Compounds and uses thereof

Abstract

Disclosed are glycan compounds and their use for treating or inhibiting the development of a viral infection, in particular a coronavirus infection, such as a SARS-CoV-2 infection, in a subject, or for treating a condition associated with a viral infection, such as an acute inflammatory condition, cytokine release syndrome (CRS) or cytokine storm, severe acute respiratory syndrome (SARS) or acute respiratory distress syndrome (ARDS).

Description

[0001] This application claims priority to Australian Provisional Patent Application No. 2023902113, filed on June 30, 2023, entitled “Compounds and Uses Thereof,” the contents of which are incorporated herein by reference in their entirety.

[0002] Invention Field

[0003] This invention generally relates to the use of glycanic compounds and their use in treating or inhibiting the development of viral infections (particularly coronavirus infections, such as SARS-CoV-2 infection) in subjects, or in treating conditions associated with viral infections (such as acute inflammatory conditions, cytokine release syndrome (CRS) or cytokine storm, severe acute respiratory syndrome (SARS) or acute respiratory distress syndrome (ARDS)). Background of the Invention

[0005] References to any prior publications (or information obtained from them) or any known matters in this specification do not constitute and should not be regarded as an acknowledgment or endorsement or, in any way, imply that such prior publications (or information obtained from them) or known matters form part of the ordinary general knowledge in the field covered by this specification.

[0006] Viral infections impose a tremendous economic and social burden on society, as evidenced by the recent pandemic caused by the coronavirus SARS-CoV-2. SARS-CoV-2 is a newly emerging virus that causes a frequently fatal respiratory illness, COVID-19. The current pandemic caused by SARS-CoV-2 is a health emergency, necessitating the development of new vaccines and treatments to prevent or treat infections caused by this virus.

[0007] SARS-CoV-2 is closely related to severe acute respiratory syndrome coronavirus (SARS-CoV) (Lu et al., Lancet 2020, 395 (10224), 565-574), and recent studies have confirmed that the SARS-CoV-2 spike protein, like SARS-CoV, uses angiotensin-converting enzyme 2 (ACE2) as a cell receptor to initiate membrane fusion and infection (Hoffmann et al., Cell 2020 181(2): 271-280). SARS-CoV-2 has a higher binding affinity to the ACE2 receptor than SARS-CoV (Wrapp et al., Science 2020, 367(6483), 1260-1263). Many other viruses use glycoproteins on their surface as fusion proteins to interact with and enter host cells to cause infection, including enveloped viruses such as influenza and human immunodeficiency virus (HIV). Therefore, the interaction between viral fusion proteins and host cell ligands is an attractive therapeutic target for many important viral infections.

[0008] Glycosaminoglycans (GAGs) are structurally diverse linear polysaccharides. They occur naturally as glycan portions of proteoglycans and play crucial roles in many biological processes in the human body, including the binding and presentation of certain cytokines / growth factors to their cell surface receptors, thereby enhancing receptor signaling. Glycosaminoglycans in the extracellular matrix also isolate certain cytokines, growth factors, chemokines, and enzymes at specific locations within tissues. However, interactions between GAGs and specific cytokines, growth factors, chemokines, and enzymes can promote physiologically detrimental processes, such as excessive inflammation, cancer growth and metastasis, and various immune responses, leading to counterproductive results. Currently, some of the most well-known GAGs belong to the sulfated polysaccharides of the heparin / heparin sulfate family. Heparin is best known for its anticoagulant activity, but recent data suggest that it can also possess anti-inflammatory activity. Mechanistically, these activities result from certain structures in heparin / heparin sulfate that bind to three-dimensional motifs on proteins. Due to their ability to bind to and modulate the activity of biological modulators, GAGs are considered potential therapeutic agents.

[0009] However, due to their extreme structural heterogeneity, particularly their complex sulfation patterns, and the difficulty in isolating specific GAG structural entities, natural GAGs are not ideal therapeutic agents. This means that, under normal circumstances, the optimal natural GAG structural motif for binding specific cytokines, growth factors, chemokines, or enzymes is unknown. The synthesis of GAG structures is also extremely complex, and even the synthesis of small GAG structures requires numerous synthetic steps. Because the complex sulfation patterns of GAGs contribute to their protein-binding characteristics, multiple synthetic steps are unavoidable when synthesizing GAGs for therapeutic purposes. Therefore, there is a need for GAGs or their analogues or mimics with more suitable therapeutic properties. Invention Overview

[0011] WO 2018 / 068090 A1 discloses a series of compounds with improved synthetic routes compared to GAG, and therefore more suitable therapeutic properties. This invention is based in part on the discovery of specific compounds that bind to viral fusion proteins (e.g., spike proteins) and inhibit the interaction between the fusion proteins and host cells. Therefore, the inventors believe that such compounds can be used to treat and inhibit the development of viral infections in subjects, particularly infections of viruses expressing viral fusion proteins, such as enveloped viral infections (e.g., coronavirus infections, such as SARS-CoV-2 infection), or to treat conditions associated with viral infections (e.g., coronavirus infections), such as acute inflammatory conditions, CRS or cytokine storms, SARS, or ARDS.

[0012] In one respect, compounds of formula I, II, or III are provided:

[0013]

[0014] (I)

[0015]

[0016] (II)

[0017]

[0018] (III)

[0019] Or its pharmaceutically acceptable salt, solvate, or prodrug,

[0020] in:

[0021] L 1 L 3 L 4 L 6 L 7 L 8 L 9 L 10 L 11 L 12 L 13 L 15 L 16 L 17 L 18 and L 19 Independently selected from optionally substituted C1-C5 alkylene groups, optionally substituted C2-C5 alkenyl groups, and optionally substituted C2-C5 ynylene groups;

[0022] L 2 L 5 and L 14Independently selected from optionally substituted C2-C8 alkylene groups, optionally substituted C2-C8 alkenyl groups, and optionally substituted C2-C8 ynylene groups;

[0023] X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 and X 11 Independently selected from CH and N; and

[0024] R 1 Selected from C2-C which are optionally replaced 12 Alkyl groups, optionally substituted C2-C 12 Alkenyl and optionally substituted C2-C 12 Alkyne group.

[0025] In some implementations, X 1 -X 11 It is CH.

[0026] In some implementations, L 1 L 3 L 4 L 6 L 7 L 8 L 9 L 10 L 11 L 12 L 13 L 15 L 16 L 17 L 18 and L 19 Independently selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 ynynyl groups. In a particular embodiment, L 1 L 3 L 4 L 6 L 7 L 8 L 9 L 10 L 11 L 12 L 13 L 15 L 16 L 17 L18 and L 19 It is an optional substituted C1-C3 alkylene group; in particular, a methylene group.

[0027] In some implementations, L 2 Selected from optionally substituted C2-C5 alkylene groups, optionally substituted C2-C5 alkenyl groups, and optionally substituted C2-C5 ynylene groups. In a particular embodiment, L 2 It is an optional substituted C2-C5 alkylene group, such as propylene (e.g., n-propylene).

[0028] In other implementation schemes, L 5 Selected from optionally substituted C3-C6 alkylene groups, optionally substituted C3-C6 alkenyl groups, and optionally substituted C6-C6 ynynyl groups. In a specific embodiment, L 5 It is an optional substituted C3-C6 alkylene group, particularly butylene (e.g., n-butylene).

[0029] In some implementations, L 14 Selected from optionally substituted C2-C4 alkylene groups, optionally substituted C2-C4 alkenyl groups, and optionally substituted C2-C4 ynylene groups. In a particular embodiment, L 14 It is a C2-C4 alkylene group that is optionally substituted, particularly an ethylene group.

[0030] In the specific implementation plan, R 1 Selected from C6-C which are optionally replaced 10 Alkyl groups, optionally substituted C6-C 10 Alkenyl and optionally substituted C6-C 10 Alkyne group. In a particular embodiment, R 1 C6-C is arbitrarily replaced. 10 Alkyl groups, such as octyl groups.

[0031] In some embodiments, the compound is a compound of formula IV:

[0032]

[0033] (IV)

[0034] Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0035] In an alternative embodiment, the compound is a compound of formula V:

[0036]

[0037] (V)

[0038] Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0039] In other embodiments, the compound is a compound of formula VI:

[0040]

[0041] (VI)

[0042] Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0043] In a particular embodiment, the compound is in the form of a salt, specifically a sodium salt.

[0044] In another aspect, a pharmaceutical composition is provided comprising a compound of formula I, II, III, IV, V or VI, or a pharmaceutically acceptable salt, solvate or prodrug thereof, and a pharmaceutically acceptable carrier or diluent.

[0045] Further, compounds of formula I, II, III, IV, V or VI, or pharmaceutically acceptable salts, solvates or prodrugs thereof, are provided for use in therapy.

[0046] In another aspect, a method is provided for treating or inhibiting the development of a viral infection in a subject, the method comprising, consisting of, or substantially consisting of: administering to the subject a compound of formula I, II, III, IV, V, VI, or VII, or a pharmaceutically acceptable salt, solvate, or prodrug thereof:

[0047]

[0048] (VII)

[0049] In certain embodiments, the infection is caused by a virus expressing a viral fusion protein (such as an enveloped virus). In some embodiments, the infection is caused by a virus interacting with cells expressing the ACE2 peptide. While any viral infection is considered, in certain embodiments, the viral infection is a coronavirus infection, specifically one in which the coronavirus is capable of causing SARS. In some embodiments, the coronavirus is a beta-coronavirus, such as a beta-coronavirus selected from lineage A, lineage B, lineage C, and lineage D. In some embodiments, the beta-coronavirus is lineage B beta-coronavirus, such as SARS-CoV or SARS-CoV-2; particularly SARS-CoV-2. In alternative embodiments, the beta-coronavirus is lineage C beta-coronavirus, such as MERS-CoV.

[0050] In some embodiments, the compound is administered orally, by inhalation, intranasally, topically, or intravenously.

[0051] While the application of a single compound is considered, the present invention also considers the application of a second compound of formula I, II, III, IV, V, VI, or VII, or a pharmaceutically acceptable salt, solvate, or prodrug thereof. In some embodiments, the method comprises the application of a compound of formula V, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, and a compound of formula VI, or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

[0052] In another aspect, a method for inhibiting the interaction between a virus and cells expressing the ACE2 polypeptide is provided, the method comprising, consisting of, or substantially consisting of contacting the virus with a compound of formula I, II, III, IV, V, VI, or VII, or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

[0053] In some implementations, the interaction is one or both of virus binding to a cell and virus entry into a cell.

[0054] In some embodiments, the cells are lung cells (e.g., alveolar cells), intestinal cells, endothelial cells, epithelial cells (e.g., nasal or nasopharyngeal epithelial cells), kidney cells (e.g., brush border cells of proximal renal tubular epithelial cells), or arterial smooth muscle cells. In a particular embodiment, the cells are respiratory tract cells.

[0055] In certain embodiments, the virus is a coronavirus, particularly one capable of causing SARS. In some embodiments, the coronavirus is a beta-coronavirus, such as one selected from lineage A, lineage B, lineage C, and lineage D. In some embodiments, the beta-coronavirus is lineage B, such as SARS-CoV or SARS-CoV-2; particularly SARS-CoV-2. In alternative embodiments, the beta-coronavirus is lineage C, such as MERS-CoV.

[0056] In some embodiments, the compound is administered orally, by inhalation, intranasally, topically, or intravenously.

[0057] In some embodiments, the method further comprises administering a second compound of formula I, II, III, IV, V, VI, or VII, or a pharmaceutically acceptable salt, solvate, or prodrug thereof. Exemplary embodiments comprise administering a compound of formula V, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, and a compound of formula VI, or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

[0058] In another aspect, a method is provided for treating an acute inflammatory condition in a subject, wherein the condition is associated with a coronavirus infection, the method comprising, consisting of, or substantially consisting of administering to the subject a compound of formula I, II, III, IV, V, VI, or VII, or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

[0059] In some embodiments, the coronavirus is a beta-coronavirus, such as lineage A beta-coronavirus, lineage B beta-coronavirus, lineage C beta-coronavirus, or lineage D beta-coronavirus. In some embodiments, the beta-coronavirus is lineage B beta-coronavirus, such as SARS-CoV or SARS-CoV-2; particularly SARS-CoV-2. In alternative embodiments, the beta-coronavirus is lineage C beta-coronavirus, such as MERS-CoV.

[0060] In some embodiments, the acute inflammatory condition is associated with the presence of CRS or a cytokine storm, specifically wherein the CRS or cytokine storm comprises an increase of at least 50% compared to the baseline state of one or more cytokines or chemokines selected from IFN-γ, IFN-β, TNF-α, IL-1β, IL-6, IL-17A, CCL3, CXCL2, and CXCL8.

[0061] In some embodiments, the subject has CRS and exhibits one or more symptoms selected from the following: fever, fatigue, loss of appetite, muscle and joint pain, nausea, vomiting, diarrhea, rash, rapid breathing, rapid heartbeat, hypotension, seizures, headache, confusion, delirium, hallucinations, tremor, and coordination disorders. In alternative embodiments, the subject has a cytokine storm and exhibits one or more symptoms selected from the following: high fever, swelling and redness, extreme fatigue, nausea, bleeding, coagulation, visceral organ damage, and shock, or any combination thereof.

[0062] In some implementations, the acute inflammatory condition is associated with multisystem inflammatory syndrome in children (MIS-C), for example, wherein the subject has one or more symptoms selected from the following: fever, vomiting, diarrhea, stomach pain, rash, red eyes, red or swollen lips and tongue, unusual fatigue, red or swollen hands or feet, severe stomach pain, cardiac symptoms (including chest pain, palpitations, and shortness of breath), cyanosis of lips or face, confusion, inability to be awakened or remain conscious, abdominal pain with vomiting and diarrhea, rash and swelling of the extremities, syncope, and new-onset hypotension.

[0063] The association between acute inflammatory symptoms and systemic inflammatory response syndromes (SIRS) is also considered. In some implementations, the subject may have stage 1 SIRS (local response), stage 2 SIRS (early compensatory anti-inflammatory response syndrome (CARS), designed to maintain immune homeostasis), stage 3 SIRS (pro-inflammatory SIRS, which leads to progressive endothelial dysfunction, coagulopathy, and activation of coagulation pathways), stage 4 SIRS (characterized by CARS replacing SIRS, which leads to a state of relative immunosuppression. Therefore, individuals are prone to secondary or nosocomial infections, thus perpetuating the sepsis cascade), or stage 5 SIRS (manifested as multiple organ dysfunction syndrome (MODS) with persistent dysregulation of SIRS and CARS responses).

[0064] In some embodiments, the acute inflammatory condition is associated with acute respiratory distress syndrome (ARDS). In such embodiments, the subject may have one or more symptoms selected from the following: mild, moderate, or severe hypoxemia (determined by arterial oxygen partial pressure / inspiratory oxygen concentration (PaO2 / FiO2) or positive end-expiratory pressure (PEEP), bilateral lung shadows, respiratory failure, shortness of breath, dyspnea, cough, fever, tachycardia, hypotension, confusion, extreme fatigue, rapid breathing, organ failure, chest pain, cyanosis of nails or lips, changes in the levels of one or more inflammatory markers, or the need for mechanical ventilation.

[0065] In some implementations, the acute inflammatory symptoms are associated with SARS. The subject may have, for example, one or more of the following symptoms: acute febrile illness, weakness, fatigue, headache, flushing, diarrhea, nausea, vomiting, cough (including dry cough), sore throat, runny nose, nasal congestion, production of pro-inflammatory mediators, vascular leakage, and organ failure.

[0066] The present invention further provides a method for treating CRS or cytokine storm in a subject, wherein the CRS or cytokine storm is associated with coronavirus infection, the method comprising, consisting of, or substantially consisting of administering to the subject a compound of formula I, II, III, IV, V, VI, or VII or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

[0067] In some embodiments, the coronavirus is a beta-coronavirus, such as lineage A beta-coronavirus, lineage B beta-coronavirus, lineage C beta-coronavirus, or lineage D beta-coronavirus. In some embodiments, the beta-coronavirus is lineage B beta-coronavirus, such as SARS-CoV or SARS-CoV-2; particularly SARS-CoV-2. In alternative embodiments, the beta-coronavirus is lineage C beta-coronavirus, such as MERS-CoV.

[0068] In a particular embodiment, the CRS or cytokine storm comprises an increase of at least 50% compared to the baseline state of one or more cytokines or chemokines selected from IFN-γ, IFN-β, TNF-α, IL-1β, IL-6, IL-17A, CCL3, CXCL2, and CXCL8.

[0069] In some implementations, the subject has CRS and has one or more symptoms selected from the following: fever, fatigue, loss of appetite, muscle and joint pain, nausea, vomiting, diarrhea, rash, rapid breathing, rapid heartbeat, low blood pressure, seizures, headache, confusion, delirium, hallucinations, tremor, and coordination disorder.

[0070] In some implementations, the subject has a cytokine storm and has one or more symptoms selected from the following: high fever, swelling and redness, extreme fatigue, nausea, bleeding, coagulation, visceral organ damage and shock, or any combination thereof.

[0071] The CRS or cytokine storm can be associated with MIS-C. In such an implementation, the subject may have one or more symptoms selected from the following: fever, vomiting, diarrhea, stomach pain, rash, red eyes, red or swollen lips and tongue, unusual fatigue, red or swollen hands or feet, severe stomach pain, cardiac symptoms (including chest pain, palpitations, and shortness of breath), cyanosis of lips or face, confusion, inability to be awakened or remain conscious, abdominal pain with vomiting and diarrhea, rash and swelling of the extremities, syncope, and new-onset hypotension.

[0072] In some implementations, the CRS or cytokine storm is associated with SIRS, such as stage 1 SIRS, stage 2 SIRS, stage 3 SIRS, stage 4 SIRS, or stage 5 SIRS.

[0073] In some embodiments, the CRS or cytokine storm is associated with ARDS. In these embodiments, the subject may have one or more symptoms selected from the following: mild, moderate, or severe hypoxemia (determined by arterial oxygen partial pressure / inspiratory oxygen concentration (PaO2 / FiO2) or positive end-expiratory pressure (PEEP), bilateral lung shadows, respiratory failure, shortness of breath, dyspnea, cough, fever, tachycardia, hypotension, confusion, extreme fatigue, rapid breathing, organ failure, chest pain, cyanosis of nails or lips, changes in the levels of one or more inflammatory markers, or the need for mechanical ventilation.

[0074] In a particular implementation, the CRS or cytokine storm is associated with SARS. For example, the subject may have one or more symptoms selected from the following: acute febrile illness, weakness, fatigue, headache, flushing, diarrhea, nausea, vomiting, cough (including dry cough), sore throat, runny nose, nasal congestion, production of pro-inflammatory mediators, vascular leakage, and organ failure.

[0075] Another aspect of the invention provides a method for treating SARS in a subject, wherein the SARS is associated with coronavirus infection, the method comprising, consisting of, or substantially consisting of administering to the subject a compound of formula I, II, III, IV, V, VI, or VII or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

[0076] In some embodiments, the coronavirus is a beta-coronavirus, such as lineage A beta-coronavirus, lineage B beta-coronavirus, lineage C beta-coronavirus, or lineage D beta-coronavirus. In some embodiments, the beta-coronavirus is lineage B beta-coronavirus, such as SARS-CoV or SARS-CoV-2; particularly SARS-CoV-2. In alternative embodiments, the beta-coronavirus is lineage C beta-coronavirus, such as MERS-CoV.

[0077] The subject may have one or more of the following symptoms: acute febrile illness, weakness, fatigue, headache, flushing, diarrhea, nausea, vomiting, cough (including dry cough), sore throat, runny nose, nasal congestion, production of pro-inflammatory mediators, vascular leakage and organ failure or any combination thereof.

[0078] In some embodiments, the SARS is associated with the presence of CRS or a cytokine storm, for example, wherein the CRS or cytokine storm comprises an increase of at least 50% compared to the baseline state of one or more cytokines or chemokines selected from IFN-γ, IFN-β, TNF-α, IL-1β, IL-6, IL-17A, CCL3, CXCL2, and CXCL8.

[0079] In some implementations, the SARS is associated with MISC-C. In such implementations, the subject has one or more symptoms selected from the following: fever, vomiting, diarrhea, stomach pain, rash, red eyes, red or swollen lips and tongue, unusual fatigue, red or swollen hands or feet, severe stomach pain, cardiac symptoms (including chest pain, palpitations, and shortness of breath), cyanosis of the lips or face, confusion, inability to be awakened or remain conscious, abdominal pain with vomiting and diarrhea, rash and swelling of the extremities, syncope, and new-onset hypotension.

