control (human albumin), whereas 24 hours after treatment, urinary albuminuria was not significantly increased in the ANGPTL4-treated group but was significantly elevated in the control-treated group (P<0.01). [Figure 2C] Electron microscopy images are shown demonstrating that glomerular morphology was significantly better preserved in ANGPTL4-treated BALB / cJ mice compared to human albumin-treated controls. [Figure 2D] 1 is a chart listing changes in glomerular morphology in ANGPTL4-treated BALB / cJ mice compared to human albumin-treated controls. Morphological changes were significantly ameliorated in ANGPTL4-treated mice. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0019] Applicants herein propose the use of proteins that improve and protect the integrity of blood vessels in organs to prevent leakage of circulating proteins and inflammatory cells that can cause damage to target organs. Applicants have identified angiopoietin-like 4 (ANGPTL4) (wild-type and mutant human proteins) as having potent anti-apoptotic effects that may help maintain vascular integrity and thereby prevent leakage of circulating pro-damage proteins and cells. In areas of existing damage, ANGPTL4 can keep repair vessels open to promote healing. Recombinant human mutant ANGPTL4 has many advantages over the wild-type protein, including a longer half-life, no induction of hypertriglyceridemia, and no side effects that may be associated with ANGPTL4 fragments.

[0020] Applicants herein propose the administration of recombinant mutant human ANGPTL4 (e.g., protein 8520 and other previously known mutants) to treat cytokine storm-associated diseases such as viral infections (SARS-CoV-2 / COVID-19, cold and other respiratory viruses), numerous viruses, bacteria, fungi, and other microorganisms that cause cytokine-releasing human diseases, tumors and cancers, and numerous other human conditions involving cytokine release.

[0021] The present disclosure provides a novel approach to the prevention, treatment and alleviation of diseases in patients that occur after, are induced by, or are exacerbated by cytokine storm. More specifically, the present disclosure describes the administration of angiopoietin-like 4 protein (ANGPTL4) to inhibit, prevent, or neutralize the biological action of certain cytokines and related proteins. Also contemplated herein are kits and compositions designed for use in inhibiting, preventing, or neutralizing the biological action of certain cytokines and related proteins.

[0022] Throughout this disclosure, various quantities, such as total amounts, sizes, dimensions, ratios, etc., are presented in the form of ranges. It is to be understood that the description of quantities in the form of ranges is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of any embodiment. Thus, the description of a range should be considered to have specifically disclosed all individual numerical values ​​as well as all possible subranges within that range, unless the context clearly dictates otherwise. For example, the description of a range such as 1-6 should be considered to have specifically disclosed each individual value within that range, such as 1.1, 2, 2.3, 4.62, 5, and 5.9, as well as subranges such as 1-3, 1-4, 1-5, 2-4, 2-6, 3-6, etc. This applies regardless of the breadth of the range. The upper and lower limits of these intervening ranges may independently be included in the smaller ranges, which are also included within the scope of the disclosure, subject to any specifically excluded limit within the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the disclosure unless the context clearly indicates otherwise.

[0023] The terms used herein are merely for the purpose of describing particular embodiments and are not intended to limit any of the embodiments. The singular forms "a", "an" and "the" as used herein are intended to include the plural forms as well, unless the context clearly dictates otherwise. It is further understood that the terms "includes", "comprises", "including" and / or "comprising" as used herein specify the presence of stated features, integers, steps, operations, elements, and / or components, but do not exclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and / or groups thereof. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items. Additionally, it will be understood that an item included in a list in the form "at least one of A, B, and C" may mean (A);(B);(C);(A and B);(B and C);(A and C); or (A, B, and C). Similarly, an item included in a list in the form "at least one of A, B, or C" may mean (A);(B);(C);(A and B);(B and C);(A and C); or (A, B, and C).

[0024] The term "about," as used herein in reference to a numerical value or numerical range, unless specifically stated otherwise or apparent from the context, is understood to mean the stated numerical value and a numerical value of + / -10% of that numerical value, or a recited value in relation to a range, 10% below the recited lower limit and 10% above the recited upper limit.

[0025] The term "patient," as used herein, refers to a human.

[0026] The term "inhibit," as used herein, means to prevent, delay, and / or reduce the effect or likelihood of something happening.

[0027] The term "pharmaceutical acceptable derivative" as used herein means any pharmaceutical acceptable salt, ester, salt of ester, solvate or other derivative of the ANGPTL4 polypeptide or polypeptide derivative of the present disclosure that can provide (directly or indirectly) the function of wild-type ANGPTL4 when administered to a subject, and in certain embodiments, the ANGPTL4 polypeptide or polypeptide derivative exhibits reduced LPL inhibitory activity that is resistant to cleavage. Particularly preferred derivatives are those that, when the ANGPTL4 polypeptide or polypeptide derivative of the present disclosure is administered to a subject (e.g., by allowing an orally administered compound to be more easily absorbed into the blood), increase the bioavailability of such polypeptide, enhance the delivery of such polypeptide to a given biological compartment, increase the solubility to allow administration by injection, modify metabolism, or modify the frequency of excretion. In one embodiment, the derivative is a prodrug.