[0080] The SARS mentioned can also be associated with SIRS. For example, the subjects may have stage 1 SIRS, stage 2 SIRS, stage 3 SIRS, stage 4 SIRS, or stage 5 SIRS.

[0081] This document further provides the use of compounds of formula I, II, III, IV, V, VI, or VII, or pharmaceutically acceptable salts, solvates, or prodrugs thereof, in the preparation of a medicament for the treatment or inhibition of the development of a viral infection in a subject.

[0082] Another aspect provides the use of compounds of formula I, II, III, IV, V, VI, or VII, or pharmaceutically acceptable salts, solvates, or prodrugs thereof, in the preparation of a medicament for treating an acute inflammatory condition in a subject, said condition being associated with coronavirus infection.

[0083] Further consideration is given to the use of compounds of formula I, II, III, IV, V, VI, or VII, or pharmaceutically acceptable salts, solvates, or prodrugs thereof, in the preparation of a drug for treating CRS or cytokine storm in a subject, wherein the CRS or cytokine storm is associated with coronavirus infection.

[0084] The present invention further provides the use of compounds of formula I, II, III, IV, V, VI or VII or pharmaceutically acceptable salts, solvates or prodrugs thereof in the preparation of a medicament for treating SARS in a subject, wherein the SARS is associated with coronavirus infection.

[0085] Another aspect provides compounds of formula I, II, III, IV, V, VI, or VII, or their pharmaceutically acceptable salts, solvates, or prodrugs, for the treatment or inhibition of the development of a viral infection in a subject.

[0086] In another aspect, compounds of formula I, II, III, IV, V, VI, or VII, or pharmaceutically acceptable salts, solvates, or prodrugs thereof, are provided for treating acute inflammatory conditions in subjects associated with coronavirus infection.

[0087] This article also considers compounds of formula I, II, III, IV, V, VI, or VII, or their pharmaceutically acceptable salts, solvates, or prodrugs, for use in treating subjects with CRS or cytokine storms associated with coronavirus infection.

[0088] In another aspect, compounds of formula I, II, III, IV, V, VI, or VII, or pharmaceutically acceptable salts, solvates, or prodrugs thereof, are provided for the treatment of SARS in subjects, wherein the SARS is associated with coronavirus infection. Brief description of the attached diagram

[0090] Figure 1 The images show the binding of the SARS-CoV-2 spike protein receptor-binding domain (RBD) to Calu-3 cells expressing ACE2 in the presence of G24, G22, G17, or heparin, compared to cells in the absence of the test compounds. The presence of the RBD on the cells was detected using anti-His antibody (red).

[0091] Figure 2 The graph shows the percentage reduction in lesions in Vero E6 monkey kidney cells in the presence of SARS-CoV-2 virus (Australia / VIC01 / 2020) and in the presence and absence of different concentrations of the test compounds (G4, G17, G19, G22, or G24). Ungraded heparin (UFH) was used as a control compound. This data is from a single first run (Run 1).

[0092] Figure 3 The graph shows the percentage reduction in lesions in Vero E6 monkey kidney cells in the presence of SARS-CoV-2 virus (Australia / VIC01 / 2020) and in the presence and absence of different concentrations of the test compounds (G4, G17, G19, G22, or G24). This data is from a single second run (Run 2).

[0093] Figure 4 The figure illustrates the inhibition of IL-8 (CXCL8)-dependent chemotaxis in human promyelocytic HL-60 cells treated with dimethyl sulfoxide (DMSO) in the presence of the test compounds (G19, G22, and G24) at a concentration of 50 μg / mL. Heparin was used as a control compound. Mean, upper limit, and lower limit values ​​are shown. Invention Details

[0095] 1. Definition

[0096] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. While any methods and materials similar to or equivalent to those described herein may be used to practice or test the invention, preferred methods and materials are described. For the purposes of this invention, the following terms are defined below.

[0097] The articles “a” and “a kind” are used in this text to indicate one or more (i.e., at least one) grammatical object of the article. As an example, “a component” refers to one or more components.

[0098] "Approximately" means a quantity, level, value, number, frequency, percentage, size, size, quantity, weight, or length that varies by as much as 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% compared to a reference quantity, level, value, number, frequency, percentage, size, quantity, weight, or length.

[0099] As used herein, the term "acute inflammatory condition" refers to a condition in which acute inflammation is present and exhibits a rapid, transient (minutes to days), relatively homogeneous response to acute injury, characterized by the accumulation of fluids, plasma proteins, and neutrophils. In acute inflammation, the removal of the stimulus prevents the recruitment of monocytes (which, upon appropriate activation, transform into macrophages) to the inflamed tissue, and existing macrophages leave the tissue via the lymphatic system. Examples of harmful factors that cause acute inflammation include, but are not limited to, pathogens (e.g., bacteria, viruses, parasites), foreign bodies of exogenous (e.g., asbestos) or endogenous (e.g., urate crystals, immune complexes), and physical (e.g., burns) or chemical (e.g., corrosive) factors. Typically, the physiological changes accompanying acute inflammation encompass four main characteristics: (1) vasodilation, which results in a net increase in blood flow and is one of the earliest physical responses to acute tissue injury; (2) in response to inflammatory stimuli, the endothelial cells lining the venules contract, and intracellular junctions expand to create gaps, leading to increased vascular permeability, which allows plasma proteins and blood cells to leak from the vessels; (3) inflammation is usually characterized by a strong infiltration of leukocytes, particularly neutrophils (polymorphonuclear cells), at the site of inflammation. These cells promote tissue damage by releasing toxic substances at the vessel walls or in undamaged tissues; and (4) fever, generated by pyrogens released by leukocytes in response to specific stimuli. Other symptoms include decreased sense of smell and adverse effects on the cardiovascular system.

[0100] As used in this article, the term "acute respiratory distress syndrome" or "ARDS" refers to a life-threatening lung condition that prevents enough oxygen from reaching the lungs and entering the bloodstream. ARDS is also known as noncardiac pulmonary edema, osmotic pulmonary edema, stiff lung, shock lung, or acute lung injury. ARDS can be caused by any serious lung injury. Some common causes include, but are not limited to: aspiration of vomit into the lungs (aspiration), inhalation of chemicals, lung transplantation, pneumonia, septic shock (systemic infection), and trauma.

[0101] The terms "parallel administration," "concurrent administration," or "co-administration," etc., refer to the administration of a single composition containing two or more agents, or the simultaneous or sequential administration of each agent as a separate composition and / or via a separate route, within a sufficiently short period of time, to achieve the same effect as when all such agents are administered as a single composition. "Simultaneously" means that the agents are administered at substantially the same time and ideally together in the same composition. "Simultaneously" means that the agents are administered close together in time, for example, the administration of one agent is about 1 minute to about 1 day apart from the administration of another agent. Any period of time is useful. However, in general, when not administered simultaneously, the agents are administered from about 1 minute to about 8 hours, and appropriately within less than about 1 hour to about 4 hours. When administered simultaneously, the agents are appropriately administered to the same site on the subject. The term "same site" includes an exact location but can be within about 0.5 to about 15 cm, preferably within about 0.5 to about 5 cm. As used herein, the term "separately" means applying the agent at intervals, such as from approximately one day to several weeks or months. The agent may be applied in any order. As used herein, the term "in sequence" means applying the agent sequentially, such as at intervals of minutes, hours, days, or weeks. If appropriate, the agent may be applied in a regular, repeated cycle.

[0102] The term "pharmaceutical" includes compounds that induce desired pharmacological and / or physiological effects. This term also encompasses pharmaceutically acceptable and pharmacologically active components of those compounds specifically mentioned herein, including, but not limited to, salts, esters, amides, prodrugs, active metabolites, analogs, etc. When used with the above term, it should be understood that this includes the active agent itself, as well as pharmaceutically acceptable and pharmacologically active salts, esters, amides, prodrugs, metabolites, analogs, etc. The term "pharmaceutical" should not be interpreted narrowly, but rather extended to small molecules, protein molecules (such as peptides, polypeptides, and proteins) and compositions comprising them, genetic molecules (such as RNA, DNA, and their analogues and chemical analogs), and cellular preparations.

[0103] The term "alkyl" refers to a straight-chain or branched aliphatic hydrocarbon group, including C1-C64 groups. 12 Alkyl, C2-C 12 Alkyl, C6-C 10 Alkyl, C7-C9 alkyl, and C8 alkyl (octyl), unless otherwise specified. In certain embodiments, the alkyl group is a straight-chain aliphatic hydrocarbon group, such as methyl, ethyl, n-propyl, n-butyl, pentyl, hexyl, heptyl, octyl, etc.

[0104] As used herein, the term "alkylene" refers to a divalent alkyl group having 1 to 8 carbon atoms, including 2 to 8 carbon atoms, 3 to 6 carbon atoms, 1 to 5 carbon atoms, 1 to 3 carbon atoms, 2 to 5 carbon atoms, and 2 to 4 carbon atoms. Examples of such alkylene groups include methylene (-CH2-), ethylene (-CH2CH2-), propylene including n-propylene (-CH2CH2CH2-), butylene including n-butylene (-CH2CH2CH2CH2-), etc.

[0105] The term "alkenyl" refers to an aliphatic hydrocarbon group containing at least one carbon-carbon double bond, and it can be straight-chain or branched (preferably straight-chain), including C2-C... 12 alkenyl, C6-C 10 Alkenyl, C7-C9 alkenyl, and C8 alkenyl, unless otherwise specified. The group may contain multiple double bonds in the positive chain, and each double bond may be oriented independently as either E or Z. Exemplary alkenyl groups include, but are not limited to, vinyl, propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl, and nonenyl.

[0106] As used herein, the term "alkenyl" refers to a divalent alkenyl group having 2 to 8 carbon atoms, including 3 to 6 carbon atoms, 2 to 5 carbon atoms, 2 to 4 carbon atoms, and 2 to 3 carbon atoms.

[0107] "Alkyne" refers to an aliphatic hydrocarbon group containing at least one carbon-carbon triple bond (e.g., one carbon-carbon triple bond), and it can be straight-chain or branched, preferably straight-chain, including C2-C 12 alkynyl group, C6-C 10 Alkynyl, C7-C9 alkynyl and C8 alkynyl, unless otherwise specified.

[0108] The term "ethynyl" refers to a divalent ethynyl group having 2 to 8 carbon atoms, including 3 to 6 carbon atoms, 2 to 5 carbon atoms, 2 to 4 carbon atoms, and 2 to 3 carbon atoms.

[0109] The term “and / or” as used in this document means and covers any and all possible combinations of one or more of the related listed items, as well as cases where no combination is possible when interpreted in an alternative manner (or).

[0110] As used herein, the term "antimicrobial agent" means any agent with antimicrobial activity, i.e., capable of inhibiting or reducing the growth of microorganisms and / or killing microorganisms, for example, by at least about 30%, at least about 40%, at least about 50%, at least about 75%, at least about 90%, or more, compared to the absence of an antimicrobial agent. The term "antimicrobial agent" encompasses agents that inhibit or reduce the growth of microorganisms and / or kill microorganisms through direct interaction with the cells of the microorganisms and / or the host cells in which the microorganisms reside or are located. Non-limiting examples of antimicrobial agents include silver nanoparticles, small molecules, peptides, peptide mimics, antibodies or fragments thereof, nucleic acids, enzymes (e.g., antimicrobial metalloendopeptides such as lysostaphin), aptamers, drugs, antibiotics, chemicals, or any entity that can inhibit the growth of microorganisms and / or kill microorganisms. Examples of antimicrobial peptides that may be included in the compositions described herein include, but are not limited to: mefloquine, strychnine A, antimycin, thiazolinone, strychnine, diuron, iodoacetamide, potassium tellurite hydrate, aDL-vinylglycine, N-ethylmaleimide, L-allylglycine, diarylquinoline, betaine aldehyde chloride, acivcin, pscifuraine, sulfoxide imide, diaminopimelic acid, 4-phospho-D-erythronhydroxamic acid, motexafen gadolinium and / or xycitrin or modified forms thereof or the like. Representative antimicrobial agents include antibiotics, antifungal agents, antiprotozoal agents, antimalarial agents, antituberculosis agents, and antiviral agents, and any mixtures thereof. In some embodiments, the antimicrobial agent is an antiviral agent, which encompasses agents that effectively inhibit viral formation and / or replication in mammals. This includes agents that interfere with host or viral mechanisms essential for viral formation and / or replication in mammals. Such agents may be selected from immunomodulators, inhibitors of viral polymerases, or inhibitors of other targets in the viral life cycle.Examples of antiviral agents include α-methyl-1-amantadine, hydroxyethoxymethylguanine, amantadine, 5-iodine-2'-deoxyuridine, trifluorothymidine, AZT, cytarabine adenine, abacavir, acyclovir, acyclovir, adefovir, amantadine, ampravir, ampligen, arbidol, atazanavir, atripla, boprevir, cidofovir, dalunavir, duravir, darunavir, deraviridine, doxorubicin, doxorubicin, edaurenin, efavirenz, emtricitabine, enfuviride, entecavir, Entry, inhibitors, famciclovir, fomivir, furazonavir, phosphonoformic acid, phosphonoacetic acid, ganciclovir, ibatabin, imunovir, idoxuridine, imiquimod, indinavir, inosine, integrase inhibitors, and interferon III. Type I interferon, type II interferon, type I interferon, interferon, lamivudine, lopinavir, cyclovir, maraviro, morpholine guanidine, meticazone, nelfinavir, nevirapine, nexavir, nucleoside analogs, oseltamivir, (Tamiflu), pegylated interferon alpha-2a, penciclovir, pelamivir, proconazole, podophyllotoxin, raltegravir, ribavirin, amantadine, ritonavir, pyramidine, saquinavir, stavudine, tenofovir, tenofovir, disoproxil, lipranavir, trifluralin, trifluralin, triamcinolone, suvastatin, valacyclovir (Valtrex), valacyclovir, velivirol, vidarabine, taribivudine hydrochloride, zalcitabine, zanamivir (Relenza), and zidovudine.

[0111] The terms “angiotensin-converting enzyme 2 polypeptide,” “ACE2 polypeptide,” and “ACE2” are used interchangeably herein to refer to a polypeptide comprising the amino acid sequence of a precursor ACE2 polypeptide (e.g., the human precursor ACE2 polypeptide listed in GenBank Accession NP_001358344) or its processed form (e.g., a mature ACE2 polypeptide with the signal peptide (e.g., defined by amino acids 1-17) and / or with the carboxyl-terminal portion (e.g., defined by amino acids 709-805) removed), or a biologically active fragment of a precursor or processed form of the ACE2 polypeptide, and allelic variants of any of these. The degree and location of glycosylation or other post-translational modifications may vary depending on the nature of the chosen host and host cellular environment.

[0112] As used herein, the term "cell expressing ACE2 peptide" refers to a cell that typically expresses the ACE2 peptide on its surface. Representative cells expressing the ACE2 peptide include myeloid cells of the lung, arteries, heart, kidney, and intestine. Cells expressing ACE2 are suitably selected from lung cells (e.g., alveolar cells), intestinal cells, endothelial cells, and arterial smooth muscle cells. In a specific embodiment, the ACE2-expressing cells are respiratory tract cells (e.g., lung cells such as alveolar cells).

[0113] As used herein, the term "antagonist" and its grammatical equivalents refer to a molecule that partially or completely inhibits the action of another molecule (such as a receptor or intracellular mediator) through any mechanism. In the context of this invention, the term "antagonist" refers to a compound that partially or completely inhibits the interaction or formation of a complex between a fusion protein (e.g., the spike protein) and a host cell ligand (such as a host cell surface protein (e.g., CE2)). In specific embodiments, the antagonist is a direct antagonist that binds to or otherwise interacts with a viral fusion protein (e.g., the spike protein).

[0114] When used to describe a condition associated with a viral infection (such as coronavirus infection), the term "associated with" means that the viral infection directly or indirectly contributes to the onset or progression of the condition, including one or more symptoms of the condition. For example, the viral infection may directly cause the onset (i.e., development) of the condition or the development of one or more symptoms of the condition. Alternatively or additionally, the viral infection may cause the progression (i.e., worsening) of the condition or one or more symptoms of the condition. Similarly, a condition associated with the presence of CRS or a cytokine storm, MIS-C, ARDS, SARS, or SIRS is a condition in which the presence of CRS or a cytokine storm, MIS-C, ARDS, SARS, or SIRS contributes to the onset or progression of the condition (including one or more symptoms of the condition).

[0115] Throughout this specification and the following claims, unless the context otherwise requires, the word “comprising” and variations such as “including” and “containing” should be understood to imply inclusion of the stated integers or steps or groups of integers or steps, but not to exclude any other integers or steps or groups of integers or steps. Thus, the use of the term “comprising” indicates that the listed integers are necessary or mandatory, but other integers are optional and may be present or absent. “Constitutes of” is intended to include and limit to what follows the phrase “consisting of”. Thus, the phrase “consisting of” indicates that the listed elements are necessary or mandatory, and no other elements may be present. “Substantially constitutes of” means to include any element listed following the phrase, and is limited to other elements that do not interfere with or contribute to the activity or function prescribed for the listed elements. Thus, the phrase “substantially constitutes of” indicates that the listed elements are necessary or mandatory, but other elements are optional and may be present or absent depending on whether they affect the activity or function of the listed elements.

[0116] The term "cytokine release syndrome" or "CRS" refers to a systemic inflammatory response syndrome (SIRS) that can be triggered by a variety of factors, such as infection and certain medications. It stands for cytokine storm syndrome (CSS) and occurs when a large number of white blood cells are activated and release inflammatory cytokines, which in turn activate even more white blood cells. CRS is also an adverse reaction to certain monoclonal antibody drugs and adoptive T-cell therapy. When caused by medication, it is also called an infusion reaction. The terms cytokine storm and CRS are often used interchangeably, but despite their similar clinical phenotypes, they differ in characteristics. When treatment-induced, CRS symptoms may be delayed until days or weeks after treatment. Immediate-onset CRS is a cytokine storm, although severe cases of CRS are also referred to as cytokine storms.

[0117] As used in this article, the term "cytokine storm" refers to an overactivated cytokine cascade or hypercytokineemia, characterized by the excessive or uncontrolled release of pro-inflammatory cytokines, which can be associated with a variety of infectious and non-infectious diseases or disorders. Cytokine storm syndrome is associated with a group of disorders, such as, but not limited to, influenza, asthma, hantavirus pulmonary syndrome, SIRS, macrophage activation syndrome, SARS, COVID-19, and disseminated intravascular coagulation (DIC), representing a variety of inflammatory causes. Typically, the main symptoms of a cytokine storm are high fever, swelling and redness, extreme fatigue, and nausea. In some cases, the immune response can lead to bleeding, coagulation, visceral organ damage, or shock, and can be fatal.

[0118] As used herein, the term "dosage unit form" refers to a physically discrete unit suitable as a unit dose for use in a subject to be treated, each unit containing a predetermined amount of active substance calculated to produce the desired therapeutic effect, and a pharmaceutically acceptable medium as required.

[0119] In the context of treating or preventing a disease, an "effective amount" refers to a quantity of a drug or composition administered to an individual who requires such treatment or prevention, whether as a single dose or as part of a series, that is effective in preventing the occurrence of symptoms of the disease, controlling such symptoms, and / or treating existing symptoms of the disease. The effective amount will vary depending on the health and physical condition of the individual being treated, the individual's taxonomy, the formulation of the composition, an assessment of the medical condition, and other relevant factors. It is anticipated that the amount will fall within a relatively broad range that can be determined through routine testing. Non-restrictive symptoms of viral infections (e.g., coronavirus infection) include acute febrile illness, weakness, fatigue, headache, flushing, diarrhea, nausea, vomiting, cough (including dry cough), sore throat, runny nose, nasal congestion, and symptoms of systemic inflammatory response syndrome in severe illness, including the production of pro-inflammatory mediators, vascular leakage, and organ failure.

[0120] As used herein, the term "fusion protein" refers to a protein on the surface of a virus that interacts with proteins on the surface of a host cell or on the cell membrane of a host cell, thereby enabling the virus to bind to and / or enter the host cell. Exemplary fusion proteins include the spike protein of coronaviruses.

[0121] As used herein, the phrase "inhibit the development of..." refers to preventive treatment that enhances a subject's resistance to the occurrence of a disease or condition, or in other words, reduces the likelihood of a subject developing a disease or condition; and treatment after a disease or condition has begun, with the aim of reducing or eliminating the disease or condition or preventing its worsening. Within its scope, the phrase also includes preventing the development of the disease or condition in subjects who may be susceptible but have not yet been diagnosed with it.