[0028] The term "pharmaceutically acceptable salts", as used herein, unless otherwise specified, includes salts of acidic or basic groups that may be present in the ANGPTL4 polypeptides or polypeptide derivatives of the disclosure.

[0029] The terms "prevent" or "preventing" as used herein refer to minimizing, reducing or inhibiting the risk of developing a disease state or a parameter related to a disease state or progression or other abnormal or deleterious condition.

[0030] The term "associated protein" with respect to a cytokine, as used herein, refers to a protein that is induced by, capable of binding to, or otherwise associated with the release of the cytokine. As an example, angiotensin-converting enzyme 2 (ACE-2), which SARS-CoV-2 binds to on the cell surface resulting in cytokine release, is one such "associated protein" contemplated by this disclosure.

[0031] The term "sequence identity", as used herein, unless otherwise specified, refers to a comparison between pairs of nucleic acid or amino acid molecules, i.e., the correlation between two amino acid sequences or two nucleotide sequences. Generally, sequences are aligned to obtain the highest degree of agreement. Methods for determining sequence identity are known and can be determined by commercially available computer programs that can calculate the percent identity between two or more sequences. A typical example of such a computer program is CLUSTAL. By way of illustration, a polynucleotide having a nucleotide sequence with at least, for example, 90% sequence identity to a reference nucleotide sequence means that the nucleotide sequence of the polynucleotide is identical to the reference sequence, but the polynucleotide sequence may contain, on average, up to 10 point mutations per each 100 nucleotides of the reference nucleotide sequence. In other words, to obtain a polynucleotide having a nucleotide sequence that is at least 90% identical to the reference nucleotide sequence, up to 10% of the nucleotides in the reference sequence may be deleted or replaced with another nucleotide, or up to 10% of the number of nucleotides in the reference sequence may be inserted into the reference sequence. These mutations of the reference sequence may be present at the 5' or 3' terminal position of the reference nucleotide sequence or anywhere between these terminal positions, either individually interspersed among the nucleotides in the reference sequence or in one or more contiguous groups within the reference sequence. Similarly, a polypeptide having an amino acid sequence with at least, for example, 90% sequence identity with a reference amino acid sequence is intended to mean that the amino acid sequence of the polypeptide is identical to the reference sequence, but the polypeptide sequence may contain, on average, up to 10 amino acid changes per each 100 amino acids of the reference amino acid. In other words, up to 10% of the amino acid residues in the reference sequence may be deleted or replaced with another amino acid, or up to 10% of the number of amino acids of the total amino acid residues in the reference sequence may be inserted into the reference sequence to obtain a polypeptide having an amino acid sequence at least 90% identical to the reference amino acid sequence.These alterations of the reference sequence may occur at the amino or carboxy terminal positions of the reference amino acid sequence or anywhere between these terminal positions, either individually interspersed among residues in the reference sequence or in one or more contiguous groups within the reference sequence.

[0032] The term "therapeutically effective amount," as used herein, refers to an amount of ANGPTL4, either alone or as part of a pharmaceutical composition, capable of producing any detectable positive effect on any symptom, aspect, or characteristic of a disease or condition. Such an effect need not be absolute to be beneficial.

[0033] The term "treat" or "treating" as used herein includes arresting, slowing, halting, or reversing the progression or severity of an existing condition or disorder.

[0034] The term "ANGPTL4" refers herein to human angiopoietin-like 4. Exemplary forms of ANGPTL4 include, but are not limited to, mutant forms of human angiopoietin-like 4, such as protein 8520 (SEQ ID NO:5), protein 8496 (SEQ ID NO:6), protein 8510 (SEQ ID NO:7), protein 8506 (SEQ ID NO:8), protein 8511 (SEQ ID NO:9), or protein 8515 (SEQ ID NO:10). ANGPTL4 is a polypeptide encoded by the ANGPTL4 gene in humans. This gene is a member of the angiopoietin / angiopoietin-like gene family and encodes a glycosylated secreted protein with an N-terminal signal sequence, a coiled-coil domain, a linker region, and a fibrinogen C-terminal domain. This gene is induced under hypoxic conditions in endothelial cells and is a target of peroxisome proliferation activators. The encoded protein is a serum hormone directly involved in regulating glucose homeostasis, lipid metabolism, and insulin sensitivity, and also acts as an apoptotic survival factor for vascular endothelial cells. Alternatively spliced ​​transcript variants encoding different isoforms have been described. The gene was previously named ANGPTL2 but has been renamed ANGPTL4.