[0122] As used in this article, the term "inhibitor" refers to an agent that reduces or inhibits at least one function or biological activity of a target molecule, such as a viral fusion protein (e.g., the spike protein).

[0123] When referring to the interaction between two molecules, the term "interaction" (including its grammatical equivalents) means the physical contact between the molecules. Typically, such an interaction results in the activity of one or both of the molecules (which produces biological effects, such as viral binding to and / or viral entry into host cells). This physical contact usually requires the binding or association of molecules with each other and may involve the formation of an induced or paramagnetic magnetic field, covalent bond formation, ionic interactions (such as, for example, interactions occurring in an ionic lattice), hydrogen bonds, or alternatively, van der Waals interactions, such as, for example, dipole-dipole interactions, dipole-induced dipole interactions, induced dipole-induced dipole interactions, or repulsive interactions, or any combination of the foregoing attractive forces.

[0124] The term “multisystem inflammatory syndrome in children” or “MIS-C” as used in this article is a rare, life-threatening condition in which different parts of the body may become inflamed, including the heart, lungs, kidneys, brain, skin, eyes, or gastrointestinal organs. Children with MIS-C may have fever and a variety of symptoms, including abdominal (intestinal) pain, vomiting, diarrhea, neck pain, rash, red eyes, or feeling unusually tired.

[0125] The term "optionally substituted" as used throughout this specification means that the group may be further substituted with one or more non-hydrogen substituents or not substituted. In some embodiments, the substituent is one or more groups independently selected from halogens, C1-C6 alkyl (e.g., methyl or ethyl), C2-C6 alkenyl and C2-C6 ynyl, particularly halogens and C1-C6 alkyl. In some embodiments, the optional substituent is one or more groups independently selected from methyl, ethyl and halogens, particularly methyl. In exemplary embodiments, each optionally substituted group (e.g., alkylene, alkenylene, ynylene, alkyl, alkenyl or ynyl) may be substituted with 1 to 3 substituents, including 1, 2 or 3 substituents, such as 1, 2 or 3 methyl groups.

[0126] The terms “patient,” “subject,” “host,” or “individual,” used interchangeably herein, refer to any subject requiring treatment or prevention, particularly a vertebrate subject, and even more specifically, a mammalian subject. Suitable vertebrates within the scope of this disclosure include, but are not limited to, any member of the subphylum Chordata, including primates (e.g., humans, monkeys, and apes), and including monkey species such as those in the genera *Macaques* (e.g., cynomolgus monkeys, such as tufted monkeys). Macaca fascicularis ) and / or rhesus monkeys ( Macaca mulatta )) and baboons ( Papio ursinus), as well as marmosets (species from the genus Callithrix), squirrel monkeys (species from the genus Saimiri), and little marmosets (species from the genus Saguinus), and ape species such as chimpanzees ( Pan troglodytes Animals included include rodents (e.g., mice, rats, guinea pigs), rabbits (e.g., rabbits, hares), bovids (e.g., cattle), sheep (e.g., sheep), goats (e.g., goats), pigs (e.g., pigs), equines (e.g., horses), canines (e.g., dogs), felines (e.g., cats), birds (e.g., chickens, turkeys, ducks, geese, companion birds such as canaries, lovebirds, etc.), marine mammals (e.g., dolphins, whales), reptiles (e.g., snakes, frogs, lizards, etc.), and fish. In a specific implementation, the subject is a primate (such as a human) that requires treatment or suppression of the development of a viral infection. However, it should be understood that the terms "patient," "subject," "host," or "individual" do not imply the presence of symptoms.

[0127] A "pharmaceutically acceptable carrier" is a drug delivery system composed of a substance that is biologically or otherwise undesirable, meaning that the substance can be administered to a subject in conjunction with a selected active agent without causing any or serious adverse reactions. Carriers may include excipients and other additives such as diluents, fillers, detergents, colorants, wetting agents or emulsifiers, pH buffers, preservatives, etc.

[0128] Similarly, the “pharmacologically acceptable” salts, solvates, esters, or prodrugs of the compounds provided herein are not biologically or otherwise undesirable salts, solvates, esters, or prodrugs.

[0129] When describing the level of a substance and / or phenomenon in a first sample relative to the level in a second sample, the terms “reduction,” “inhibition,” “lowering,” “blocking,” and grammatically equivalent terms mean that the amount of the substance and / or phenomenon in the first sample is statistically significantly lower than the amount in the second sample, as determined using any statistical analysis method accepted in the art. When these terms are used to indicate the effect of a compound or agent, the first sample may be a sample in the presence of the compound or agent, and the second sample may be a control sample without the compound or agent. In one embodiment, reduction can be determined subjectively, for example, when a patient mentions their subjective feelings about disease symptoms, such as pain, headache, fatigue, nausea, motor symptoms, cough, sore throat, nasal congestion, runny nose, etc. In another embodiment, reduction can be determined objectively, for example, when the viral load (e.g., viral load) in a patient's sample is lower than the viral load in the patient's previous sample. In another embodiment, the amount of the substance and / or phenomenon in the first sample is at least 10% lower than the amount of the same substance and / or phenomenon in the second sample. In another embodiment, the amount of substance and / or phenomenon in the first sample is at least 25% lower than the amount of the same substance and / or phenomenon in the second sample. In yet another embodiment, the amount of substance and / or phenomenon in the first sample is at least 50% lower than the amount of the same substance and / or phenomenon in the second sample. In a further embodiment, the amount of substance and / or phenomenon in the first sample is at least 75% lower than the amount of the same substance and / or phenomenon in the second sample. In yet another embodiment, the amount of substance and / or phenomenon in the first sample is at least 90% lower than the amount of the same substance and / or phenomenon in the second sample.

[0130] As used herein, the terms “salt” and “prodrug” include any pharmaceutically acceptable salt, ester, hydrate, or other compound that, upon administration to a recipient, can (directly or indirectly) provide the compound of the invention or its active metabolite or residue. The term “pharmaceutically acceptable salt” refers to, but is not limited to, derivatives of the disclosed compounds, wherein the parent compound is modified by converting an existing acid or base moiety into its salt form (e.g., by reacting a free base group with a suitable organic acid). Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues such as amines; basic or organic salts of acidic residues such as carboxylic acids; etc. Representative acid addition salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, hydrogen sulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, fumarate, gluconate, glyceryl phosphate, hemisulfate, heptasulfate, hexanoate, hydrobromide, hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pyrate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, neopentanoate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, toluenesulfonate, undecanoate, and valerate, etc. Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc., as well as non-toxic ammonium, quaternary ammonium, and amine cations, including, but not limited to, ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, etc. Pharmaceutically acceptable salts of the present invention include conventional non-toxic salts of parent compounds, for example, formed from non-toxic inorganic or organic acids. Pharmaceutically acceptable salts can be synthesized from parent compounds containing basic or acidic moieties using conventional chemical methods. Typically, such salts can be prepared by reacting the free acidic or basic form of these compounds with a stoichiometric amount of a suitable base or acid in water, in an organic solvent, or in a mixture of both; typically, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. In a particular embodiment, the salt is a sodium salt. A list of suitable salts can be found in the following literature: for example, Remington: The Science and Practice of PharmacyLoyd V. Allen, Jr (Ed), The Pharmaceutical Press, London, 22nd ed., September 2012; Stahl and Wermuth (2002) Pharmaceutical Salts: Properties, Selection, and Use, Wiley-VCH; and Berge et al. (1977). Journal of Pharmaceutical Science , 66: 1-19, each of which is incorporated into this text in its entirety by reference.

[0131] As used in this article, the term "systemic inflammatory response syndrome" or "SIRS" refers to a clinical response to nonspecific injury that is characterized by two or more of the following measurable clinical features: body temperature greater than 38°C or less than 36°C, heart rate greater than 90 beats per minute, respiratory rate greater than 20 breaths per minute, and white blood cell count (total white blood cells) greater than 12,000 / mm³. 3 or less than 4,000 / mm 3 An infection is indicated by a neutrophil percentage band greater than 10%. From an immunological perspective, this can be considered a systemic response to infectious (e.g., pathogenic microorganisms) or non-infectious injury (e.g., major surgery) or systemic inflammation. Confirmation of infection can be determined using any suitable procedure known in the art, exemplary examples of which include blood culture, nucleic acid detection (e.g., PCR, mass spectrometry), immunological detection (e.g., ELISA), isolation of bacteria from infected cells, cell lysis, and imaging techniques (such as electron microscopy).

[0132] As used herein, the terms “treatment”, “treating”, etc., refer to the attainment of desired pharmacological and / or physiological effects. These effects may be therapeutic, manifested as partial or complete cure of a disease or condition (e.g., a viral infection) and / or adverse effects caused by said disease or condition. These terms also cover any treatment of a condition or disease in a subject (particularly a human), and include: (a) suppressing the disease or condition, i.e., preventing its development; or (b) alleviating the disease or condition, i.e., causing the remission of the disease or condition.

[0133] Unless otherwise specified, each implementation scheme described herein is applicable to each and every implementation scheme after necessary modifications to the details.

[0134] 2. Compounds

[0135] This invention is based in part on the discovery of specific compounds that bind to viral fusion proteins and inhibit the interaction between the fusion proteins and host cells. Therefore, the inventors contemplate that such compounds could be used to treat and inhibit the development of viral infections in subjects, particularly infections of viruses expressing viral fusion proteins, such as enveloped viral infections (e.g., coronavirus infections, such as SARS-CoV-2 infection), or to treat conditions associated with viral infections, such as acute inflammatory conditions, CRS, cytokine storms, or SARS.

[0136] Therefore, one aspect of the present invention provides compounds of formula I, II, or III:

[0137]

[0138] (I)

[0139]

[0140] (II)

[0141]

[0142] (III)

[0143] Or its pharmaceutically acceptable salt, solvate, or prodrug,

[0144] in:

[0145] L 1 L 3 L 4 L 6 L 7 L 8 L 9 L 10 L 11 L 12 L 13 L 15 L 16 L 17 L 18 and L 19 Independently selected from optionally substituted C1-C5 alkylene groups, optionally substituted C2-C5 alkenyl groups, and optionally substituted C2-C5 ynylene groups;

[0146] L 2 L 5 and L 14 Independently selected from optionally substituted C2-C8 alkylene groups, optionally substituted C2-C8 alkenyl groups, and optionally substituted C2-C8 ynylene groups;

[0147] X 1 X2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 and X 11 Independently selected from CH and N; and

[0148] R 1 Selected from C2-C which are optionally replaced 12 Alkyl groups, optionally substituted C2-C 12 Alkenyl and optionally substituted C2-C 12 Alkyne group.

[0149] Suitable optional substituents include, but are not limited to, methyl, ethyl, and halogen. In certain embodiments, each optionally substituted alkylene, alkenylene, ynylene, alkyl, alkenyl, or ynyl group may be replaced by 1-3 substituents, particularly 1-3 methyl groups. In some embodiments, the alkylene, alkenylene, ynylene, alkyl, alkenyl, and ynyl groups are unsubstituted.

[0150] In a particular implementation, X 1 -X 11 It is CH. To avoid any ambiguity, when X 1 -X 11 When it is CH, the carbon atom is a member of the ring (i.e., the ring is a triazole).

[0151] In some implementations, L 1 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkyne groups; particularly optionally substituted C1-C3 alkylene groups; more particularly C1-C3 alkylene groups; and most particularly methylene groups.

[0152] In some implementations, L 2 Selected from optionally substituted C2-C5 alkylene, optionally substituted C2-C5 alkenyl, and optionally substituted C2-C5 ynynyl; particularly optionally substituted C2-C5 alkylene; more particularly C2-C5 alkylene; most particularly propylene (e.g., n-propylene).

[0153] In some implementations, L 3 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkyne groups; particularly optionally substituted C1-C3 alkylene groups; more particularly C1-C3 alkylene groups; and most particularly methylene groups.

[0154] In a particular implementation, L 4 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkyne groups; particularly optionally substituted C1-C3 alkylene groups; more particularly C1-C3 alkylene groups; and most particularly methylene groups.

[0155] In some implementations, L 5 Selected from optionally substituted C3-C6 alkylene groups, optionally substituted C3-C6 alkenyl groups, and optionally substituted C6-C6 ynynyl groups; particularly optionally substituted C3-C6 alkylene groups; more particularly C3-C6 alkylene groups; and most particularly butylene groups (e.g., n-butylene groups).

[0156] In a particular implementation, L 6 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkyne groups; particularly optionally substituted C1-C3 alkylene groups; more particularly C1-C3 alkylene groups; and most particularly methylene groups.

[0157] In the specific implementation plan, L 7 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkyne groups; particularly optionally substituted C1-C3 alkylene groups; more particularly C1-C3 alkylene groups; and most particularly methylene groups.

[0158] In some implementations, L 8 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkyne groups; particularly optionally substituted C1-C3 alkylene groups; more particularly C1-C3 alkylene groups; and most particularly methylene groups.

[0159] In some implementations, L 9 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkyne groups; particularly optionally substituted C1-C3 alkylene groups; more particularly C1-C3 alkylene groups; and most particularly methylene groups.

[0160] In a particular implementation, L 10 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkyne groups; particularly optionally substituted C1-C3 alkylene groups; more particularly C1-C3 alkylene groups; and most particularly methylene groups.

[0161] In some implementations, L 11 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkyne groups; particularly optionally substituted C1-C3 alkylene groups; more particularly C1-C3 alkylene groups; and most particularly methylene groups.

[0162] In some implementations, L 12 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkyne groups; particularly optionally substituted C1-C3 alkylene groups; more particularly C1-C3 alkylene groups; and most particularly methylene groups.

[0163] In some implementations, L 13 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkyne groups; particularly optionally substituted C1-C3 alkylene groups; more particularly C1-C3 alkylene groups; and most particularly methylene groups.

[0164] In some implementations, L 14 Selected from optionally substituted C2-C4 alkylene groups, optionally substituted C2-C4 alkenyl groups, and optionally substituted C2-C4 ynynyl groups; particularly optionally substituted C2-C4 alkylene groups; more particularly C2-C4 alkylene groups; and most particularly ethylene groups.

[0165] In some implementations, L 15 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkyne groups; particularly optionally substituted C1-C3 alkylene groups; more particularly C1-C3 alkylene groups; and most particularly methylene groups.

[0166] In some implementations, L 16 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkyne groups; particularly optionally substituted C1-C3 alkylene groups; more particularly C1-C3 alkylene groups; and most particularly methylene groups.

[0167] In some implementations, L 17 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkyne groups; particularly optionally substituted C1-C3 alkylene groups; more particularly C1-C3 alkylene groups; and most particularly methylene groups.

[0168] In some implementations, L18 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkyne groups; particularly optionally substituted C1-C3 alkylene groups; more particularly C1-C3 alkylene groups; and most particularly methylene groups.

[0169] In some implementations, L 19 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkyne groups; particularly optionally substituted C1-C3 alkylene groups; more particularly C1-C3 alkylene groups; and most particularly methylene groups.

[0170] In some implementations, R 1 Selected from C6-C which are optionally replaced 10 Alkyl groups, optionally substituted C6-C 10 Alkenyl and optionally substituted C6-C 10 Alkyne group; especially the optionally substituted C6-C 10 Alkyl groups; more particularly C6-C 10 Alkyl. In a particular embodiment, R 1 Selected from optionally substituted C7-C9 alkyl, optionally substituted C7-C9 alkenyl and optionally substituted C7-C9 ynyl; particularly optionally substituted C7-C9 alkyl; more particularly C7-C9 alkyl; most particularly octyl.

[0171] In a specific implementation, the compound is a compound of formula I:

[0172]

[0173] (I)

[0174] Or its pharmaceutically acceptable salt, solvate, or prodrug,

[0175] in:

[0176] L 1 and L 3 Independently selected from optionally substituted C1-C5 alkylene groups, optionally substituted C2-C5 alkenyl groups, and optionally substituted C2-C5 ynylene groups;

[0177] L 2 Selected from optionally substituted C2-C8 alkylene groups, optionally substituted C2-C8 alkenyl groups, and optionally substituted C2-C8 ynylene groups; and

[0178] X 1 and X 2 Selected independently from CH and N.

[0179] L 1 L 2 L 3 X 1 and X 2 The appropriate implementation scheme for each of them is discussed above.

[0180] In a specific implementation:

[0181] L 1 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkynyl groups;

[0182] L 2 Selected from optionally substituted C2-C5 alkylene groups, optionally substituted C2-C5 alkenyl groups, and optionally substituted C2-C5 ynylene groups;

[0183] L 3 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 ynylene groups; and

[0184] X 1 and X 2 Selected independently from CH and N.

[0185] In other implementation schemes:

[0186] L 1 It is a C1-C3 alkylene group that is optionally substituted;

[0187] L 2 It is an optional substituted C2-C5 alkylene group;

[0188] L 3 It is an optionally substituted C1-C3 alkylene group; and

[0189] X 1 and X 2 Selected independently from CH and N.

[0190] In a specific implementation:

[0191] L 1 It is methylene;

[0192] L 2 It is propylidene (e.g., n-propylidene);

[0193] L 3 It is methylene; and

[0194] X 1 and X 2 It is CH.

[0195] In the specific implementation scheme, the compound of formula I is the compound of formula IV:

[0196]

[0197] (IV)

[0198] Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0199] In a specific implementation, the compound is a compound of formula II:

[0200]

[0201] (II)

[0202] Or its pharmaceutically acceptable salt, solvate, or prodrug,

[0203] in:

[0204] L 4 L 6 L 7 and L 8 Independently selected from optionally substituted C1-C5 alkylene groups, optionally substituted C2-C5 alkenyl groups, and optionally substituted C2-C5 ynylene groups;

[0205] L 5 Selected from optionally substituted C2-C8 alkylene groups, optionally substituted C2-C8 alkenyl groups, and optionally substituted C2-C8 ynylene groups;

[0206] X 3 X 4 and X 5 Independently selected from CH and N; and

[0207] R 1 Selected from C2-C which are optionally replaced 12 Alkyl groups, optionally substituted C2-C 12 Alkenyl and optionally substituted C2-C 12 Alkyne group.

[0208] L 4 L 5 L 6 L 7 L 8 X 3 X 4 X 5 and R 1 The appropriate implementation scheme for each of them is discussed above.

[0209] In a specific implementation:

[0210] L 4 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkynyl groups;

[0211] L 5 Selected from optionally substituted C3-C6 alkylene groups, optionally substituted C3-C6 alkenyl groups, and optionally substituted C6-C6 ynylene groups;

[0212] L 6 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkynyl groups;

[0213] L 7 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkynyl groups;

[0214] L 8 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkynyl groups;

[0215] X 3 X 4 and X 5 Independently selected from CH and N; and

[0216] R 1 Selected from C6-C which are optionally replaced 10 Alkyl groups, optionally substituted C6-C 10 Alkenyl and optionally substituted C6-C 10 Alkyne group.

[0217] In a specific implementation:

[0218] L 4 It is a C1-C3 alkylene group that is optionally substituted;

[0219] L 5 It is an optional substituted C3-C6 alkylene group;

[0220] L 6 It is a C1-C3 alkylene group that is optionally substituted;

[0221] L 7 It is a C1-C3 alkylene group that is optionally substituted;

[0222] L 8 It is a C1-C3 alkylene group that is optionally substituted;

[0223] X 3 X4 and X 5 Independently selected from CH and N; and

[0224] R 1 C6-C is arbitrarily replaced. 10 alkyl.

[0225] In a specific implementation:

[0226] L 4 It is methylene;

[0227] L 5 It is butylene (e.g., n-butylene);

[0228] L 6 It is methylene;

[0229] L 7 It is methylene;

[0230] L 8 It is methylene;

[0231] X 3 X 4 and X 5 It is CH; and

[0232] R 1 It is a C7-C9 alkyl group, especially an octyl group.

[0233] In a particular embodiment, the compound of formula II is the compound of formula V:

[0234]

[0235] (V)

[0236] Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0237] In an alternative embodiment, the compound is a compound of formula III:

[0238]

[0239] (III)

[0240] Or its pharmaceutically acceptable salt, solvate, or prodrug,

[0241] in:

[0242] L 9 L 10 L 11 L 12 L 13 L 15 L 16 L17 L 18 and L 19 Independently selected from optionally substituted C1-C5 alkylene groups, optionally substituted C2-C5 alkenyl groups, and optionally substituted C2-C5 ynylene groups;

[0243] L 14 Selected from optionally substituted C2-C8 alkylene groups, optionally substituted C2-C8 alkenyl groups, and optionally substituted C2-C8 ynylene groups; and

[0244] X 6 X 7 X 8 X 9 X 10 and X 11 Selected independently from CH and N.

[0245] L 9 L 10 L 11 L 12 L 13 L 14 L 15 L 16 L 17 L 18 L 19 X 6 X 7 X 8 X 9 X 10 and X 11 The appropriate implementation scheme for each of them is discussed above.

[0246] In a specific implementation:

[0247] L 9 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkynyl groups;

[0248] L 10 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkynyl groups;

[0249] L 11 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkynyl groups;

[0250] L 12Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkynyl groups;

[0251] L 13 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkynyl groups;

[0252] L 14 Selected from optionally substituted C2-C4 alkylene groups, optionally substituted C2-C4 alkenyl groups, and optionally substituted C2-C4 ynylene groups;

[0253] L 15 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkynyl groups;

[0254] L 16 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkynyl groups;

[0255] L 17 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkynyl groups;

[0256] L 18 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 alkynyl groups;

[0257] L 19 Selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 ynylene groups; and

[0258] X 6 X 7 X 8 X 9 X 10 and X 11 Selected independently from CH and N.

[0259] In the specific implementation plan:

[0260] L 9 It is a C1-C3 alkylene group that is optionally substituted;

[0261] L 10 It is a C1-C3 alkylene group that is optionally substituted;

[0262] L 11It is a C1-C3 alkylene group that is optionally substituted;

[0263] L 12 It is a C1-C3 alkylene group that is optionally substituted;

[0264] L 13 It is a C1-C3 alkylene group that is optionally substituted;

[0265] L 14 It is a C2-C4 alkylene group that is optionally substituted;

[0266] L 15 It is a C1-C3 alkylene group that is optionally substituted;

[0267] L 16 It is a C1-C3 alkylene group that is optionally substituted;

[0268] L 17 It is a C1-C3 alkylene group that is optionally substituted;

[0269] L 18 It is a C1-C3 alkylene group that is optionally substituted;

[0270] L 19 It is an optionally substituted C1-C3 alkylene group; and

[0271] X 6 X 7 X 8 X 9 X 10 and X 11 Selected independently from CH and N.

[0272] In some implementation schemes:

[0273] L 9 It is methylene;

[0274] L 10 It is methylene;

[0275] L 11 It is methylene;

[0276] L 12 It is methylene;

[0277] L 13 It is methylene;

[0278] L 14 It is ethylene;

[0279] L 15 It is methylene;

[0280] L 16 It is methylene;

[0281] L 17 It is methylene;

[0282] L 18 It is methylene;

[0283] L 19 It is methylene; and

[0284] X 6 X 7 X 8 X 9 X 10 and X 11 It is CH.

[0285] In some embodiments, the compound of formula III is a compound of formula VI:

[0286]

[0287] (VI)

[0288] Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0289] In certain embodiments, the present invention provides compounds of formula I, II, III, IV, V, or VI, or pharmaceutically acceptable salts thereof. In some embodiments, the salt is a potassium, lithium, or sodium salt, particularly a sodium salt.

[0290] This document provides compounds of Formula I or pharmaceutically acceptable salts, solvates, or prodrugs thereof. In some embodiments, the invention provides compounds of Formula I or pharmaceutically acceptable salts thereof. In some embodiments, the salt is a potassium, lithium, or sodium salt, particularly a sodium salt.

[0291] This document further provides compounds of formula II or pharmaceutically acceptable salts, solvates, or prodrugs thereof. In some embodiments, the invention provides compounds of formula II or pharmaceutically acceptable salts thereof. In some embodiments, the salt is a potassium, lithium, or sodium salt, particularly a sodium salt.

[0292] In another aspect, compounds of formula III or pharmaceutically acceptable salts, solvates, or prodrugs thereof are provided. In some embodiments, the invention provides compounds of formula III or pharmaceutically acceptable salts thereof. In some embodiments, said salt is a potassium, lithium, or sodium salt, particularly a sodium salt.

[0293] As discussed above, the present invention provides compounds of formula IV or pharmaceutically acceptable salts, solvates, or prodrugs thereof. In some embodiments, the present invention provides compounds of formula IV or pharmaceutically acceptable salts thereof. In some embodiments, the salt is a potassium, lithium, or sodium salt, particularly a sodium salt.

[0294] This document further provides compounds of formula V or pharmaceutically acceptable salts, solvates, or prodrugs thereof. In some embodiments, the invention provides compounds of formula V or pharmaceutically acceptable salts thereof. In some embodiments, said salt is a potassium, lithium, or sodium salt, particularly a sodium salt.

[0295] In another aspect, compounds of formula VI or pharmaceutically acceptable salts, solvates, or prodrugs thereof are provided. In some embodiments, the invention provides compounds of formula VI or pharmaceutically acceptable salts thereof. In some embodiments, said salt is a potassium, lithium, or sodium salt, particularly a sodium salt.

[0296] This document also discloses compounds of formula VII in the method of the present invention:

[0297]

[0298] (VII)

[0299] Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0300] In some embodiments, the compound of formula VII is in the form of a pharmaceutically acceptable salt, such as a potassium, lithium, or sodium salt, especially a sodium salt.

[0301] In some embodiments, the compounds of the present invention are G4, G19, G22 or G24; particularly G19, G22 or G24; most particularly G24.

[0302] The compounds of the present invention may also be in the form of pharmaceutically acceptable salts, solvates, or prodrugs. In some embodiments, the compounds may be in the form of hydrates.

[0303] While any suitable salt is considered, in some embodiments the compounds of the present invention are in the form of pharmaceutically acceptable salts. In particular embodiments, the salt is a potassium, lithium, or sodium salt, especially a sodium salt. Those skilled in the art will understand methods for preparing suitable salts of the compounds of the present invention.

[0304] In some embodiments, the prodrug is an ester, such as O - Neopentyl ester (e.g., described in Gordon et al. (2018)) J Med Chem, (The entire contents of 61: 10340-10344 are incorporated herein by reference).

[0305] The compound can be prepared using techniques known in the art, such as the methods disclosed in WO 2018 / 068090 A1 (the entire contents of which are incorporated herein by reference).

[0306] 3. Composition

[0307] While the compounds of the present invention may be administered in undiluted form, it is preferred that such compounds be provided in the form of pharmaceutical compositions. Therefore, in some embodiments, the compounds may be in the form of pharmaceutical compositions comprising the compounds of the present invention along with a pharmaceutically acceptable carrier or diluent.

[0308] The compound can be formulated into pharmaceutical compositions as a neutral or salt form, particularly as a sodium salt.

[0309] Those skilled in the art will understand that the choice of pharmaceutically acceptable carrier or diluent will depend on the route of administration and the nature of the condition to be treated and the subject. A particular carrier or delivery system and route of administration can be readily determined by those skilled in the art. Care should be taken in selecting the carrier or delivery system and route of administration to ensure that the activity of the compound is not depleted during formulation preparation and that the compound reaches the site of action intact. The pharmaceutical compositions of the present invention can be administered via a variety of routes, including, but not limited to, oral, rectal, topical, intranasal, inhalation, intraocular, transmucosal, enteric, intramuscular, subcutaneous, intramedullary, intrathecal, intravenous, intracerebral, intravaginal, intravesical, intravenous, or intraperitoneal administration. In certain embodiments, the pharmaceutical compositions are administered orally, inhaled, intranasally, topically, or intravenously, particularly orally, inhaled, or intranasally.

[0310] Suitable drug forms for injection include sterile solutions or dispersions, as well as sterile powders for preparing sterile solutions. Such forms should remain stable under production and storage conditions and be protected against reduction, oxidation, and microbial contamination.

[0311] Those skilled in the art will be able to readily determine suitable formulations of the compound using conventional methods. Formulation and administration techniques can be found, for example, Remington: The Science and Practice of Pharmacy Loyd V. Allen, Jr (Ed), The Pharmaceutical Press, London, 22nd edition, September 2012.

[0312] The determination of preferred pH ranges and suitable excipients (such as antioxidants) is a routine technique in the art, for example, as described in Katdare and Chaubel (2006) Excipient Development for Pharmaceutical, Biotechnology and Drug Delivery Systems (CRC Press). Buffer systems are routinely used to provide a desired pH range and may include, but are not limited to, carboxylic acid buffers such as acetate, citrate, lactate, tartrate, and succinate; glycine; histidine; phosphate; tris(hydroxymethyl)aminomethane (Tris); arginine; sodium hydroxide; glutamate; and carbonate buffers. Suitable antioxidants may include, but are not limited to, phenolic compounds such as butylated hydroxytoluene (BHT) and butylated hydroxyanisole; vitamin E; ascorbic acid; reducing agents such as methionine or sulfites; metal chelating agents such as ethylenediaminetetraacetic acid (EDTA); cysteine ​​hydrochloride; sodium bisulfite; sodium metabisulfite; sodium sulfite; ascorbyl palmitate; lecithin; propyl gallate; and α-tocopherol.

[0313] For injection, the compound can be formulated as an aqueous solution, appropriately in a physiologically compatible buffer such as Hanks' solution, Ringer's solution, dextran solution, or a saline buffer such as phosphate-buffered saline (PBS). For transmucosal administration, an exfoliating agent suitable for the barrier to be penetrated is used in the formulation. Such exfoliating agents are well known in the art.

[0314] The compositions of the present invention can be formulated for application in liquid form containing an acceptable diluent (such as saline and sterile water), or can be in the form of a lotion, cream, or gel containing an acceptable diluent or carrier to impart a desired texture, consistency, viscosity, and appearance. Acceptable diluents and carriers are well known to those skilled in the art and include, but are not limited to, ethoxylated and nonethoxylated surfactants, fatty alcohols, fatty acids, hydrocarbon oils (such as palm oil, coconut oil, and mineral oil), cocoa butter wax, silicone oil, pH adjusters, cellulose derivatives, emulsifiers such as nonionic organic and inorganic bases, preservatives, wax esters, sterols, triglycerides, phospholipids such as lecithin and cephalin, polyol esters, fatty alcohol esters, hydrophilic lanolin derivatives, and hydrophilic beeswax derivatives.

[0315] Alternatively, the compounds can be readily formulated into dosages suitable for oral administration using pharmaceutically acceptable carriers well known in the art, which is also considered for the practice of this invention. Such carriers enable the compounds of this invention to be formulated into dosage forms such as tablets, pills, capsules, liquids, gels, syrups, pastes, suspensions, etc., for oral ingestion by patients to be treated. These carriers can be selected from sugars, chitosan, starch, cellulose and its derivatives, malt, gelatin, talc, calcium sulfate, vegetable oils, synthetic oils, polyols, alginic acid, phosphate buffer solutions, emulsifiers, isotonic saline, and pyrogen-free water.

[0316] Pharmaceutical formulations for parenteral administration include aqueous solutions of the composition in an aqueous form. Furthermore, suspensions of the compounds can be prepared as suitable oily injectable suspensions. Suitable lipophilic solvents or mediators include fatty oils such as sesame oil or synthetic fatty acid esters such as ethyl oleate or triglycerides. Aqueous injectable suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers or agents that improve the solubility of the compounds to allow for the preparation of highly concentrated solutions.

[0317] Sterile solutions can be prepared by combining the desired amount of the compound in a suitable solvent with other desired excipients as described above, followed by sterilization, such as by filtration. Dispersions are typically prepared by incorporating various sterilized active compounds into a sterile medium containing a basic dispersion medium and the desired excipients as described above. Sterile dry powders can be prepared by vacuum drying or freeze-drying the sterile solution containing the active compound and other desired excipients as described above.

[0318] Pharmaceutical articles for oral use can be obtained by combining the compound with a solid excipient and processing the granular mixture after adding a suitable excipient (if desired) to obtain tablets or sugar-coated pellet cores. Suitable excipients are specifically fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose products such as, for example, corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, hydroxypropyl methylcellulose, sodium carboxymethylcellulose, and / or polyvinylpyrrolidone (PVP). If desired, disintegrants such as cross-linked polyvinylpyrrolidone, agar, or alginate or its salts, such as sodium alginate, can be added. Such compositions can be prepared by any pharmaceutical method, but all methods include the step of combining one or more therapeutic agents as described above with a carrier constituting one or more essential components. Generally, the pharmaceutical compositions of the present invention can be prepared in ways known per se, for example by conventional methods of mixing, dissolving, granulating, making sugar-coated pellets, grinding, emulsifying, encapsulating, sealing, or low-pressure lyophilizing.

[0319] A suitable coating can be provided for the sugar-coated pellet core. For this purpose, a concentrated sugar solution can be used, which may optionally contain gum arabic, talc, polyvinylpyrrolidone, polycarboxylate gel, polyethylene glycol and / or titanium dioxide, lacquer solution, and suitable organic solvents or solvent mixtures. Dyes or pigments can be added to the coating of tablets or sugar-coated pellets to identify or characterize different combinations of particle dosages.

[0320] Orally administered medications include push-in capsules made of gelatin and soft-sealable capsules made of gelatin and plasticizers (such as glycerin or sorbitol). The push-in capsules may contain an active ingredient mixed with: a filler, such as lactose; a binder, such as starch; and / or a lubricant, such as talc or magnesium stearate; and optionally a stabilizer. In soft capsules, the active compound may be dissolved or suspended in a suitable liquid, such as fatty oil, liquid paraffin, or liquid polyethylene glycol. Additionally, a stabilizer may be added.

[0321] The compound can be incorporated into modulated-release products and formulations, such as polymer microsphere formulations and oil- or gel-based formulations.

[0322] The compound may be administered locally rather than systemically, such as by direct injection into tissues, preferably subcutaneous or omental tissues, often in reservoirs or sustained-release formulations. In other embodiments, the compound is administered systemically.

[0323] Furthermore, the compound can be administered in targeted drug delivery systems, such as in particles suitable for targeting cells or tissues and selectively absorbed by them. In some embodiments, the compound is contained in or otherwise bound to a medium selected from liposomes, micelles, dendritic polymers, biodegradable particles, artificial DNA nanostructures, lipid-based nanoparticles, and carbon or gold nanoparticles. In exemplary examples of this type, the medium is selected from poly(lactic acid) (PLA), poly(hydroxyacetic acid) (PGA), poly(lactic-co-hydroxyacetic acid) (PLGA), poly(ethylene glycol) (PEG), PLA-PEG copolymers, and combinations thereof.

[0324] In cases of local application or selective absorption, the effective local concentration of the drug may be independent of the plasma concentration.

[0325] In some embodiments, the composition is suitable for inhalation or intranasal delivery and is in the form of, for example, a solution, aerosol, dry powder, suspension, or emulsion. For example, the composition can be applied to the respiratory tract as a nasal or lung inhalation aerosol or solution for use in nebulizers, or as a fine powder (e.g., particles having a diameter of 1 to 10 μm or less) for inhalation, used alone or in combination with an inert carrier (such as lactose or glucose) or with other pharmaceutically acceptable excipients (such as β-cyclodextrin, starch, sodium carboxymethyl cellulose, etc.).

[0326] Aerosol formulations include those in which the compound is provided together with a suitable propellant in a pressurized package, such as metered-dose inhalers. While the propellant may be a chlorofluorocarbon (CFC) (such as dichlorodifluoromethane, trichlorofluoromethane, or 1,2-dichlorotetrafluoroethane), the propellant is more preferably a non-CFC propellant such as an inert gas (e.g., carbon dioxide, nitrogen, or nitrogen dioxide), a hydrofluoroalkane [e.g., 1,1,1,2-tetrafluoroethane (HFA-134a), 1,1,1,2,3,3,3-heptafluoropropane (HFA-227), 1,1-difluoroethane (HFC-152a), or 1,1,1,2,3,3,3-heptafluoroethane (HFA-227ea)], a hydrochlorofluorocarbon, a hydrofluoroolefin [e.g., 1,3,3,3-tetrafluoroprop-1-ene (HFO-1234ze) or 2,3,3,3-tetrafluoroprop-1-ene (HFO-1234yf)], or a compressed gas (e.g., compressed air). The aerosol can also conveniently contain surfactants such as lecithin. The dosage of the compound can be controlled by setting a metering valve. It has been found that a particle size of approximately 1 to 5 μm is suitable for delivery to the lungs, because particles smaller than 1 μm are usually exhaled and cannot be delivered to the lungs, and particles larger than 10 μm are mostly trapped by oropharyngeal deposits and cannot reach the lungs. Devices driven by HFA-134a deliver even smaller droplets, which are more easily penetrated into the bronchial airways. For drug delivery via the nasal passages, a suitable particle size is, for example, 20–80 μm, because smaller particles (less than 10 μm) are carried into the tracheobronchial region, while larger particles (greater than 100 μm) are rapidly cleared from the nasal passages.

[0327] The compound can also be provided in pharmaceutical formulations that form a gel within the nasal cavity. The compound can also be formulated as a powder composition, which can be presented in unit doses, for example in capsules or tubes containing gelatin, or in blister packs, from which the powder can be administered by means of an inhaler.

[0328] For ease of application and uniform dosage, it is advantageous to formulate the composition in dosage unit form. The determination of the novel dosage unit form of the present invention depends on and is directly dependent on the unique characteristics of the active substance, the specific therapeutic effect to be achieved, and the inherent limitations in the field of active substance formulation for treating diseases in living subjects (where physical health is impaired), as detailed herein.

[0329] While the compounds of the present invention may be the sole active agent administered to a subject, the administration of other active agents in parallel with said compounds is within the scope of the invention. For example, in some embodiments, said compounds may be administered in parallel with one or more antimicrobial agents, anti-inflammatory agents, or agents that inhibit cytokine storms. The compounds may be used therapeutically after other active agents, or may be used therapeutically together with other active agents. The compounds may be administered separately from, simultaneously with, or sequentially with other active agents.

[0330] Therefore, in another aspect of the invention, a composition is provided comprising the compounds of the invention (such as compounds of formula I, II, III, IV, V, VI or VII) and an antimicrobial agent, an anti-inflammatory agent or an agent that inhibits cytokine storm.

[0331] Suitable antimicrobial agents include, but are not limited to, compounds that kill or inhibit the growth of microorganisms (such as viruses, bacteria, yeasts, fungi, protozoa, etc.), and therefore include antibiotics, antifungal agents, antiprotozoal agents, antimalarial agents, antituberculosis agents, and antiviral agents. In a particular embodiment, the antimicrobial agent is an antiviral agent.