[0035] Inhibition of cytokine action In any of the disclosed embodiments, the biological effects of one or more cytokines and associated proteins that are inhibited or neutralized include IL-2, IL-4, soluble IL-4Rα, IL-6, IL-13, IFN-γ, TNFα, soluble ICAM-1, and ACE-2. It is understood that in accordance with the disclosure described herein, depending on the severity of the viral infection or other condition being treated, inhibiting or neutralizing the biological effects of more than one cytokine and / or associated protein may be more effective than inhibiting or neutralizing the effect of a single cytokine and / or associated protein.

[0036] The present invention contemplates that the biological effects of various cytokines and / or associated proteins may be neutralized or inhibited by administration of ANGPTL4. For example, as described herein, the biological effects of cytokines and / or associated proteins may be neutralized or inhibited by administration of a therapeutically effective amount of an agent, the agent including ANGPTL4.

[0037] ANGPTL4 may be administered once a day, more than once a day, or once a week. ANGPTL4 may be administered to a subject by any conventional means, such as orally, intramuscularly, intraperitoneally, or intravenously. When injected, ANGPTL4 may be injected at a single site per dose or at multiple sites per dose. SEQ ID NOs: 1 and 2 show the amino acid and cDNA sequences from human (protein variant 1 isoform a, long form), whereas SEQ ID NOs: 3 and 4 show the amino acid and cDNA sequences from human (protein variant 3 isoform b, short form). SEQ ID NO: 5 shows the amino acid and nucleic acid substitutions of mutant human ANGPTL4 (protein 8520). SEQ ID NO: 6 shows the amino acid and nucleic acid substitutions of mutant human ANGPTL4 (protein 8496). SEQ ID NO: 7 shows the amino acid and nucleic acid substitutions of mutant human ANGPTL4 (protein 8501). SEQ ID NO: 8 shows the amino acid and nucleic acid substitutions of mutant human ANGPTL4 (protein 8506). SEQ ID NO: 9 shows the amino acid and nucleic acid substitutions of mutant human ANGPTL4 (protein 8511). SEQ ID NO: 10 shows the amino acid and nucleic acid substitutions of mutant human ANGPTL4 (protein 8515).

[0038] composition The useful compositions of the present disclosure may comprise the ANGPTL4 described herein, which is useful in the treatment and prevention methods of the present disclosure. The disclosed compositions may comprise one or more of such compounds in combination with a pharmaceutically acceptable carrier. Examples of such carriers and formulation methods can be found in Remington: The Science and Practice of Pharmacy (20th edition, Lippincott, Williams & Wilkins, editor Daniel Limmer) and are known to those skilled in the art. To form a pharmaceutically acceptable composition suitable for administration, such compositions contain a therapeutically effective amount of the compound.

[0039] The pharmaceutical compositions of the present disclosure may be used in the treatment and prevention methods of the present disclosure. Such compositions are administered to a subject in an amount sufficient to deliver a therapeutically effective amount of the compound to be effective in the treatment and prevention methods disclosed herein. The therapeutically effective amount may vary according to various factors, including but not limited to the condition, weight, sex, and age of the subject. Other factors include the mode and site of administration. The pharmaceutical composition may be provided to the subject by any method known in the art. Exemplary routes of administration include but are not limited to subcutaneous, intravenous, topical, epicutaneous, oral, intraosseous, and intramuscular. The compositions of the present disclosure may be administered to a subject only once, or may be administered to a subject more than once. Furthermore, when the composition is administered to a subject more than once, various regimens may be used, such as but not limited to once a day, once a week, or once a month. The composition may also be administered to a subject more than once a day. The therapeutically effective amount and appropriate dosage regimen may be identified by routine testing to obtain optimal activity while minimizing any possible side effects. In addition, co-administration or sequential administration of other agents may be desirable.

[0040] The compositions of the present disclosure may be administered systemically, for example by intravenous administration, or locally, for example by subcutaneous injection or by application of a paste or cream.

[0041] In one embodiment, a nucleic acid is provided that encodes the ANGPTL4 polypeptide or ANGPTL4 polypeptide variant of the present disclosure, which may be in the form of a suitable plasmid or vector. Such a nucleic acid is introduced into a cell, and such a cell can be obtained from a subject by a suitable method known in the art (e.g., electroporation). In one embodiment, the cell is an adipocyte. The cell can be assayed for expression of an ANGPTL4 polypeptide or polypeptide derivative (in one embodiment, expression of a polypeptide can be determined by the presence of a tag on the polypeptide, as discussed herein). The cell expressing the ANGPTL4 polypeptide or polypeptide derivative can then be introduced into the subject. In one embodiment, the cell is administered to the subject by subcutaneous injection, and other administration methods such as those discussed herein can also be used. The cell then expresses the ANGPTL4 polypeptide or ANGPTL4 polypeptide derivative, which is taken up into the circulation.

[0042] The compositions of the present disclosure may further comprise agents that improve the solubility, half-life, absorption, etc. of the compound. Additionally, the compositions of the present disclosure may further comprise agents that attenuate undesirable side effects and / or reduce the toxicity of the compound. Examples of such agents are described in various textbooks, including, but not limited to, Remington: The Science and Practice of Pharmacy (20th Edition, Lippincott, Williams & Wilkins, Editor Daniel Limmer).

[0043] The composition of the present disclosure can be administered in various dosage forms for administration.For example, the composition can be administered in the form of, for example, but not limited to, tablets, capsules, sachets, lozenges, troches, pills, powders, granules, elixirs, tinctures, liquids, suspensions, elixirs, syrups, ointments, creams, pastes, emulsions, or solutions for intravenous administration or injection.Other dosage forms include transdermal administration via patch mechanisms or ointments.All of the above can be modified to provide delayed release and / or sustained release formulations.

[0044] In the present disclosure, the pharmaceutical composition may further comprise a pharma- ceutically acceptable carrier, examples of which include, but are not limited to, vehicles, adjuvants, surfactants, suspending agents, emulsifiers, inert fillers, diluents, excipients, wetting agents, binders, lubricants, buffers, disintegrants and carriers, as well as accessory agents, such as, but not limited to, colorants and flavoring agents (collectively referred to herein as carriers). Typically, a pharma- ceutical acceptable carrier is chemically inert to the active compound and has no adverse side effects or toxicity under the conditions of use. Pharmaceutically acceptable carriers may include polymers and polymer matrices. The nature of the pharma- ceutical acceptable carrier may vary depending on the particular dosage form employed and other characteristics of the composition.

[0045] For oral administration in solid form, such as, but not limited to, tablets, capsules, sachets, lozenges, troches, pills, powders or granules, the compounds may be combined with oral non-toxic pharma- ceutically acceptable inert carriers, such as, but not limited to, inert fillers, suitable binders, lubricants, disintegrants and adjuvants. Suitable binders include, but are not limited to, starch, gelatin, natural sugars, such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums, such as gum arabic, tragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes, and the like. Lubricants used in these dosage forms include, but are not limited to, sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and the like. Disintegrants include, but are not limited to, starch, methylcellulose, agar, bentonite, xanthum gum, and the like. Tablet forms may include one or more of the following: lactose, sucrose, mannitol, corn starch, potato starch, alginic acid, microcrystalline cellulose, gum arabic, gelatin, guar gum, colloidal silicon dioxide, croscarmellose sodium, talc, magnesium stearate, calcium stearate, zinc stearate, stearic acid, as well as other carriers described herein.Lozenge forms may contain the active ingredient in a flavoring agent, usually sucrose and gum arabic or tragacanth, and pastilles contain the active ingredient in an inert base, such as gelatin and glycerin, or sucrose and gum acadia, and emulsions and gels contain, in addition to the active ingredient, carriers known in the art.

[0046] For oral liquid dosage forms, such as, but not limited to, tinctures, solutions, suspensions, elixirs, syrups, the nucleic acid molecules of the present disclosure can be dissolved in diluents, such as water, saline, or alcohol.In addition, oral liquid dosage forms can suitably contain flavored suspending or dispersing agents, such as synthetic and natural gums, such as tragacanth, gum arabic, methylcellulose, and the like.In addition, if desired or necessary, suitable coloring agents or other auxiliary agents can also be incorporated into the mixture.Other dispersing agents that may be employed include glycerin, and the like.

[0047] Formulations suitable for parenteral administration include aqueous and non-aqueous isotonic sterile injection solutions, which may contain aqueous and non-aqueous sterile suspensions that may contain antioxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the patient's blood, suspending agents, solubilizing agents, thickening agents, stabilizers, and preservatives. The compounds may be administered in a physiologically acceptable diluent, for example, in a sterile liquid or mixture of liquids such as water, saline, aqueous dextrose and related sugar solutions, alcohols such as ethanol, isopropanol, or hexadecyl alcohol, glycols such as propylene glycol or polyethylene glycol, e.g., poly(ethylene glycol) 400, glycerol ketals such as 2,2-dimethyl-1,3-dioxolane-4-methanol, ethers, oils, fatty acids, fatty acid esters or glycerides, or acetylated fatty acid glycerides, with or without the addition of a pharma- ceutically acceptable surfactant, for example, but not limited to, a soap, oil or detergent, a suspending agent, for example, but not limited to, pectin, carbomer, methylcellulose, hydroxypropylmethylcellulose, or carboxymethylcellulose, or emulsifying agents and other pharmaceutical adjuvants.