[0332] Exemplary antibiotics include quinolones (e.g., amloxacin, sinofloxacin, ciprofloxacin, enoxacin, fleroxacin, flumethin, lomifloxacin, nalidixic acid, norfloxacin, ofloxacin, levofloxacin, lomifloxacin, oxoryl acid, pefloxacin, rosofloxacin, temafloxacin, tofloxacin, sparfloxacin, clinfloxacin, gatifloxacin, moxifloxacin; gemifloxacin; and canofloxacin), tetracyclines, glycylcyclines, and oxazolidinones (e.g., chlortetracycline, demethylchlorocycline, doxycycline, lysylcycline, methacycline, minocycline). Oxytetracycline, tetracycline, tigecycline; linezolid, epirazolid), glycopeptides, aminoglycosides (e.g., amikacin, abekacin, butirocin, dibekacin, fotiamcinol, gentamicin, kanamycin, neomycin (meomycin), netilmicin, ribostamycin, sisomicin, spectinomycin, streptomycin, tobramycin), β-lactams (e.g., imipenem, meropenem, biapenem, cefaclor, cefadroxil, cefamandole, ceftriaxone, cefoxitin, cefazolin, cefixime, cephalosporin). Oxime, Cefotaxime, Cefonisin, Cefoperazone, Cefradine, Cefotaxime, Cefotiam, Cefimazole, Cefpirome, Cefpodoxime, Cefsulfuron-methyl, Ceftazidime, Cefotelan, Cefotetrazol, Cefbuprofen, Cefazolin, Cefuroxime, Cefazolin, Cefalexin, Cefazolin, Cefotaxime, Cefotaxime, Cefepime, Cefrapne, Cefradix, Cefmetazole, Cefoxidin, Cefotetan, Assuamine, Carrumonam, Fluoxetine, Latamoxef, Mecillin, Amoxicillin, Ampicillin, Aloxicillin Carbenicillin, penicillin, carfilcillin, cloxacillin, dicloxacillin, methicillin, mezlocillin, nafcillin, oxacillin, penicillin G, piperacillin, sulfamethoxazole, temoxicillin, ticarcillin, cefotaxime, SC004, KY-020, cefdinir, cefbufen, FK-312, S-1090, CP-0467, BK-218, FK-037, DQ-2556, FK-518, cefazolin, ME1228, KP-736, CP-6232, Ro 09-1227, OPC-20000, LY206763), rifamycin, macrolides (e.g., azithromycin, clarithromycin, erythromycin, rotamycin, roxamisin, roxithromycin, acetosine), ketone lactones (e.g., telithromycin, cerethromycin), coumarin, lincosamides (e.g., clindamycin, lincomycin), and chloramphenicol.Representative antiviral agents include abacavir sulfate, acyclovir sodium, amantadine hydrochloride, ampravir, cidofovir, deraviridine mesylate, norinosine, efavirenz, famciclovir, fomivirex sodium, foscarnet sodium, ganciclovir, indinavir sulfate, lamivudine, lamivudine / zidovudine, nelfinavir mesylate, nevirapine, oseltamivir phosphate, ribavirin, amantadine hydrochloride, ritonavir, saquinavir, saquinavir mesylate, stavudine, valacyclovir hydrochloride, valacyclovir, zalcitabine, zanamivir, and zidovudine. Non-limiting examples of antiviral agents include atovaquinone, chloroquine hydrochloride, chloroquine phosphate, metronidazole, metronidazole hydrochloride, and pentamidine hydroxyethylsulfonate. Suitable anthelmintics may be selected from at least one of mebendazole, pyrantel pamoate, albendazole, ivermectin, and thiabendazole. Exemplary antifungal agents may be selected from amphotericin B, amphotericin B cholesterol sulfate complex, amphotericin B lipid complex, amphotericin B liposome, bifonazole, butconazole, clodextrin, chlorpheniramine, ciclopirox olamine, clotrimazole, epconazole, econazole, fluconazole, flucytosine, flutramazole, micronized griseofulvin, ultramicronized griseofulvin, itraconazole, isoconazole, itraconazole, ketoconazole, miconazole, nitrofurantoin oxime, nystatin, terbinafine hydrochloride, thiaconazole, teconazole, and undecenoic acid. Non-limiting examples of antimalarial agents include chloroquine hydrochloride, chloroquine phosphate, doxycycline, hydroxychloroquine sulfate, mefloquine hydrochloride, primaquine phosphate, pyrimethamine, and combination preparations of pyrimethamine and sulfadoxine. Antituberculosis agents include, but are not limited to, clofazimine, cycloserine, dapsone, ethambutol hydrochloride, isoniazid, pyrazinamide, rifabutin, rifampin, rifapentine, and streptomycin sulfate.

[0333] In some embodiments, the co-activator is an anti-inflammatory agent, representative examples of which include steroidal anti-inflammatory agents, such as, but not limited to, compounds containing a 17-carbon 4-cyclic system, including sterols, various hormones (as proteolytic steroids), and glycosides. Representative examples of steroidal anti-inflammatory drugs include, but are not limited to, corticosteroids such as hydrocortisone, hydroxytriamcione, alpha-methyldexamethasone, dexamethasone phosphate, beclomethasone dipropionate, clobetasol valerate, desonide, deoxymethasone, desoxycorticosterone acetate, dexamethasone, diclomethasone valerate, diflumethasone hydrochloride, fluclosonide, flumethasone neovalerate, fluocinonide, fluocinonide acetate, fluocinonide acetonide, fluocinonide butyl acetate, fluocinonide, fluprednidene acetate, fluprednylidene acetate, fluocinonide, halcinonide, hydrocortisone acetate, hydrocortisone butyrate, methylprednisolone, triamcinolone, cortisone, todoxaconide, flucetonide, difluorosone diacetate, fluradrenolone, diflorosone diacetate, fluradrenolone Acetone, methylphenidate, amcinafel, acetonide, betamethasone and its ester balances, chlorprednisolone, chlorprednisolone acetate, chlorcotropene, clescinolone, dichlorophenidate, difluoroprednisolone, fluclofenoxuron, fluopyrone, fluprednisolone, hydrocortisone valerate, hydrocortisone cyclopentylpropionate, hydrocortisone ester, methylprednisolone, peramisone, prednisolone, prednisolone, beclomethasone dipropionate, triamcinolone, and mixtures thereof.

[0334] Alternatively, the anti-inflammatory agent may be a nonsteroidal anti-inflammatory agent, non-limiting examples of which include agents that are similar in action to aspirin, including, but not limited to, ibuprofen (ADVIL), naproxen sodium (ALEVE), and acetaminophen (TYLENOL). Other examples of nonsteroidal anti-inflammatory agents include, but are not limited to, oxacins such as piroxicam, itoxacin, tenoxacin, sudoxicam, and CP-14,304; salicylsalicylic acid, benorilate, trisalicylic acid, safapryn, solprin, diflunisal, and fendusal; and acetic acid derivatives such as diclofenac, fenclofenac, indomethacin, sulindac, tometidine, itococcal acid, furofenac, thiophene, zidomexin, acimetacin, fentiac, zoleic acid, and cyclohexane. Clodinic acid, oxifenesin, biphenylacetic acid, and ketorolac; fenamic acid esters, such as mefenamic acid, meclofenamic acid, flufenamic acid, niflufenic acid, and tofenamic acid; propionic acid derivatives, such as benzoxaprofen, flurbiprofen, ketoprofen, fenprofen, fenbufen, indobuprofen, pirprofen, carprofen, oxaprazin, pranoprofen, imidoprofen, thioprofen, sulprofen, aminprofen, and tiprofenic acid; pyrazoles, such as phenylbutazone, hydroxybutazine, fepranoprofen, azaprofen, and trimethoprim. Mixtures of these nonsteroidal anti-inflammatory agents, as well as dermatologically acceptable salts and esters of these agents, may also be used. For example, etofenadyl (a flufenamic acid derivative) is particularly suitable for topical application.

[0335] In other embodiments, the anti-inflammatory agent includes, but is not limited to, transforming growth factor-β3 (TGF-β3), anti-tumor necrosis factor-α (TNF-α) agents, inhibitors or antagonists of IL-6 or IL-6 receptors, IL-1 receptors, IL-1β, TNF, GM-CSF, IFN-γ, JAK-STAT signaling, CCR2, CCR5, complement component C5, IRAK4 and M-CSF receptors, or combinations thereof.

[0336] The compounds of this invention can also be administered in parallel with agents that inhibit cytokine storms. Suitable agents include, for example, compounds that target essential immune pathways, such as the chemokine network and the cholinergic anti-inflammatory pathway. For example, JAK inhibitors (such as JAK 1 and JAK 2 inhibitors) can inhibit cytokine storms and, in some cases, also have antiviral activity. Representative JAK inhibitors include those disclosed in U.S. Patent No. 10,022,378, such as Jakafi, tofacitinib, and baricitinib, as well as LY3009104 / INCB28050, paricitinib / SB1518, VX-509, GLPG0634, INC424, R-348, CYT387, TG 10138, AEG 3482, and their pharmaceutically acceptable salts and prodrugs. Further examples include CEP-701 (letotrinib), AZD1480, INC424, R-348, CYT387, TG 10138, and AEG. 3482, 7-iodo-N-(4-morpholinophenyl)thieno[3,2-d]pyrimidin-2-amine, 7-(4-aminophenyl)-N-(4-morpholinophenyl)thieno[3,2-d]pyrimidin-2-amine, N-(4-(2-(4-morpholinophenylamino)thieno[3,2-d]pyrimidin-7-yl)phenyl)acrylamide, 7-(3-aminophenyl)-N-(4-morpholinophenyl)thieno[3,2-d]pyrimidin-2-amine, N-(3-(2-(4-morpholinophenylamino)thieno[3,2-d]pyrimidin-7-yl)phenyl)acrylamide, N-(4-morpholinophenyl)thieno[3,2-d]pyrimidin-2-amine, 2-( 4-morpholinophenylamino)thieno[3,2-d]pyrimidin-7-carboxylic acid methyl ester, N-(4-morpholinophenyl)-5H-pyrrolo[3,2-d]pyrimidin-2-amine, 7-(4-amino-3-methoxyphenyl)-N-(4-morpholinophenyl)thieno[3,2-d]pyrimidin-2-amine, 4-(2-(4-morpholinophenylamino)thieno[3,2-d]pyrimidin-7-yl)benzenesulfonamide, N,N-dimethyl-3-(2-(4-morpholinophenylamino)thieno[3,2-d]pyrimidin-7-yl)benzenesulfonamide, 1-ethyl-3-(2-methoxy-4-(2-(4-morpholinophenylamino)thieno[3,2-d]pyrimidin- -7-yl)phenyl)urea, N-(4-(2-(4-morpholinophenylamino)thieno[3,2-d]pyrimidin-7-yl)phenyl)methanesulfonamide, 2-methoxy-4-(2-(4-morpholinophenylamino)thieno[3,2-d]pyrimidin-7-yl)phenol, 2-cyano-N-(3-(2-(4-morpholinophenylamino)thieno[3,2-d]pyrimidin-7-yl)phenyl)acetamide, N-(cyanomethyl)-2-(4-morpholinophenylamino)thieno[3,2-d]pyrimidin-7-yl)phenyl)acetamide, N-(cyanomethyl)-2-(4-morpholinophenylamino)thieno[3,2-d]pyrimidin-7-carboxamide, N-(3-(2-(4-morpholinophenylamino)thieno[3,2-d]pyrimidin-7-yl)phenyl)methanesulfonamide, 1-ethyl-3-(4-(2-(4-morpholinophenylamino)thieno[3,2-d]pyrimidin-7-yl)-2-(trifluoromethoxy)phenyl)urea, N-(3-nitrophenyl)-7-phenylthieno[3,2-d]pyrimidin-2-amine, 7-iodo-N-(3-nitrophenyl)thieno[3,2-d]pyrimidin-2-amine, N1-(7-(2-ethylphenyl)thieno[3,2-d]pyrimidin-2-yl)phenyl-1,3-diamine, N-tert-butyl-3-(2-(4-morpholinophenylamino)thieno[3,2-d] Pyrimidin-7-yl)benzenesulfonamide, N1-(7-iodothieno[3,2-d]pyrimidin-2-yl)phenyl-1,3-diamine, 7-(4-amino-3-(trifluoromethoxy)phenyl)-N-(4-morpholinophenyl)thieno[3,2-d]pyrimidin-2-amine, 7-(2-ethylphenyl)-N-(4-morpholinophenyl)thieno[3,2-d]pyrimidin-2-amine, N-(3-(2-(4-morpholinophenylamino)thieno[3,2-d]pyrimidin-7-yl)phenyl)acetamide, N-(cyanomethyl)-N-(3-(2-(4-morpholinophenylamino)thieno[3,2-d]pyrimidin-7-yl)phenyl)methanesulfonamide, N-(cyanomethyl)-N-(4-(2-(4-morpholinophenylamino)thieno[3,2-d]pyrimidin-7-yl)phenyl)methanesulfonamide, N-(cyanomethyl)-N-(4-(2-(4-) 4-(5-methyl-2-(4-morpholinophenylamino)-5H-pyrrolo[3,2-d]pyrimidin-7-yl)phenyl)methanesulfonamide, N-(3-(5-methyl-2-(4-morpholinophenylamino)-5H-pyrrolo[3,2-d]pyrimidin-7-yl)phenyl)methanesulfonamide, 4-(5-methyl-2-(4-morpholinophenylamino)-5H-pyrrolo[3,2-d]pyrimidin-7-yl)benzenesulfonamide, N-(4-(5-methyl-2-(4-morpholinophenylamino)-5H-pyrrolo[3,2-d]pyrimidin-7-yl)phenyl)methanesulfonamide, 7-iodo-N-(4-morpholinophenyl)-5H-pyrrolo[3,2-d]pyrimidin-2-amine, 7-(2-isopropylphenyl)-N-(4-morpholinophenyl)thiophene [3,2-d]pyrimidin-2-amine, 7-bromo-N-(4-morpholinophenyl)thieno[3,2-d]pyrimidin-2-amine, N7-(2-isopropylphenyl)-N2-(4-morpholinophenyl)thieno[3,2-d]pyrimidin-2,7-diamine, N7-(4-isopropylphenyl)-N2-(4-morpholinophenyl)thieno[3,2-d]pyrimidin-2,7-diamine, 7-(5-amino-2-methylphenyl)-N-(4-morpholinophenyl)thieno[3,2-d]pyrimidin-2-amine, N-(cyanomethyl)-4-(2-(4-morpholinophenylamino)thieno[3,2-d]pyrimidin-7-yl)benzamide, 7-iodo-N-(3-morpholinophenyl)thieno[3,2-d]pyrimidin-2-amine2-d]pyrimidin-2-amine, 7-(4-amino-3-nitrophenyl)-N-(4-morpholinophenyl)thieno[3,2-d]pyrimidin-2-amine, 7-(2-methoxypyridin-3-yl)-N-(4-morpholinophenyl)thieno[3,2-d]pyrimidin-2-amine, (3-(7-iodothieno[3,2-d]pyrimidin-2-ylamino)phenyl)methanol, N-tert-butyl-3-(2-(3-morpholinophenylamino)thieno[3,2-d]pyrimidin-7-yl)benzenesulfonamide, N-tert-butyl-3-(2-(3-(hydroxymethyl)phenylamino)thieno[3,2-d]pyrimidin-7-yl)benzenesulfonamide, N-(4-morpholinophenyl)-7-(4- 7-(4-amino-3-nitrophenyl)-N-(3,4-dimethoxyphenyl)thieno[3,2-d]pyrimidin-2-amine, N-tert-butyl-3-(2-(3,4,5-trimethoxyphenylamino)thieno[3,2-d]pyrimidin-7-yl)benzenesulfonamide, 7-(4-amino-3-nitrophenyl)-N-(3,4-dimethoxyphenyl)thieno[3,2-d]pyrimidin-2-amine, N-(3,4-dimethoxyphenyl)-7-(2-methoxypyridin-3-yl)thieno[3,2-d]pyrimidin-2-amine, N-tert-butyl-3-(2-(3,4-dimethoxyphenylamino)thieno[3,2-d]pyrimidin-7-yl)benzenesulfonamide, 7-(2-aminopyrimidin-5-yl)-N-(3,4) N-(3,4-dimethoxyphenyl)-7-(2,6-dimethoxypyridin-3-yl)thieno[3,2-d]pyrimidin-2-amine, N-(3,4-dimethoxyphenyl)-7-(2,4-dimethoxypyridin-5-yl)thieno[3,2-d]pyrimidin-2-amine, 7-iodo-N-(4-(morpholinomethyl)phenyl)thieno[3,2-d]pyrimidin-2-amine, N-tert-butyl-3-(2-(4-(morpholinomethyl)phenylamino)thieno[3,2-d]pyrimidin-7-yl)benzenesulfonamide, 2-cyano-N-(4-methyl-3-(2-(4-morpholinophenylamino)) Thiophene[3,2-d]pyrimidin-7-yl)phenyl)acetamide, ethyl 3-(2-(4-morpholinophenylamino)thiophene[3,2-d]pyrimidin-7-yl)benzoate, 7-bromo-N-(4-(2-(pyrrolidine-1-yl)ethoxy)phenyl)thiophene[3,2-d]pyrimidin-2-amine, N-(3-(2-(4-(2-(pyrrolidine-1-yl)ethoxy)phenylamino)thiophene[3,2-d]pyrimidin-7-yl)phenyl)acetamide, N-(cyanomethyl)-3-(2-(4-morpholinophenylamino)thiophene[3,2-d]pyrimidin-7-yl)benzamide, N-tert-butyl-3-(2-(4-morpholinophenylamino)thiophene[3,2-d]pyrimidin-7-yl)benzamide,2-d]pyrimidin-7-yl)benzamide, N-tert-butyl-3-(2-(4-(1-ethylpiperidin-4-yloxy)phenylamino)thieno-[3,2-d]pyrimidin-7-yl)benzenesulfonamide, tert-butyl-4-(2-(4-(morpholinomethyl)phenylamino)thieno-[3,2-d]pyrimidin-7-yl)-1H-pyrazol-1-carboxylate, 7-bromo-N-(4-((4-ethylpiperazin-1-yl)methyl)phenyl)thieno-[3,2-d]pyrimidin-2-amine, N-tert-butyl-3-(2-(4-((4-ethylpiperazin-1-yl)methyl)phenylamino)- thieno[3,2-d]pyrimidin-7-yl)benzenesulfonamide, N-(4-((4-ethylpiperazin-1-yl)methyl)phenyl)-7-(1H-pyrazol-4-yl)thieno[3,2-d]pyrimidin-2-amine, N-(cyanomethyl)-3-(2-(4-(morpholinomethyl)phenylamino)thieno[3,2-d]pyrimidin-7-yl)benzamide, N-tert-butyl-3-(2-(4-(2-(pyrrolidin-1-yl)ethoxy)phenylamino)thieno[3,2-d]pyrimidin-7-yl)benzenesulfonamide, tert-butyl-3-(2-(4-(2-(pyrrolidin-1-yl)ethoxy)phenylamino)thieno[3,2-d]pyrimidin-7-yl)benzylcarbamate (tert-butyl) pyrrolidin-1-yl)ethoxy)phenylamino)thieno[3,2-d]pyrimidin-7-yl)benzylcarbamate, 3-(2-(4-(2-(pyrrolidin-1-yl)ethoxy)phenylamino)thieno[3,2-d]pyrimidin-7-yl)benzylsulfonamide, 7-(3-chloro-4-fluorophenyl)-N-(4-(2-(pyrrolidin-1-yl)ethoxy)phenyl)thieno[3,2-d]pyrimidin-2-amine, 4-(2-(4-(1-ethylpiperidin-4-yloxy)phenylamino)thieno[3,2-d]pyrimidin-7-yl)-1H-pyrazole-1-carboxylic acid tert-butyl ester, 7-(benzo[d][1,3]dioxacyclopenten-5-yl)-N-( 4-(morpholinomethyl)phenyl)thieno[3,2-d]pyrimidin-2-amine, 5-(2-(4-(morpholinomethyl)phenylamino)thieno[3,2-d]pyrimidin-7-yl)-1H-indole-1-carboxylic acid tert-butyl ester, 7-(2-aminopyrimidin-5-yl)-N-(4-(morpholinomethyl)phenyl)thieno[3,2-d]pyrimidin-2-amine, 4-(2-(-4-(morpholinomethyl)phenylamino)thieno[3,2-d]pyrimidin-7-yl)-5,6-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester, morpholinomethyl)phenylamino)thieno[3,2-d]pyrimidin-7-yl)benzylcarboxylic acid tert-butylmorpholinomethyl)phenylamino)thieno[3,2-d]pyrimidin-7-yl)benzylcarbamate), N-(3-(2-(4-(morpholinomethyl)phenylamino)thieno[3,2-d]pyrimidin-7-yl)phenyl)acetamide, N-(4-(2-(4-(morpholinomethyl)phenylamino)thieno[3,2-d]pyrimidin-7-yl)phenyl)acetamide, N-(3 ... 7-(4-(4-methylpiperazin-1-yl)phenyl)-N-(4-(morpholinomethyl)phenyl)thieno-[3,2-d]pyrimidin-2-amine, N-(2-methoxy-4-(2-(4-(morpholinomethyl)phenylamino)thieno-[3,2-d]pyrimidin-7-yl)phenyl)acetamide, 7-bromo-N-( 3,4,5-Trimethoxyphenyl)thieno[3,2-d]pyrimidin-2-amine, (3-(2-(3,4,5-trimethoxyphenylamino)thieno[3,2-d]pyrimidin-7-yl)phenyl)methanol, (4-(2-(3,4,5-trimethoxyphenylamino)thieno[3,2-d]pyrimidin-7-yl)phenyl)methanol, (3-(2-(4-morpholinophenylamino)thieno[3,2-d]pyrimidin-7-yl)phenyl)methanol (4-(2-(4-morpholinophenylamino)thieno[3,2-d]pyrimidin-7-yl)phenyl)methanol, N-(pyrrolidone-1-yl)ethoxy)phenylamino)thieno[3,2-d]pyrimidin-7-yl)benzyl)methanesulfonamide, morpholinomethyl)phenylamino)thieno[3,2-d]pyrimidin-7-yl)benzylcarbamate tert-butyl morpholinomethyl)phenylamino)thieno[3,2-d]pyrimidin-7-yl)benzylcarbamate, N-(4-(morpholinomethyl)phenyl)-7-(3-(piperazin-1-yl)phenyl)thieno[3,2-d]pyrimidin-2-amine, 7-(6-(2-morpholinoethylamino)pyridin-3-yl)-N-(3,4,5-trimethoxyphenyl)thieno[3,2-d]pyrimidin-2-amine, 7-(2-ethylphenyl)-N-(4-(2-(pyrrolidine- 1-yl)ethoxy)phenyl)thieno[3,2-d]pyrimidin-2-amine, 7-(4-(aminomethyl)phenyl)-N-(4-(morpholinomethyl)phenyl)thieno[3,2-d]pyrimidin-2-amine, N-(4-(1-ethylpiperidin-4-yloxy)phenyl)-7-(1H-pyrazol-4-yl)thieno[3,2-d]pyrimidin-2-amine, N-(2,4-dimethoxyphenyl)-7-phenylthieno[3,2-d]pyrimidin-2-amine, 7-bromo-N-(3,4-dimethoxyphenyl)thieno[3,2-d]pyrimidin-2-amine, N-(3,4-Dimethoxyphenyl)-7-phenylthiopheno[3,2-d]pyrimidine-2-amine and their pharmaceutically acceptable salts and prodrugs.