[0048] Oil can be used in parenteral formulation, examples of which include petroleum, animal oil, vegetable oil, or synthetic oil.Specific examples of oil include peanut oil, soybean oil, sesame oil, cottonseed oil, corn oil, olive oil, petrolatum, and mineral oil.Suitable fatty acids for use in parenteral formulation include polyethylene sorbitan fatty acid esters such as sorbitan monooleate, and the high molecular weight adducts of ethylene oxide with hydrophobic bases formed by condensation of propylene oxide with propylene glycol, oleic acid, stearic acid, and isostearic acid.Examples of suitable fatty acid esters are ethyl oleate and isopropyl myristate. Suitable soaps for use in parenteral formulations include fatty alkali metal, ammonium, and triethanolamine salts, and suitable detergents include: (a) cationic detergents, such as dimethyldialkylammonium halides and alkylpyridinium halides; (b) anionic detergents, such as alkyl, aryl, and olefin sulfonates, alkyl, olefin, ether, and monoglyceride sulfates, and sulfosuccinates; (c) nonionic detergents, such as fatty amine oxides, fatty acid alkanolamides, and polyoxyethylene polypropylene copolymers; (d) amphoteric detergents, such as alkyl beta-aminopropionates and 2-alkylimidazoline quaternary ammonium salts; and (e) mixtures thereof.

[0049] Suitable preservatives and buffers may be used in such formulations. To minimize or eliminate irritation at the injection site, such compositions may contain one or more non-ionic surfactants having a hydrophilic-lipophilic balance (HLB) of about 12 to about 17. The amount of surfactant in such formulations ranges from about 5% to about 15% by weight.

[0050] Topical dosage forms containing the nucleic acid molecules of the present disclosure, including but not limited to ointments, creams, pastes, emulsions, etc., can be mixed with various carrier materials well known in the art, such as alcohol, aloe vera gel, allantoin, glycerin, vitamin A and E oil, mineral oil, PPG2 myristyl propionate, etc., to form alcoholic solutions, topical cleansers, cleansing creams, skin gels, skin lotions, and shampoos in the form of cream or gel formulations. Skin exfoliants or skin abrasive preparations can be formulated. Such topical preparations can be applied to patches, dressings or bandages for transdermal delivery, or to bandages or bandages for direct delivery to the site of wounds or skin damage.

[0051] The compounds of the present disclosure can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles.Liposomes can be formed from various phospholipids, such as cholesterol, stearylamine or phosphatidylcholine.Such liposomes can also contain monoclonal antibodies that direct the delivery of liposomes to a specific cell type or group of cell types.

[0052] The compounds of the present disclosure may also be coupled with soluble polymers as targetable drug carriers. Such polymers may include, but are not limited to, polyvinylpyrrolidone, pyran copolymers, polyhydroxypropylmethacryl-amidephenol, polyhydroxyethylaspartamidephenol, or polyethylene oxide polylysine substituted with palmitoyl residues. In addition, the compounds of the present invention may be coupled to crosslinked or amphiphilic block copolymers of biodegradable polymer classes useful in achieving controlled release of drugs, such as polylactide, polyepsiloncaprolactone, polyhydroxybutyrate, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates, and hydrogels.

[0053] Some embodiments of administering an ANGPTL4 polypeptide or derivative to a patient include a form of administration that delivers the polypeptide to the patient's blood. In one example, the polypeptide is administered intravenously. Given the appropriate dosage form, such administration can be performed orally, subcutaneously, or by other means known in the art. The ANGPTL4 polypeptide or derivative can be administered in a therapeutically effective amount, which is generally within a certain range of ratio of the mass of the compound to the mass of the subject. In some embodiments of the method, the polypeptide is administered at a dosage of about 0.005-150,000 μg / kg, 0.5-15,000 μg / kg, 5-1500 μg / kg, or 50-150 μg / kg. Thus, for a typical 70 kg human adult, the dosage can be 0.0035-11,000 mg, 0.035-1100 mg, 0.35-110 mg, or 3.5-11 mg. Administration may be on a regular schedule. In some embodiments of the method, the polypeptide is administered about once every 14 days. In other embodiments, the polypeptide is administered about twice per month. In still other embodiments, the polypeptide is administered about once per month to about twice per month. In further embodiments, the polypeptide is administered once per given period selected from the group consisting of 1 day, 2 days, 3 days, 1 week, 10 days, 2 weeks, 3 weeks, 4 weeks, and 1 month. EXAMPLES

[0054] The following examples are provided for the purpose of illustrating various embodiments of the present invention, and are not meant to limit the present disclosure in any way. The examples of the present invention, together with the methods described herein, are representative of currently preferred embodiments, and are provided merely as examples, and are not intended as limitations on the scope of the present invention. Those skilled in the art will recognize variations therein and other uses that are encompassed within the essence of the present disclosure as defined in the claims. Specifically, the following study illustrates the use of a recombinant mutated form of human angiopoietin-like 4 protein (ANGPTL4) to improve disease outcomes in two different models involving cytokine storm.

[0055] Example 1 Ongoing studies in COVID-19 patients show that viral particles are undetectable in many organs affected by the disease. See references 1-5. This highlights that in hospitalized COVID-19 patients, these organs may be affected by the massive cytokine storm recorded early in the pandemic (Figure 1A). A mutated form of human angiopoietin-like 4 (protein 8520) has previously been shown to reduce proteinuria in rat models of FSGS and diabetic nephropathy. See references 6, 7. Using various elements or innate and adaptive immunity generated during infection, applicants developed a cytokine cocktail that mimics the COVID-19 cytokine storm (Figure 1B) and tested the ability of protein 8520 to reduce glomerular damage in this model. After baseline urine and blood collection, two groups of BALB / c mice (average n=5 mice per group) were intravenously injected with COVID cocktail D at a dose of 1.8x. One hour later, the mice received 10 μg of His-tagged purified protein 8520 or control human albumin. 24 hours after model induction, blood and urine samples were collected and the mice were euthanized, followed by harvesting multiple organs for histological analysis. Urinary albumin was measured by ELISA (Bethyl Labs), and urine and serum creatinine were measured by mass spectrometry. Figure 1C shows that baseline urinary albumin / creatinine ratios were similar between the two groups. At 24 hours, urinary albumin / creatinine ratios were significantly lower in the 8520 treatment compared to the control treatment group (P<0.05). Serum creatinine on day 1 was similar between both groups and was not significantly elevated compared to baseline. Figures 1D and 1E show morphological changes in the glomeruli and tubules of the kidneys in the electron microscope, which were significantly improved in the study (ANGPTL4 8520) group. Figures 1F and 1G show histological changes in the heart that were significantly improved in the ANGPTL4-treated group compared to the control, and Figures 1H and 1I show histological changes in the liver that were significantly improved in the ANGPTL4-treated group compared to the control.

[0056] Example 2 In nearly 70% of patients with primary glomerular diseases such as minimal change disease (MCD) and focal segmental glomerulosclerosis (FSGS), relapses occur after a cold. See Reference 8. In addition, most patients with MCD have a chronic atopic condition. See Reference 9. Previous studies (see Reference 10) have shown the importance of low podocyte ZHX2 expression in the pathogenesis of MCD and FSGS.

[0057] At least half of all colds are caused by rhinovirus infection. Applicant developed a cytokine cocktail that mimics cold-induced relapses of MCD and FSGS (Figures 1B and 2A). When injected intravenously into BALB / cJ mice, the rhinovirus cold cocktail induced a dose-dependent increase in albuminuria, mimicking cold-induced relapses of glomerular disease. Applicant tested the ability of mutant human angiopoietin-like 4 (protein 8520) to reduce albuminuria in this model of mild cytokine storm (see references 6, 7). After collecting baseline blood samples and 18-hour urine in metabolic cages, two groups of BALB / cJ mice were injected intravenously with dose X / 2 of the rhinovirus cold cytokine cocktail. One hour later, the mice received 10 μg of His-tagged purified protein 8520 or control human albumin. Twenty-four hours after model induction, 18-hour urine samples were collected, blood samples were taken, and mice were euthanized, followed by kidney harvest. Urinary albumin was measured by ELISA (Bethyl Labs), and serum creatinine was measured by mass spectrometry. Kidney sections were processed for histology. Figure 2B shows that baseline 18-hour urinary albumin excretion was similar between both groups. At 24 hours, urinary albumin excretion was significantly increased (P<0.01) in the control human albumin-treated group, but not in the 8520-treated group. All mice were normally active 6 and 24 hours prior to euthanasia. Figures 2C and 2D show that glomerular morphology, assessed by electron microscopy, was significantly better preserved in ANGPTL4-treated BALB / cJ mice compared to the human albumin-treated control group.

[0058] References

[0059] [Table 1] TIFF2025506570000002.tif214169

[0060] Although the present invention and its advantages have been described in detail, it is to be understood that various changes, substitutions, and alterations can be made herein without departing from the spirit and scope of the invention, as defined in the appended claims. The methods, assays, and various embodiments thereof described herein are exemplary. Various other embodiments of the methods and assays described herein are possible.