[0337] Alternatively, or additionally, HMGB1 antibodies and / or COX-2 inhibitors, which downregulate cytokine storms, can be used. Examples of such compounds include Actemra (Roche). Celebrex (celecoxib), a COX-2 inhibitor, can also be used. IL-8 (CXCL8) inhibitors can also be used.

[0338] As described above, the compounds can be formulated into dosage units with suitable pharmaceutically acceptable carriers for convenient and effective administration at an effective amount. Suitable unit doses and maximum daily doses of the compounds of the present invention can be determined based on conventionally used unit doses and maximum daily doses. In some embodiments, the dosage form may contain the compound in amounts ranging from about 0.25 μg to about 2000 mg. The compound may be present in an amount of about 0.25 μg to about 2000 mg / mL of the carrier. In embodiments in which the pharmaceutical composition comprises one or more additional active ingredients, the dosage is determined with reference to the commonly used dosage and administration method of said ingredient.

[0339] 4. How to use

[0340] The compounds of the present invention have been found to interact with viral fusion proteins (e.g., spike proteins) and inhibit the interaction between the fusion proteins and host cells (e.g., the host cell's ACE2). Therefore, the inventors contemplate the use of such compounds for the treatment and inhibition of the development of viral infections in subjects, particularly infections of viruses expressing viral fusion proteins, such as enveloped viral infections (e.g., coronavirus infections, such as SARS-CoV-2 infection), or for the treatment of conditions associated with viral infections (e.g., coronavirus infections), such as acute inflammatory conditions, CRS, or cytokine storms, or SARS. Therefore, compounds of formulas I, II, III, IV, V, or VI, or pharmaceutically acceptable salts, solvates, or prodrugs thereof, are contemplated for use in therapeutics.

[0341] In another aspect, the present invention provides a method for treating or inhibiting the development of a viral infection in a subject, the method comprising, consisting of, or substantially consisting of administering to the subject a compound of formula I, II, III, IV, V, VI, or VII or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

[0342] Further provided are the use of compounds of formula I, II, III, IV, V, VI, or VII, or pharmaceutically acceptable salts, solvates, or prodrugs thereof, for treating or inhibiting the development of a viral infection in a subject; the use of compounds of formula I, II, III, IV, V, VI, or VII, or pharmaceutically acceptable salts, solvates, or prodrugs thereof, for treating or inhibiting the development of a viral infection in a subject; and the use of compounds of formula I, II, III, IV, V, VI, or VII, or pharmaceutically acceptable salts, solvates, or prodrugs thereof, in the preparation of a medicament for treating or inhibiting the development of a viral infection in a subject.

[0343] Suitable embodiments of compounds of formulas I, II, III, IV, V, VI, and VII have been discussed above.

[0344] In some embodiments, the compound is a compound of formula I, II, or III, or a pharmaceutically acceptable salt, solvate, or prodrug thereof; particularly a compound of formula III, or a pharmaceutically acceptable salt, solvate, or prodrug thereof. In some embodiments, the compound is a compound of formula I, II, or III, or a pharmaceutically acceptable salt thereof; particularly a compound of formula III, or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is a compound of formula I, II, or III; particularly a compound of formula III.

[0345] In some embodiments, the compound is a compound of formula IV, V, or VI, or a pharmaceutically acceptable salt, solvate, or prodrug thereof; particularly a compound of formula VI, or a pharmaceutically acceptable salt, solvate, or prodrug thereof. In some embodiments, the compound is a compound of formula IV, V, or VI, or a pharmaceutically acceptable salt thereof; particularly a compound of formula VI, or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is a compound of formula IV, V, or VI; particularly a compound of formula VI.

[0346] While considering the practicality of the compound for treating or inhibiting the development of any viral infection, in certain embodiments, the virus is a virus expressing a viral fusion protein. In some embodiments, the virus is an enveloped virus, particularly an enveloped RNA virus, such as flaviviruses (e.g., dengue virus, West Nile virus, yellow fever virus, Japanese encephalitis virus, St. Louis encephalitis virus, tick-borne encephalitis virus, bovine viral diarrhea virus, and classical swine fever virus), encapsulated viruses (e.g., Sindbis virus, equine encephalitis virus, Chekungunya virus, rubella virus, Ross River virus, bovine diarrhea virus, swine cholera virus, and Simemlik Forest virus), coronaviruses (e.g., beta-coronaviruses such as A-line OC43 and HKU1, porcine gastroenteritis virus, severe acute respiratory syndrome virus (SARS-CoV), and severe acute respiratory syndrome virus 2). (SARS-CoV-2 and Middle East Respiratory Syndrome (MERS)), orthomyxoviruses (such as human influenza viruses, including human influenza A, B, and C types, avian influenza viruses, and equine influenza viruses), paramyxoviruses (such as measles virus, canine distemper virus, mumps virus, parainfluenza virus, respiratory syncytial virus, Newcastle disease virus, rinderpest virus, Nipah virus, and Hendra virus), rhabdoviruses (such as rabies virus, Australian bat rabies virus, and vesicular stomatitis virus), and bunyaviruses (such as hantavirus, LaCrosse virus, and Rift Valley fever). Viruses (such as Marburg virus and Ebola virus); or enveloped DNA viruses, such as herpesviruses (such as herpes simplex virus 1 and 2, varicella-zoster virus, Epstein-Barr virus and human cytomegalovirus), poxviruses (such as smallpox virus, vaccinia virus, cowpox virus, monkeypox virus, rabbitpox virus, contagious oral thrush virus in sheep, pseudococcosis, bovine papular stomatitis virus, Turner poxvirus, Yaba monkey tumor virus and molluscum contagiosum virus), hepatotropic DNA viruses (such as hepatitis B virus) or African swine fever virus family (such as African swine fever virus).

[0347] In certain embodiments, the virus is a coronavirus, specifically said coronavirus capable of causing severe acute respiratory syndrome (SARS). In some embodiments, the coronavirus is a beta-coronavirus, such as one selected from lineage A, lineage B, lineage C, and lineage D. In some embodiments, the beta-coronavirus is lineage B beta-coronavirus, such as SARS-CoV or SARS-CoV-2; particularly SARS-CoV-2. In alternative embodiments, the beta-coronavirus is lineage C beta-coronavirus, such as MERS-CoV.

[0348] In a particular embodiment, the infection is caused by a virus that interacts with cells expressing the ACE2 peptide, for example, when the virus contains a spike protein. In such an embodiment, the virus is an ACE2-binding coronavirus. Suitable coronaviruses include lineage B beta-coronaviruses, such as SARS-CoV or SARS-CoV-2.

[0349] Cells expressing the ACE2 peptide can be any cells expressing ACE2, such as lung cells (e.g., alveolar cells), intestinal cells, endothelial cells, epithelial cells (e.g., nasal or nasopharyngeal epithelial cells), kidney cells (e.g., brush border cells of proximal renal tubular epithelial cells), or arterial smooth muscle cells. In a particular embodiment, the cells are respiratory cells.

[0350] In another aspect, a method for inhibiting the interaction between a virus and cells expressing an ACE2 peptide is provided, the method comprising, consisting of, or substantially consisting of: contacting the virus with a compound of formula I, II, III, IV, V, VI, or VII, or a pharmaceutically acceptable salt, solvate, or prodrug thereof. Also provided are the use of a compound of formula I, II, III, IV, V, VI, or VII, or a pharmaceutically acceptable salt, solvate, or prodrug thereof for inhibiting the interaction between a virus and cells expressing an ACE2 peptide; the use of a compound of formula I, II, III, IV, V, VI, or VII, or a pharmaceutically acceptable salt, solvate, or prodrug thereof for inhibiting the interaction between a virus and cells expressing an ACE2 peptide; and the use of a compound of formula I, II, III, IV, V, VI, or VII, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, in the preparation of a medicament for inhibiting the interaction between a virus and cells expressing an ACE2 peptide.

[0351] In some implementations, the interaction is one or both of virus binding to a cell and virus entry into the cell; particularly virus entry into the cell.

[0352] Suitable embodiments of compounds of formulas I, II, III, IV, V, VI, and VII have been discussed above.

[0353] In some embodiments, the compound is a compound of formula I, II, or III, or a pharmaceutically acceptable salt, solvate, or prodrug thereof; particularly a compound of formula III, or a pharmaceutically acceptable salt, solvate, or prodrug thereof. In some embodiments, the compound is a compound of formula I, II, or III, or a pharmaceutically acceptable salt thereof; particularly a compound of formula III, or a pharmaceutically acceptable salt thereof. In specific embodiments, the compound is a compound of formula I, II, or III; particularly a compound of formula III.

[0354] In some embodiments, the compound is a compound of formula IV, V, or VI, or a pharmaceutically acceptable salt, solvate, or prodrug thereof; particularly a compound of formula VI, or a pharmaceutically acceptable salt, solvate, or prodrug thereof. In some embodiments, the compound is a compound of formula IV, V, or VI, or a pharmaceutically acceptable salt thereof; particularly a compound of formula VI, or a pharmaceutically acceptable salt thereof. In specific embodiments, the compound is a compound of formula IV, V, or VI; particularly a compound of formula VI.

[0355] Cells expressing the ACE2 peptide can be any cells expressing ACE2, such as lung cells (e.g., alveolar cells), intestinal cells, endothelial cells, epithelial cells (e.g., nasal or nasopharyngeal epithelial cells), kidney cells (e.g., brush border cells of proximal renal tubular epithelial cells), or arterial smooth muscle cells. In a particular embodiment, the cells are respiratory cells.

[0356] The above discussed suitable implementation schemes for viruses. In a particular implementation scheme, the virus is a coronavirus, such as SARS-CoV or SARS-CoV-2; especially SARS-CoV-2.

[0357] In another aspect, a method is provided for treating an acute inflammatory condition in a subject, wherein the condition is associated with coronavirus infection, the method comprising, consisting of, or substantially consisting of: administering to the subject a compound of formula I, II, III, IV, V, VI, or VII, or a pharmaceutically acceptable salt, solvate, or prodrug thereof. Also provided is the use of a compound of formula I, II, III, IV, V, VI, or VII, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, for treating an acute inflammatory condition in a subject, wherein the condition is associated with coronavirus infection; and the use of a compound of formula I, II, III, IV, V, VI, or VII, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, for treating an acute inflammatory condition in a subject, wherein the condition is associated with coronavirus infection; and the use of a compound of formula I, II, III, IV, V, VI, or VII, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, in the preparation of a medicament for treating an acute inflammatory condition in a subject, wherein the condition is associated with coronavirus infection.

[0358] Suitable embodiments of compounds of formulas I, II, III, IV, V, VI, and VII have been discussed above.

[0359] In some embodiments, the compound is a compound of formula I, II, or III, or a pharmaceutically acceptable salt, solvate, or prodrug thereof; particularly a compound of formula III, or a pharmaceutically acceptable salt, solvate, or prodrug thereof. In some embodiments, the compound is a compound of formula I, II, or III, or a pharmaceutically acceptable salt thereof; particularly a compound of formula III, or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is a compound of formula I, II, or III; particularly a compound of formula III.

[0360] In some embodiments, the compound is a compound of formula IV, V, or VI, or a pharmaceutically acceptable salt, solvate, or prodrug thereof; particularly a compound of formula VI, or a pharmaceutically acceptable salt, solvate, or prodrug thereof. In some embodiments, the compound is a compound of formula IV, V, or VI, or a pharmaceutically acceptable salt thereof; particularly a compound of formula VI, or a pharmaceutically acceptable salt thereof. In specific embodiments, the compound is a compound of formula IV, V, or VI; particularly a compound of formula VI.

[0361] In certain embodiments, the coronavirus is a beta-coronavirus, such as lineage A beta-coronavirus, lineage B beta-coronavirus, lineage C beta-coronavirus, or lineage D beta-coronavirus. In some embodiments, the beta-coronavirus is lineage B beta-coronavirus, such as SARS-CoV or SARS-CoV-2; particularly SARS-CoV-2. In alternative embodiments, the beta-coronavirus is lineage C beta-coronavirus, such as MERS-CoV. In specific embodiments, the coronavirus is an ACE2-binding coronavirus, such as lineage B beta-coronavirus, including SARS-CoV or SARS-CoV-2.

[0362] In some embodiments, the acute inflammatory condition is associated with the presence of cytokine release syndrome (CRS) or a cytokine storm. Exemplary examples of this type include the CRS or cytokine storm comprising an elevation of at least 50% compared to the baseline state of one or more cytokines or chemokines selected from IFN-γ, IFN-β, TNF-α, IL-1β, IL-6, IL-17A, CCL3, CXCL2, and CXCL8.

[0363] The acute inflammatory condition may be associated with the presence of CRS, for example, in which the subject has one or more symptoms selected from the following: fever, fatigue, loss of appetite, muscle and joint pain, nausea, vomiting, diarrhea, rash, rapid breathing, rapid heartbeat, low blood pressure, seizures, headache, confusion, delirium, hallucinations, tremor, and coordination disorders. The acute inflammatory condition may also be associated with the presence of a cytokine storm, for example, in which the subject may have one or more symptoms selected from the following: high fever, swelling and redness, extreme fatigue, nausea, bleeding, coagulation, visceral organ damage, and shock, or any combination thereof.

[0364] In some implementations, the acute inflammatory condition is associated with multisystem inflammatory syndrome (MIS-C) in children, for example, wherein the subject has one or more symptoms selected from the following: fever, vomiting, diarrhea, stomach pain, rash, red eyes, red or swollen lips and tongue, unusual fatigue, red or swollen hands or feet, severe stomach pain, cardiac symptoms (including chest pain, palpitations, and shortness of breath), cyanosis of lips or face, confusion, inability to be awakened or remain conscious, abdominal pain with vomiting and diarrhea, rash and swelling of the extremities, syncope, and new-onset hypotension.

[0365] The acute inflammatory symptoms can also be associated with systemic inflammatory response syndrome (SIRS). The subjects may have one or more symptoms associated with a specific phase of SIRS, with representative symptoms as follows:

[0366] Phase 1 is a localized response at the site of injury, designed to control the damage and limit its spread. Immune effector cells at this site release cytokines, which in turn stimulate the reticuloendothelial system, promoting wound repair through local inflammation. Nitric oxide and prostacyclin-induced local vasodilation (redness) and disruption of endothelial tight junctions allow leukocytes to marginalize and migrate into the tissue space. Leakage of cells and protein-rich fluid into the extravascular space causes swelling (tumor) and increased heat (fever). Inflammatory mediators affect local somatic sensory nerves, resulting in pain and loss of function. This loss of function also allows the body part to repair itself, rather than continue to be used.

[0367] Phase 2 is the early compensatory anti-inflammatory response syndrome (CARS), designed to maintain immune homeostasis. To maintain homeostasis, growth factor stimulation and recruitment of macrophages and platelets occur as pro-inflammatory mediator levels decrease.

[0368] Stage 3 is a phase that tilts towards pro-inflammatory SIRS, leading to progressive endothelial dysfunction, coagulopathy, and activation of coagulation pathways. It results in microthrombus formation in terminal organs and a gradual increase in capillary permeability, ultimately leading to the loss of circulatory integrity.

[0369] Stage 4 is characterized by the replacement of SIRS by CARS, leading to a state of relative immunosuppression. Consequently, individuals become susceptible to secondary or nosocomial infections, thus perpetuating the sepsis cascade.

[0370] Stage 5 is characterized by MODS, accompanied by persistent dysregulation of SIRS and CARS responses.

[0371] In other embodiments, the acute inflammatory condition is associated with acute respiratory distress syndrome (ARDS). In an exemplary embodiment, the subject may have one or more symptoms selected from the following: mild, moderate, or severe hypoxemia (determined by arterial oxygen partial pressure / inspiratory oxygen concentration (PaO2 / FiO2) or positive end-expiratory pressure (PEEP), bilateral lung shadows, respiratory failure, shortness of breath, dyspnea, cough, fever, tachycardia, hypotension, confusion, extreme fatigue, rapid breathing, organ failure, chest pain, cyanosis of nails or lips, changes in the levels of one or more inflammatory markers, or the need for mechanical ventilation.

[0372] In some implementations, the acute inflammatory condition is associated with severe acute respiratory syndrome (SARS), wherein, for example, the subject may have one or more symptoms selected from the following: acute febrile illness, weakness, fatigue, headache, flushing, diarrhea, nausea, vomiting, cough (including dry cough), sore throat, runny nose, nasal congestion, production of pro-inflammatory mediators, vascular leakage, and organ failure.

[0373] The present invention further provides a method for treating cytokine release syndrome (CRS) or cytokine storm in a subject, wherein the CRS or cytokine storm is associated with coronavirus infection, the method comprising, consisting of, or substantially consisting of administering to the subject a compound of formula I, II, III, IV, V, VI, or VII or a pharmaceutically acceptable salt, solvate, or prodrug thereof. Further provided are the use of compounds of formula I, II, III, IV, V, VI, or VII, or pharmaceutically acceptable salts, solvates, or prodrugs thereof, for treating cytokine release syndrome (CRS) or cytokine storm in subjects, wherein the CRS or cytokine storm is associated with coronavirus infection; and the use of compounds of formula I, II, III, IV, V, VI, or VII, or pharmaceutically acceptable salts, solvates, or prodrugs thereof, for treating cytokine release syndrome (CRS) or cytokine storm in subjects, wherein the CRS or cytokine storm is associated with coronavirus infection; and the use of compounds of formula I, II, III, IV, V, VI, or VII, or pharmaceutically acceptable salts, solvates, or prodrugs thereof, in the preparation of a medicament for treating cytokine release syndrome (CRS) or cytokine storm in subjects, wherein the CRS or cytokine storm is associated with coronavirus infection.

[0374] Suitable embodiments of compounds of formula I, II, III, IV, V, VI, or VII have been discussed above.

[0375] In some embodiments, the compound is a compound of formula I, II, or III, or a pharmaceutically acceptable salt, solvate, or prodrug thereof; particularly a compound of formula III, or a pharmaceutically acceptable salt, solvate, or prodrug thereof. In some embodiments, the compound is a compound of formula I, II, or III, or a pharmaceutically acceptable salt thereof; particularly a compound of formula III, or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is a compound of formula I, II, or III; particularly a compound of formula III.

[0376] In some embodiments, the compound is a compound of formula IV, V, or VI, or a pharmaceutically acceptable salt, solvate, or prodrug thereof; particularly a compound of formula VI, or a pharmaceutically acceptable salt, solvate, or prodrug thereof. In some embodiments, the compound is a compound of formula IV, V, or VI, or a pharmaceutically acceptable salt thereof; particularly a compound of formula VI, or a pharmaceutically acceptable salt thereof. In specific embodiments, the compound is a compound of formula IV, V, or VI; particularly a compound of formula VI.

[0377] In some embodiments, the coronavirus is a beta-coronavirus, such as lineage A beta-coronavirus, lineage B beta-coronavirus, lineage C beta-coronavirus, or lineage D beta-coronavirus. In some embodiments, the beta-coronavirus is lineage B beta-coronavirus, such as SARS-CoV or SARS-CoV-2; particularly SARS-CoV-2. In alternative embodiments, the beta-coronavirus is lineage C beta-coronavirus, such as MERS-CoV. In specific embodiments, the coronavirus is an ACE2-binding coronavirus, such as lineage B beta-coronavirus, including SARS-CoV or SARS-CoV-2.

[0378] In a particular embodiment, the CRS or cytokine storm comprises an increase of at least 50% compared to the baseline state of one or more cytokines or chemokines selected from IFN-γ, IFN-β, TNF-α, IL-1β, IL-6, IL-17A, CCL3, CXCL2, and CXCL8.

[0379] Symptoms associated with CRS include, but are not limited to, one or more of the following: fever, fatigue, loss of appetite, muscle and joint pain, nausea, vomiting, diarrhea, rash, rapid breathing, rapid heartbeat, low blood pressure, seizures, headache, confusion, delirium, hallucinations, tremor, and coordination disorder.

[0380] Non-restrictive symptoms associated with cytokine storm include one or more of the following: high fever, swelling and redness, extreme fatigue, nausea, bleeding, clotting, visceral organ damage, and shock, or any combination thereof.

[0381] The CRS or cytokine storm can be associated with MIS-C. In such an implementation, the subject may have one or more symptoms selected from the following: fever, vomiting, diarrhea, stomach pain, rash, red eyes, red or swollen lips and tongue, unusual fatigue, red or swollen hands or feet, severe stomach pain, cardiac symptoms (including chest pain, palpitations, and shortness of breath), cyanosis of lips or face, confusion, inability to be awakened or remain conscious, abdominal pain with vomiting and diarrhea, rash and swelling of the extremities, syncope, and new-onset hypotension.

[0382] In some implementations, the CRS or cytokine storm is associated with SIRS, such as stage 1 SIRS, stage 2 SIRS, stage 3 SIRS, stage 4 SIRS, or stage 5 SIRS.

[0383] In some implementations, the CRS or cytokine storm is associated with ARDS. For example, the subject may have one or more symptoms selected from the following: mild, moderate, or severe hypoxemia (determined by arterial oxygen partial pressure / inspiratory oxygen concentration (PaO2 / FiO2) or positive end-expiratory pressure (PEEP), bilateral lung shadows, respiratory failure, shortness of breath, dyspnea, cough, fever, tachycardia, hypotension, confusion, extreme fatigue, rapid breathing, organ failure, chest pain, cyanosis of nails or lips, changes in the levels of one or more inflammatory markers, or the need for mechanical ventilation.

[0384] In a particular implementation, the CRS or cytokine storm is associated with SARS. For example, the subject may have one or more symptoms selected from the following: acute febrile illness, weakness, fatigue, headache, flushing, diarrhea, nausea, vomiting, cough (including dry cough), sore throat, runny nose, nasal congestion, production of pro-inflammatory mediators, vascular leakage, and organ failure.

[0385] Treatment of SARS is also considered. Therefore, another aspect of the invention provides a method for treating SARS in a subject, wherein the SARS is associated with coronavirus infection, the method comprising, consisting of, or substantially consisting of: administering to the subject a compound of formula I, II, III, IV, V, VI, or VII, or a pharmaceutically acceptable salt, solvate, or prodrug thereof. This document also provides the use of a compound of formula I, II, III, IV, V, VI, or VII, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, for treating SARS in a subject, wherein the SARS is associated with coronavirus infection; the use of a compound of formula I, II, III, IV, V, VI, or VII, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, for treating SARS in a subject, wherein the SARS is associated with coronavirus infection; and the use of a compound of formula I, II, III, IV, V, VI, or VII, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, in the preparation of a medicament for treating SARS in a subject, wherein the SARS is associated with coronavirus infection.

[0386] Suitable embodiments of compounds of formulas I, II, III, IV, V, VI, and VII have been discussed above.

[0387] In some embodiments, the coronavirus is a beta-coronavirus, such as lineage A beta-coronavirus, lineage B beta-coronavirus, lineage C beta-coronavirus, or lineage D beta-coronavirus. In some embodiments, the beta-coronavirus is lineage B beta-coronavirus, such as SARS-CoV or SARS-CoV-2; particularly SARS-CoV-2. In alternative embodiments, the beta-coronavirus is lineage C beta-coronavirus, such as MERS-CoV. In specific embodiments, the coronavirus is an ACE2-binding coronavirus, such as lineage B beta-coronavirus, including SARS-CoV or SARS-CoV-2.

[0388] The subject may have, for example, one or more of the following symptoms: acute febrile illness, weakness, fatigue, headache, flushing, diarrhea, nausea, vomiting, cough (including dry cough), sore throat, runny nose, nasal congestion, production of pro-inflammatory mediators, vascular leakage and organ failure, or any combination thereof.

[0389] In some embodiments, the SARS is associated with the presence of CRS or a cytokine storm, for example, wherein the CRS or cytokine storm comprises an increase of at least 50% compared to the baseline state of one or more cytokines or chemokines selected from IFN-γ, IFN-β, TNF-α, IL-1β, IL-6, IL-17A, CCL3, CXCL2, and CXCL8.

[0390] The SARS mentioned can also be associated with MISC-C, wherein, for example, the subject may have one or more of the following symptoms: fever, vomiting, diarrhea, stomach pain, rash, red eyes, red or swollen lips and tongue, unusual fatigue, red or swollen hands or feet, severe stomach pain, cardiac symptoms (including chest pain, palpitations and shortness of breath), bluish lips or face, confusion, inability to be awakened or remain conscious, abdominal pain with vomiting and diarrhea, rash and swelling of the extremities, syncope and new-onset hypotension.

[0391] The SARS mentioned can also be associated with SIRS. For example, the subjects may have stage 1 SIRS, stage 2 SIRS, stage 3 SIRS, stage 4 SIRS, or stage 5 SIRS.

[0392] In another aspect, a method for antagonizing a viral fusion protein is provided, the method comprising contacting a virus expressing the viral fusion protein with a compound of the present invention. Further consideration is given to the use of compounds of formula I, II, III, IV, V, VI, or VII for antagonizing viral fusion proteins, or pharmaceutically acceptable salts, solvates, or prodrugs thereof, as well as the use of compounds of formula I, II, III, IV, V, VI, or VII, or pharmaceutically acceptable salts, solvates, or prodrugs thereof, for antagonizing viral fusion proteins.

[0393] Preferably, the viral fusion protein is antagonized in the subject, thereby treating or inhibiting the development of viral infection. Therefore, a method for antagonizing a viral fusion protein in a subject is also provided, the method comprising administering to the subject a compound of formula I, II, III, IV, V, VI, or VII, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein the compound contacts a virus expressing the fusion protein and antagonizes the fusion protein. In another aspect, there are provided the use of a compound of formula I, II, III, IV, V, VI, or VII, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, for antagonizing a viral fusion protein in a subject, wherein said compound contacts a virus expressing the fusion protein and antagonizes the fusion protein; the use of a compound of formula I, II, III, IV, V, VI, or VII, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, for antagonizing a viral fusion protein in a subject, wherein said compound contacts a virus expressing the fusion protein and antagonizes the fusion protein; and the use of a compound of formula I, II, III, IV, V, VI, or VII, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, in the preparation of a medicament for antagonizing a viral fusion protein in a subject, wherein said compound contacts a virus expressing the fusion protein and antagonizes the fusion protein.

[0394] The above discussion covered suitable implementation schemes for viruses and compounds.

[0395] In some embodiments, the viral fusion protein is a spike protein.

[0396] As discussed herein, in certain embodiments, the virus is a coronavirus, such as SARS-CoV, SARS-CoV-2, or MERS; particularly SARS-CoV-2. Therefore, in certain embodiments, the viral fusion protein is a coronavirus spike protein, such as the SARS-CoV spike protein, the SARS-CoV-2 spike protein, or the MERS spike protein; particularly the SARS-CoV or SARS-CoV-2 spike protein; most particularly the SARS-CoV-2 spike protein.

[0397] Antagonizing viral fusion proteins inhibits at least one activity of the fusion protein. In a particular embodiment, at least one activity of the viral fusion protein is either viral binding to a host cell or viral entry into the cell; preferably, both viral binding to a host cell and viral entry into the cell. Suitable host cells include lung cells (e.g., alveolar cells), intestinal cells, endothelial cells, epithelial cells (e.g., nasal or nasopharyngeal epithelial cells), kidney cells (e.g., brush border cells of proximal renal tubular epithelial cells), or arterial smooth muscle cells. In a particular embodiment, the cells are respiratory cells. In a preferred embodiment, the cells are cells expressing the ACE2 peptide.

[0398] When administered to any subject in any of the foregoing aspects, the subject has or is suspected of having a viral infection, or has been exposed to or is at risk of exposure to a virus capable of causing a viral infection. In a particular embodiment, the subject is a human being.

[0399] To avoid being bound by theory, the compounds of this invention bind to fusion proteins and directly inhibit interactions with host cells (particularly surface proteins on host cells), for example, by binding to fusion proteins at surface protein binding sites; or by binding to fusion proteins and causing conformational changes in the fusion proteins, thereby inhibiting interactions with host cells (e.g., conformational changes inhibit the binding of fusion proteins to surface proteins on host cells). Therefore, while the compounds may bind (completely or partially) to fusion proteins at host cell surface protein binding sites, they may alternatively bind to fusion proteins at sites other than the host cell surface protein binding sites.

[0400] While the administration of a single compound is considered, the present invention also contemplates administering to the subject, in any of the foregoing aspects, a second compound of formula I, II, III, IV, V, VI, or VII, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, thereof. The second compound may be administered in parallel with the first compound, for example, in a single composition. It is not intended to be theoretically restrictive, but it is thought that the administration of two different compounds of the present invention may enhance the inhibition of viral fusion proteins. For example, it is suggested to administer in parallel a compound that binds directly to a host cell surface protein binding site of the viral fusion protein and a compound that binds to a different site causing a conformational change in the viral fusion protein. In some embodiments, the method comprises administering a compound of formula II, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, and a compound of formula III, or a pharmaceutically acceptable salt, solvate, or prodrug thereof. In a particular embodiment, the method comprises administering a compound of formula V, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, and a compound of formula VI, or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

[0401] Any of the foregoing aspects may relate to administering an effective amount of the compound of the invention, as described in Part 3 above. The compound may be administered in the form of, for example, the pharmaceutical compositions described herein. The compounds of the invention may be administered to a subject via any route of administration suitable for treating or inhibiting the development of a viral infection, treating a condition, or antagonizing a fusion protein. Those skilled in the art will readily recognize suitable routes of administration, such as one or more discussed in Part 3 above. For example, in some embodiments, the compound is administered orally, by inhalation, intranasal administration, topical administration, or intravenous administration; particularly by inhalation or intranasal administration.

[0402] The dosage and frequency of administration will depend on the subject, the disorder to be treated, and the route of administration. A technician will be able to readily determine the appropriate dosage and the frequency of such dosage. For example, the compound can be administered in amounts ranging from about 0.25 μg to about 2000 mg, and can be administered, for example, once daily, twice daily, or three times daily. The treatment may last for days, weeks, months, or years. In embodiments where the pharmaceutical composition comprises one or more additional active agents, the dosage and frequency of administration are determined with reference to the commonly used dosage and route of administration of the pharmaceutical agent.

[0403] In some implementations, any of the above methods may involve the application of one or more other active agents described in Part 3 above, such as antimicrobial agents (e.g., antiviral agents).

[0404] Technicians will be well aware of the appropriate assays for evaluating the antagonistic effects of viral fusion proteins. For example, the method may include contacting a virus or cell expressing a viral fusion protein (e.g., spike protein) with a compound and screening for inhibition of binding of the fusion protein to its partner (such as ACE2) (e.g., ligand binding assay). Alternatively, the method may include screening for inhibition of the activity, presence, or expression of downstream cellular targets or products (e.g., viral entry into host cells or the presence of viral infection in a subject (e.g., an animal model)). Detection of such binding, activity, presence, or expression can be achieved using a variety of techniques, including, but not limited to, ELISA, ligand binding assays (e.g., radioligand binding assays or fluorescence binding assays), surface plasmon resonance, immunofluorescence, Western blotting, immunoprecipitation, immunostaining, scintillation proximity assays, or cell proliferation assays. Commercially available kits and / or products may also be used, such as the SARS-CoV-2 spike-ACE2 interaction inhibitor screening assay kit (catalog number 502050; Cayman Chemical, Ann Arbor, Michigan, USA); or the spike S1 (SARS-CoV-2):ACE2 inhibitor screening colorimetric assay kit (catalog number 79954; BPSBioscience, San Diego, California, USA).

[0405] Standard techniques can also be used to assess the interaction between the virus and cells expressing ACE2, such as binding assays discussed above (including ligand binding assays, ELISA, surface plasmon resonance, etc.), which detect the interaction between viral proteins (e.g., spike proteins) and ACE2, or assays such as immunostaining can be used to determine viral infection in host cells. Animal models can also be used to assess viral infection.

[0406] Implementation Plan

[0407] Exemplary implementations include, but are not limited to:

[0408] 1. Compounds of formula I, II, or III:

[0409]

[0410] (I)

[0411]

[0412] (II)

[0413]

[0414] (III)

[0415] Or its pharmaceutically acceptable salt, solvate, or prodrug,

[0416] in:

[0417] L 1 L 3 L 4 L 6 L 7 L 8 L 9 L 10 L 11 L 12 L 13 L 15 L 16 L 17 L 18 and L 19 Independently selected from optionally substituted C1-C5 alkylene groups, optionally substituted C2-C5 alkenyl groups, and optionally substituted C2-C5 ynylene groups;

[0418] L 2 L 5 and L 14 Independently selected from optionally substituted C2-C8 alkylene groups, optionally substituted C2-C8 alkenyl groups, and optionally substituted C2-C8 ynylene groups;

[0419] X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 and X 11 Independently selected from CH and N; and

[0420] R 1 Selected from C2-C which are optionally replaced 12 Alkyl groups, optionally substituted C2-C 12 Alkenyl and optionally substituted C2-C 12 Alkyne group.

[0421] 2. The compound according to embodiment 1, wherein X 1 -X 11 It is CH.

[0422] 3. The compound according to embodiment 1 or embodiment 2, wherein L 1 L 3 L 4 L 6 L7 L 8 L 9 L 10 L 11 L 12 L 13 L 15 L 16 L 17 L 18 and L 19 Independently selected from optionally substituted C1-C3 alkylene groups, optionally substituted C2-C3 alkenyl groups, and optionally substituted C2-C3 ynylene groups.

[0423] 4. The compound according to embodiment 3, wherein L 1 L 3 L 4 L 6 L 7 L 8 L 9 L 10 L 11 L 12 L 13 L 15 L 16 L 17 L 18 and L 19 It is a C1-C3 alkylene group that is optionally substituted.

[0424] 5. The compound according to embodiment 4, wherein L 1 L 3 L 4 L 6 L 7 L 8 L 9 L 10 L 11 L 12 L 13 L 15 L 16 L 17 L 18 and L 19 It is methylene.

[0425] 6. A compound according to any one of embodiments 1-5, wherein L 2 It is selected from optionally substituted C2-C5 alkylene groups, optionally substituted C2-C5 alkenyl groups, and optionally substituted C2-C5 ynylene groups.

[0426] 7. The compound according to embodiment 6, wherein L 2It is an optional substituted C2-C5 alkylene group.

[0427] 8. The compound according to embodiment 7, wherein L 2 It is propylidene.

[0428] 9. A compound according to any one of embodiments 1-8, wherein L 5 It is selected from optionally substituted C3-C6 alkylene groups, optionally substituted C3-C6 alkenyl groups, and optionally substituted C6-C6 alkyne groups.

[0429] 10. The compound according to embodiment 9, wherein L 5 It is a C3-C6 alkylene group that is optionally substituted.

[0430] 11. The compound according to embodiment 10, wherein L 5 It is Adinaki.

[0431] 12. A compound according to any one of embodiments 1-11, wherein L 14 It is selected from optionally substituted C2-C4 alkylene groups, optionally substituted C2-C4 alkenyl groups, and optionally substituted C2-C4 ynylene groups.

[0432] 13. The compound according to embodiment 12, wherein L 14 It is a C2-C4 alkylene group that is optionally substituted.

[0433] 14. The compound according to embodiment 13, wherein L 14 It is ethylene.

[0434] 15. A compound according to any one of embodiments 1-14, wherein R 1 Selected from C6-C which are optionally replaced 10 Alkyl groups, optionally substituted C6-C 10 Alkenyl and optionally substituted C6-C 10 Alkyne group.

[0435] 16. The compound according to embodiment 15, wherein R 1 C6-C is arbitrarily replaced. 10 alkyl.

[0436] 17. The compound according to embodiment 16, wherein R 1 It's Hsinki.

[0437] 18. A compound according to any one of embodiments 1-17, wherein said compound is a compound of formula IV:

[0438]

[0439] (IV)

[0440] Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0441] 19. A compound according to any one of embodiments 1-17, wherein said compound is a compound of formula V:

[0442]

[0443] (V)

[0444] Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0445] 20. A compound according to any one of embodiments 1-17, wherein said compound is a compound of formula VI:

[0446]

[0447] (VI)

[0448] Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0449] 21. A compound according to any one of embodiments 1-20, wherein the compound is in the form of a salt and the salt is a sodium salt.

[0450] 22. A pharmaceutical composition comprising a compound according to any one of embodiments 1-21, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, and a pharmaceutically acceptable carrier or diluent.

[0451] 23. A compound of any one of embodiments 1-21, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, used in a therapy.

[0452] 24. A method for treating or inhibiting the development of a viral infection in a subject, said method comprising, consisting of, or substantially consisting of: administering to the subject a compound according to any one of embodiments 1-21 or a pharmaceutically acceptable salt, solvate, or prodrug thereof, or a compound of formula VII or a pharmaceutically acceptable salt, solvate, or prodrug thereof:

[0453]

[0454] (VII)

[0455] 25. The method according to embodiment 24, wherein the infection is caused by a virus expressing a viral fusion protein.

[0456] 26. The method according to implementation scheme 24 or implementation scheme 25, wherein the infection is caused by an enveloped virus.

[0457] 27. The method according to any one of embodiments 24-26, wherein the infection is caused by a virus interacting with a cell expressing the ACE2 polypeptide.

[0458] 28. The method according to any one of embodiments 24-27, wherein the viral infection is a coronavirus infection.

[0459] 29. The method according to implementation plan 28, wherein the coronavirus is capable of causing severe acute respiratory syndrome (SARS).

[0460] 30. The method according to implementation scheme 28 or implementation scheme 29, wherein the coronavirus is a beta-coronavirus.

[0461] 31. The method according to embodiment 30, wherein the beta-coronavirus is selected from lineage A beta-coronavirus, lineage B beta-coronavirus, lineage C beta-coronavirus and lineage D beta-coronavirus.

[0462] 32. The method according to implementation scheme 31, wherein the beta-coronavirus is lineage B beta-coronavirus.

[0463] 33. The method according to implementation scheme 32, wherein the lineage B beta-coronavirus is selected from SARS-CoV and SARS-CoV-2.

[0464] 34. The method according to implementation scheme 33, wherein the lineage B beta-coronavirus is SARS-CoV-2.

[0465] 35. The method according to implementation scheme 31, wherein the beta coronavirus is lineage C beta coronavirus.

[0466] 36. The method according to implementation scheme 35, wherein the lineage C beta coronavirus is MERS-CoV.

[0467] 37. The method according to any one of embodiments 24-36, wherein the compound is administered orally, by inhalation, intranasally, topically or intravenously.

[0468] 38. The method according to any one of embodiments 24-37, wherein the method further comprises administering a second compound of formula I, II, III, IV, V, VI or VII, or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0469] 39. The method according to embodiment 38, wherein the method comprises administering a compound of formula V or a pharmaceutically acceptable salt, solvate or prodrug thereof and a compound of formula VI or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0470] 40. A method for inhibiting the interaction between a virus and cells expressing an ACE2 polypeptide, the method comprising, consisting of, or substantially consisting of: contacting the virus with a compound according to any one of embodiments 1-21 or a pharmaceutically acceptable salt, solvate, or prodrug thereof, or a compound of formula VII or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

[0471] 41. The method according to embodiment 40, wherein the interaction is one or both of virus binding to a cell and virus entry into a cell.

[0472] 42. The method according to embodiment 40 or embodiment 41, wherein the cells are lung cells (e.g., alveolar cells), intestinal cells, endothelial cells, epithelial cells (e.g., nasal or nasopharyngeal epithelial cells), kidney cells (e.g., brush border of proximal renal tubular epithelial cells) or arterial smooth muscle cells.

[0473] 43. The method according to any one of embodiments 40-42, wherein the cells are respiratory cells.

[0474] 44. The method according to any one of embodiments 40-43, wherein the virus is a coronavirus.

[0475] 45. The method according to implementation scheme 44, wherein the coronavirus is capable of causing SARS.

[0476] 46. ​​The method according to implementation scheme 44 or implementation scheme 45, wherein the coronavirus is a beta-coronavirus.