Claims

1. A method for preventing a disease in a patient, comprising administering to the patient a pharmaceutical composition comprising human angiopoietin-like 4 protein (ANGPTL4), wherein ANGPTL4 inhibits or neutralizes the biological action of one or more cytokines from the patient, and the disease occurs after, is induced by, or is exacerbated by a cytokine storm.

2. 2. The method of claim 1, wherein the human angiopoietin-like 4 protein (ANGPTL4) is a recombinant form of human angiopoietin-like 4 protein (ANGPTL4).

3. 2. The method of claim 1, wherein the human angiopoietin-like 4 protein (ANGPTL4) is a mutated recombinant form of human angiopoietin-like 4 protein (ANGPTL4).

4. 4. The method of claim 3, wherein the mutated recombinant form of human angiopoietin-like 4 protein (ANGPTL4) is selected from the group consisting of protein 8520 (SEQ ID NO:5), protein 8496 (SEQ ID NO:6), protein 8501 (SEQ ID NO:7), protein 8506 (SEQ ID NO:8), protein 8511 (SEQ ID NO:9) and protein 8515 (SEQ ID NO:10).

5. The method of claim 3 , wherein ANGPTL is a polypeptide having at least 90% sequence identity to SEQ ID NO:

5.

6. 2. The method of claim 1, wherein human angiopoietin-like 4 protein (ANGPTL4) is capable of inhibiting the effect of circulating levels of one or more cytokines and related proteins in a patient.

7. 7. The method of claim 6, wherein the one or more cytokines and related proteins are selected from the group comprising IL-2, IL-4, soluble IL-4Rα, IL-6, IL-13, IFN-γ, TNFα, soluble ICAM-1 and ACE-2.

8. The method of claim 1 , wherein the cytokine storm is a result of a viral infection.

9. 9. The method of claim 8, wherein the viral infection is caused by an infection selected from the group consisting of SARS-CoV-1, SARS-CoV-2, rhinovirus, influenza, parainfluenza, respiratory syncytial virus, adenovirus, enterovirus, other coronaviruses, cytomegalovirus (CMV), Epstein-Barr virus (EBV), Middle East Respiratory Syndrome (MERS), and Ebola virus.

10. 2. The method of claim 1, wherein the disease is induced by sepsis, primary or secondary hemophagocytic lymphohistiocytosis (HLH), an autoinflammatory disorder, group A streptococcus, bacteria, fungi, tumors or other cancers, organ transplantation, diabetes, or metabolic syndrome.

11. 2. The method of claim 1, wherein the disease is a viral disease.

12. 12. The method of claim 11, wherein the viral disease is selected from the group consisting of SARS-CoV-1, SARS-CoV-2, rhinovirus, influenza, parainfluenza, respiratory syncytial virus, adenovirus, enterovirus, other coronaviruses, cytomegalovirus (CMV), Epstein-Barr virus (EBV), Middle East Respiratory Syndrome (MERS), and Ebola virus.

13. 2. The method of claim 1, wherein the disease is selected from the group including acute kidney injury, myocarditis, pericarditis, cardiac injury, and hepatitis and liver injury.

14. 1. A method for treating a disease in a patient, comprising administering to the patient a pharmaceutical composition comprising human angiopoietin-like 4 protein (ANGPTL4), wherein ANGPTL4 inhibits or neutralizes the action of one or more cytokines from the patient, and the disease occurs after, is induced by, or is exacerbated by a cytokine storm.

15. 15. The method of claim 14, wherein the human angiopoietin-like 4 protein (ANGPTL4) is a recombinant form of human angiopoietin-like 4 protein (ANGPTL4).

16. 15. The method of claim 14, wherein the human angiopoietin-like 4 protein (ANGPTL4) is a mutated recombinant form of human angiopoietin-like 4 protein (ANGPTL4).

17. 17. The method of claim 16, wherein the mutated recombinant form of human angiopoietin-like 4 protein (ANGPTL4) is selected from the group consisting of protein 8520 (SEQ ID NO:5), protein 8496 (SEQ ID NO:6), protein 8501 (SEQ ID NO:7), protein 8506 (SEQ ID NO:8), protein 8511 (SEQ ID NO:9) and protein 8515 (SEQ ID NO:10).

18. 17. The method of claim 16, wherein ANGPTL is a polypeptide having at least 90% sequence identity to SEQ ID NO:

5.

19. 15. The method of claim 14, wherein human angiopoietin-like 4 protein (ANGPTL4) is capable of inhibiting the effect of circulating levels of one or more cytokines and related proteins in a patient.

20. 20. The method of claim 19, wherein the one or more cytokines and related proteins are selected from the group comprising IL-2, IL-4, soluble IL-4Rα, IL-6, IL-13, IFN-γ, TNFα, soluble ICAM-1 and ACE-2.

21. The method of claim 14, wherein the cytokine storm is a result of a viral infection.

22. 22. The method of claim 21, wherein the viral infection is caused by an infection selected from the group consisting of SARS-CoV-1, SARS-CoV-2, rhinovirus, influenza, parainfluenza, respiratory syncytial virus, adenovirus, enterovirus, other coronaviruses, cytomegalovirus (CMV), Epstein-Barr virus (EBV), Middle East Respiratory Syndrome (MERS), and Ebola virus.

23. 15. The method of claim 14, wherein the disease is induced by sepsis, primary or secondary hemophagocytic lymphohistiocytosis (HLH), an autoinflammatory disorder, group A streptococcus, bacteria, fungi, tumors or other cancers, organ transplantation, diabetes, or metabolic syndrome.

24. 15. The method of claim 14, wherein the disease is a viral disease.

25. 25. The method of claim 24, wherein the viral disease is selected from the group consisting of SARS-CoV-1, SARS-CoV-2, influenza, parainfluenza, respiratory syncytial virus, adenovirus, enterovirus, other coronaviruses, cytomegalovirus (CMV), Epstein-Barr virus (EBV), Middle East Respiratory Syndrome (MERS), and Ebola virus.

26. 16. The method of claim 15, wherein the disease is selected from the group including acute kidney injury, myocarditis, pericarditis, cardiac injury, and hepatitis and liver injury.

27. A method for inhibiting acute recurrence or worsening of a glomerular disease in a patient, the disease occurring after, being induced by, or being exacerbated by a cytokine storm, the method comprising administering to the patient a pharmaceutical composition comprising human angiopoietin-like 4 protein (ANGPTL4), wherein ANGPTL4 inhibits or neutralizes the biological action of one or more cytokines in the subject.

28. 28. The method of claim 27, wherein the human angiopoietin-like 4 protein (ANGPTL4) is a recombinant form of human angiopoietin-like 4 protein (ANGPTL4).

29. 28. The method of claim 27, wherein the human angiopoietin-like 4 protein (ANGPTL4) is a mutated recombinant form of human angiopoietin-like 4 protein (ANGPTL4).

30. 30. The method of claim 29, wherein the mutated recombinant form of human angiopoietin-like 4 protein (ANGPTL4) is selected from the group consisting of protein 8520 (SEQ ID NO:5), protein 8496 (SEQ ID NO:6), protein 8501 (SEQ ID NO:7), protein 8506 (SEQ ID NO:8), protein 8511 (SEQ ID NO:9) and protein 8515 (SEQ ID NO:10).

31. 30. The method of claim 29, wherein ANGPTL is a polypeptide having at least 90% sequence identity to SEQ ID NO:

5.

32. 28. The method of claim 27, wherein human angiopoietin-like 4 protein (ANGPTL4) is capable of inhibiting the effect of circulating levels of one or more cytokines and related proteins in a patient.

33. 28. The method of claim 27, wherein the glomerular disease comprises nephrotic syndrome.

34. 28. The method of claim 27, wherein the glomerular disease comprises minimal change disease, focal segmental glomerulosclerosis, membranous nephropathy, membranoproliferative glomerulonephritis, diabetic nephropathy, or lupus nephritis.

35. 28. The method of claim 27, wherein the glomerular disease comprises acute relapse or exacerbation of chronic kidney disease caused by minimal change disease, focal segmental glomerulosclerosis, membranous nephropathy, membranoproliferative glomerulonephritis, diabetic nephropathy, or lupus nephritis.

36. 28. The method of claim 27, wherein the patient is in complete or partial remission of the glomerular disease prior to the cytokine storm.

37. 28. The method of claim 27, wherein the cytokine storm is induced by a viral infection.

38. 38. The method of claim 37, wherein the viral infection comprises rhinovirus, human coronavirus, adenovirus, respiratory syncytial virus, non-rhinovirus enterovirus, parainfluenza virus, metapneumovirus, influenza, or a combination thereof.

39. 28. The method of claim 27, wherein administration occurs between about 1 hour and about 7 days after the onset of the cytokine storm.

40. 28. The method of claim 27, wherein the administration occurs within about 7 days following the onset of the cytokine storm.

41. 28. The method of claim 27, wherein administering comprises parenteral administration.

42. 28. The method of claim 27, wherein administration comprises intravenous, intraarterial, intramuscular, intradermal, subcutaneous or intraperitoneal administration.

43. 28. The method of claim 27, wherein administering comprises intravenous administration.

Images