[0477] 47. The method according to implementation scheme 46, wherein the betacoronavirus is selected from lineage A betacoronavirus, lineage B betacoronavirus, lineage C betacoronavirus and lineage D betacoronavirus.

[0478] 48. The method according to implementation scheme 47, wherein the beta-coronavirus is lineage B beta-coronavirus.

[0479] 49. The method according to implementation scheme 48, wherein the lineage B beta-coronavirus is selected from SARS-CoV and SARS-CoV-2.

[0480] 50. The method according to implementation scheme 49, wherein the lineage B beta-coronavirus is SARS-CoV-2.

[0481] 51. The method according to implementation scheme 47, wherein the beta-coronavirus is lineage C beta-coronavirus.

[0482] 52. The method according to implementation scheme 51, wherein the lineage C beta coronavirus is MERS-CoV.

[0483] 53. The method according to any one of embodiments 40-52, wherein the compound is administered orally, by inhalation, intranasally, topically or intravenously.

[0484] 54. The method according to any one of embodiments 40-53, wherein the method further comprises administering a second compound of formula I, II, III, IV, V, VI or VII, or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0485] 55. The method according to embodiment 54, wherein the method comprises administering a compound of formula V or a pharmaceutically acceptable salt, solvate or prodrug thereof and a compound of formula VI or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0486] 56. A method for treating an acute inflammatory condition in a subject, wherein the condition is associated with a coronavirus infection, the method comprising, consisting of, or substantially consisting of: administering to the subject a compound according to any one of embodiments 1-21 or a pharmaceutically acceptable salt, solvate, or prodrug thereof, or a compound of formula VII or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

[0487] 57. The method according to implementation scheme 56, wherein the coronavirus is a beta-coronavirus.

[0488] 58. The method according to implementation scheme 57, wherein the betacoronavirus is selected from lineage A betacoronavirus, lineage B betacoronavirus, lineage C betacoronavirus and lineage D betacoronavirus.

[0489] 59. The method according to implementation scheme 58, wherein the beta coronavirus is lineage B beta coronavirus.

[0490] 60. The method according to embodiment 59, wherein the lineage B beta-coronavirus is selected from SARS-CoV and SARS-CoV-2.

[0491] 61. The method according to implementation scheme 60, wherein the lineage B beta-coronavirus is SARS-CoV-2.

[0492] 62. The method according to implementation scheme 58, wherein the beta-coronavirus is lineage C beta-coronavirus.

[0493] 63. The method according to implementation scheme 62, wherein the lineage C beta coronavirus is MERS-CoV.

[0494] 64. The method according to any one of embodiments 56-63, wherein the acute inflammatory condition is associated with the presence of cytokine release syndrome (CRS) or cytokine storm.

[0495] 65. The method according to embodiment 64, wherein the CRS or cytokine storm comprises an increase of at least 50% compared to the baseline state of one or more cytokines or chemokines selected from IFN-γ, IFN-β, TNF-α, IL-1β, IL-6, IL-17A, CCL3, CXCL2, and CXCL8.

[0496] 66. The method according to implementation scheme 64 or implementation scheme 65, wherein the subject has CRS and has one or more symptoms selected from the following: fever, fatigue, loss of appetite, muscle and joint pain, nausea, vomiting, diarrhea, rash, rapid breathing, rapid heartbeat, hypotension, seizures, headache, confusion, delirium, hallucinations, tremor and coordination disorder.

[0497] 67. The method according to embodiment 64 or embodiment 65, wherein the subject has a cytokine storm and has one or more symptoms selected from: high fever, swelling and redness, extreme fatigue, nausea, bleeding, coagulation, visceral organ damage and shock or any combination thereof.

[0498] 68. The method according to any one of embodiments 56-67, wherein the acute inflammatory condition is associated with multisystem inflammatory syndrome (MIS-C) in children.

[0499] 69. The method according to implementation scheme 68, wherein the subject has one or more of the following symptoms: fever, vomiting, diarrhea, stomach pain, rash, red eyes, red or swollen lips and tongue, unusual fatigue, red or swollen hands or feet, severe stomach pain, cardiac symptoms (including chest pain, palpitations and shortness of breath), cyanosis of lips or face, confusion, inability to be awakened or remain conscious, abdominal pain with vomiting and diarrhea, rash and swelling of the extremities, syncope and new-onset hypotension.

[0500] 70. The method according to any one of embodiments 56-59, wherein the acute inflammatory condition is associated with systemic inflammatory response syndrome (SIRS).

[0501] 71. The method according to embodiment 70, wherein the subject has stage 1 SIRS.

[0502] 72. The method according to embodiment 70, wherein the subject has stage 2 SIRS.

[0503] 73. The method according to embodiment 70, wherein the subject has stage 3 SIRS.

[0504] 74. The method according to embodiment 70, wherein the subject has stage 4 SIRS.

[0505] 75. The method according to embodiment 70, wherein the subject has stage 5 SIRS.

[0506] 76. The method according to any one of embodiments 56-69, wherein the acute inflammatory condition is associated with acute respiratory distress syndrome (ARDS).

[0507] 77. The method according to embodiment 76, wherein the subject has one or more of the following symptoms: mild, moderate or severe hypoxemia (determined by arterial oxygen partial pressure / inspiratory oxygen concentration (PaO2 / FiO2) or positive end-expiratory pressure (PEEP), bilateral lung shadows, respiratory failure, shortness of breath, dyspnea, cough, fever, tachycardia, hypotension, confusion, extreme fatigue, rapid breathing, organ failure, chest pain, cyanosis of nails or lips, changes in the levels of one or more inflammatory markers, or the need for mechanical ventilation.

[0508] 78. The method according to any one of embodiments 56-69, wherein the acute inflammatory condition is associated with SARS.

[0509] 79. The method according to implementation scheme 78, wherein the subject has one or more symptoms selected from the following: acute febrile illness, weakness, fatigue, headache, flushing, diarrhea, nausea, vomiting, cough (including dry cough), sore throat, runny nose, nasal congestion, production of pro-inflammatory mediators, vascular leakage, and organ failure.

[0510] 80. A method for treating cytokine release syndrome (CRS) or cytokine storm in a subject, wherein the CRS or cytokine storm is associated with coronavirus infection, the method comprising, consisting of, or substantially consisting of: administering to the subject a compound according to any one of embodiments 1-21 or a pharmaceutically acceptable salt, solvate, or prodrug thereof, or a compound of formula VII or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

[0511] 81. The method according to implementation scheme 80, wherein the coronavirus is a beta-coronavirus.

[0512] 82. The method according to implementation scheme 81, wherein the beta-coronavirus is selected from lineage A beta-coronavirus, lineage B beta-coronavirus, lineage C beta-coronavirus and lineage D beta-coronavirus.

[0513] 83. The method according to implementation scheme 82, wherein the beta-coronavirus is lineage B beta-coronavirus.

[0514] 84. The method according to implementation scheme 83, wherein the lineage B beta-coronavirus is selected from SARS-CoV and SARS-CoV-2.

[0515] 85. The method according to implementation scheme 84, wherein the lineage B beta-coronavirus is SARS-CoV-2.

[0516] 86. The method according to implementation scheme 82, wherein the beta-coronavirus is lineage C beta-coronavirus.

[0517] 87. The method according to implementation scheme 86, wherein the lineage of beta-coronavirus C is MERS-CoV.

[0518] 88. The method according to any one of embodiments 80-87, wherein the CRS or cytokine storm comprises an increase of at least 50% compared to the baseline state of one or more cytokines or chemokines selected from IFN-γ, IFN-β, TNF-α, IL-1β, IL-6, IL-17A, CCL3, CXCL2 and CXCL8.

[0519] 89. The method according to any one of embodiments 80-88, wherein the subject has CRS and has one or more symptoms selected from the following: fever, fatigue, loss of appetite, muscle and joint pain, nausea, vomiting, diarrhea, rash, rapid breathing, rapid heartbeat, hypotension, seizures, headache, confusion, delirium, hallucinations, tremor and coordination disorder.

[0520] 90. The method according to any one of embodiments 80-89, wherein the subject has a cytokine storm and has one or more symptoms selected from: high fever, swelling and redness, extreme fatigue, nausea, bleeding, coagulation, visceral organ damage and shock or any combination thereof.

[0521] 91. The method according to any one of embodiments 80-90, wherein the CRS or cytokine storm is associated with MIS-C.

[0522] 92. The method of implementation scheme 91, wherein the subject has one or more symptoms selected from the following: fever, vomiting, diarrhea, stomach pain, rash, red eyes, red or swollen lips and tongue, unusual fatigue, red or swollen hands or feet, severe stomach pain, cardiac symptoms (including chest pain, palpitations and shortness of breath), cyanosis of lips or face, confusion, inability to be awakened or remain conscious, abdominal pain with vomiting and diarrhea, rash and swelling of the extremities, syncope and new-onset hypotension.

[0523] 93. The method according to any one of embodiments 80-90, wherein the CRS or cytokine storm is associated with SIRS.

[0524] 94. The method according to embodiment 93, wherein the subject has stage 1 SIRS.

[0525] 95. The method according to embodiment 93, wherein the subject has stage 2 SIRS.

[0526] 96. The method according to embodiment 93, wherein the subject has stage 3 SIRS.

[0527] 97. The method according to embodiment 93, wherein the subject has stage 4 SIRS.

[0528] 98. The method according to embodiment 93, wherein the subject has stage 5 SIRS.

[0529] 99. The method according to any one of embodiments 80-90, wherein the CRS or cytokine storm is associated with ARDS.

[0530] 100. The method according to embodiment 99, wherein the subject has one or more symptoms selected from the following: mild, moderate or severe hypoxemia (determined by arterial oxygen partial pressure / inspiratory oxygen concentration (PaO2 / FiO2) or positive end-expiratory pressure (PEEP), bilateral lung shadows, respiratory failure, shortness of breath, dyspnea, cough, fever, tachycardia, hypotension, confusion, extreme fatigue, rapid breathing, organ failure, chest pain, cyanosis of nails or lips, changes in the levels of one or more inflammatory markers, or the need for mechanical ventilation.

[0531] 101. The method according to any one of embodiments 80-90, wherein the CRS or cytokine storm is associated with SARS.

[0532] 102. The method according to implementation scheme 101, wherein the subject has one or more symptoms selected from the following: acute febrile illness, weakness, fatigue, headache, flushing, diarrhea, nausea, vomiting, cough (including dry cough), sore throat, runny nose, nasal congestion, production of pro-inflammatory mediators, vascular leakage and organ failure.

[0533] 103. A method for treating a subject with SARS, wherein the SARS is associated with coronavirus infection, the method comprising, consisting of, or substantially consisting of: administering to the subject a compound according to any one of embodiments 1-21 or a pharmaceutically acceptable salt, solvate, or prodrug thereof, or a compound of formula VII or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

[0534] 104. The method according to implementation scheme 103, wherein the coronavirus is a beta-coronavirus.

[0535] 105. The method according to embodiment 104, wherein the beta-coronavirus is selected from lineage A beta-coronavirus, lineage B beta-coronavirus, lineage C beta-coronavirus and lineage D beta-coronavirus.

[0536] 106. The method according to implementation scheme 105, wherein the beta coronavirus is lineage B beta coronavirus.

[0537] 107. The method according to embodiment 106, wherein the lineage B beta-coronavirus is selected from SARS-CoV and SARS-CoV-2.

[0538] 108. The method according to implementation scheme 107, wherein the lineage B beta-coronavirus is SARS-CoV-2.

[0539] 109. The method according to implementation scheme 105, wherein the beta coronavirus is lineage C beta coronavirus.

[0540] 110. The method according to implementation scheme 109, wherein the lineage C beta coronavirus is MERS-CoV.

[0541] 111. The method according to any one of embodiments 103-110, wherein the subject has one or more symptoms selected from: acute febrile illness, weakness, fatigue, headache, flushing, diarrhea, nausea, vomiting, cough (including dry cough), sore throat, runny nose, nasal congestion, production of pro-inflammatory mediators, vascular leakage and organ failure or any combination thereof.

[0542] 112. The method according to any one of embodiments 103-111, wherein the SARS is associated with the presence of CRS or cytokine storm.

[0543] 113. The method according to embodiment 112, wherein the CRS or cytokine storm comprises an increase of at least 50% compared to the baseline state of one or more cytokines or chemokines selected from IFN-γ, IFN-β, TNF-α, IL-1β, IL-6, IL-17A, CCL3, CXCL2, and CXCL8.

[0544] 114. The method according to any one of implementation schemes 103-113, wherein the SARS is associated with MISC-C.

[0545] 115. The method according to embodiment 114, wherein the subject has one or more symptoms selected from the following: fever, vomiting, diarrhea, stomach pain, rash, red eyes, red or swollen lips and tongue, unusual fatigue, red or swollen hands or feet, severe stomach pain, cardiac symptoms (including chest pain, palpitations and shortness of breath), cyanosis of lips or face, confusion, inability to be awakened or remain conscious, abdominal pain with vomiting and diarrhea, rash and swelling of the extremities, syncope and new-onset hypotension.

[0546] 116. The method according to any one of implementation schemes 103-115, wherein the SARS is associated with SIRS.

[0547] 117. The method according to embodiment 116, wherein the subject has stage 1 SIRS.

[0548] 118. The method according to embodiment 116, wherein the subject has stage 2 SIRS.

[0549] 119. The method according to embodiment 104, wherein the subject has stage 3 SIRS.

[0550] 120. The method according to embodiment 116, wherein the subject has stage 4 SIRS.

[0551] 121. The method according to implementation scheme 116, wherein the subject has stage 5 SIRS.

[0552] 122. The use of a compound of any one of embodiments 1-21 or a pharmaceutically acceptable salt, solvate or prodrug thereof, or a compound of formula VII or a pharmaceutically acceptable salt, solvate or prodrug thereof, in the preparation of a medicament for treating or inhibiting the development of a viral infection in a subject.

[0553] 123. The use of a compound of any one of embodiments 1-21 or a pharmaceutically acceptable salt, solvate or prodrug thereof, or a compound of formula VII or a pharmaceutically acceptable salt, solvate or prodrug thereof, in the preparation of a medicament for treating an acute inflammatory condition in a subject, wherein the condition is associated with coronavirus infection.

[0554] 124. The use of a compound of any one of embodiments 1-21 or a pharmaceutically acceptable salt, solvate or prodrug thereof, or a compound of formula VII or a pharmaceutically acceptable salt, solvate or prodrug thereof, in the preparation of a medicament for treating a subject with CRS or cytokine storm, wherein the CRS or cytokine storm is associated with coronavirus infection.

[0555] 125. The use of a compound of any one of embodiments 1-21 or a pharmaceutically acceptable salt, solvate or prodrug thereof, or a compound of formula VII or a pharmaceutically acceptable salt, solvate or prodrug thereof, in the preparation of a medicament for treating SARS in a subject, wherein the SARS is associated with coronavirus infection.

[0556] 126. A compound of any one of embodiments 1-21 or a pharmaceutically acceptable salt, solvate or prodrug thereof, or a compound of formula VII or a pharmaceutically acceptable salt, solvate or prodrug thereof, used to treat or inhibit the development of a viral infection in a subject.

[0557] 127. A compound of any one of embodiments 1-21 or a pharmaceutically acceptable salt, solvate or prodrug thereof, or a compound of formula VII or a pharmaceutically acceptable salt, solvate or prodrug thereof, for treating an acute inflammatory condition in a subject, said condition being associated with coronavirus infection.

[0558] 128. A compound of any one of embodiments 1-21 or a pharmaceutically acceptable salt, solvate or prodrug thereof, or a compound of formula VII or a pharmaceutically acceptable salt, solvate or prodrug thereof, for treating a subject with CRS or cytokine storm, wherein the CRS or cytokine storm is associated with coronavirus infection.

[0559] 129. A compound of any one of embodiments 1-21 or a pharmaceutically acceptable salt, solvate or prodrug thereof, or a compound of formula VII or a pharmaceutically acceptable salt, solvate or prodrug thereof, for treating SARS in a subject, wherein the SARS is associated with coronavirus infection.

[0560] To enable the present invention to be readily understood and practiced, specific preferred embodiments will now be described through the following non-limiting examples.

[0561] Example

[0562] All starting materials, reagents and equipment used are commercially available (e.g. from Sigma-Aldrich (Burlington, MA, USA) and obtained from commercial sources, unless otherwise indicated.

[0563] Unless otherwise stated, all reactions were carried out in a nitrogen atmosphere. A Bruker Ultraspin 400MHz spectrometer was used. 1 H 400 MHz; 13 NMR spectra were collected at 100 MHz. Rapid chromatography was performed using Merck silica gel 60 μm. The reaction was monitored by thin-layer chromatography (TLC) using Merck silica gel 60 μm TLC plates with an aluminum backing and containing F254 fluorescent indicator. The compounds were developed on the TLC plates with UV light and then stained with potassium permanganate solution (containing KMnO4 (1.5 g), K2CO3 (10 g), and 10% NaOH solution (1.25 mL), diluted with deionized water to 200 mL) or with a solution containing 10% H2SO4 in ethanol. Unless otherwise specified, all organic extracts were dried over anhydrous magnesium sulfate, filtered, and then the solvent was removed under reduced pressure.

[0564] Example 1 - Synthesis of 2,3,3',4',6-penta-O-acetylsucralose (compound 1)

[0565] 1

[0567] Acetic anhydride (15 mL, 100 mmol) was added to a stirred solution of sucralose (6.1 g, 15 mmol) in pyridine (80 mL). The mixture was then stirred at room temperature (rt) for 24 h and concentrated by co-evaporation of pyridine and toluene. The crude material was dissolved in ethyl acetate (EtOAc) (100 mL) and washed with 1 M HCl (2 × 100 mL), 5% NaHCO3 aqueous solution (100 mL), and brine (50 mL), dried (MgSO4), and concentrated under reduced pressure to give a glassy solid. The crude material was recrystallized from toluene to give 2,3,3',4',6-penta-O-acetylsucralose 1 (8.5 g, 97%) as needle-like crystals. 1 H NMR (CDCl3, 400 MHz): 5.67 (d, 1H, J 3’,4’ =6.4 Hz, H-3'), 5.65 (d,J 1,2 =2.0 Hz, H-1), 5.29-5.27 (m, 2H, H-2, H-3), 4.58-4.52 (m, 2H, H-4, H-5), 4.27-4.19 (m, 3H, H-5', H-6a, H-6b), 3.75 (dd, 2H, J 5’,6’ =6.2 Hz, J 6’a,6’b =0.7 Hz, H-6'a, H-6'b), 3.69, 3.57 (ABq, 2H, J =12. 1 Hz, H-1'), 2.12(s, 3H, CH3), 2.11 (s, 3H, CH3), 2.09 (s, 3H, CH3), 2.08 (s, 3H, CH3), 2.07(s, 3H, CH3); 13 C NMR (CDCl3, 100 MHz): 170.4, 170.2, 170.0, 169.8, 169.6 (5× C OCH3), 104.4 (C-2'), 90.7 (C-1), 80.8 (C-5'), 77.2, 76.1 (C-4'), 75.9 (C-3'), 68.0 (C-2), 67.8 (C-4), 66.9 (C-3), 63.6 (C-6), 59.0 (C-5), 44.5 (C-1'), 43.9 (C-6'), 20.8, 20.7 (×2), 20.5 (5×CO C H3).

[0568] Example 2 - Synthesis of 2,3,3',4',6-penta-O-acetyl-6'-azidosucralose (compound 2)

[0569] 2

[0571] 2,3,3',4',6-penta-O-acetylsucralose 1 (prepared according to Example 1) (35 g, 57 mmol) was dissolved in dimethyl sulfoxide (DMSO) (100 mL), followed by the addition of sodium azide (10 g, 154 mmol), and the resulting solution was heated at 80 °C for 48 h. The reaction mixture was cooled to room temperature and partitioned between diethyl ether (Et2O) (300 mL) and water (300 mL). The organic layer was washed twice with water (2 x 400 mL), and all three aqueous layers were re-extracted with Et2O (200 mL). The combined organic layers were dried (MgSO4), filtered, and concentrated under reduced pressure. The resulting yellow gel was dissolved in a minimal amount of EtOAc (approximately 50 mL), followed by the addition of a small amount of petrol until turbidity was observed. Seed crystals were added and scraped, resulting in a precipitate, which continued to form with the slow addition of more petrol and scraping. The resulting precipitate was filtered and washed with 1:4 EtOAc / gasoline to produce 2,3,3',4',6-penta-O-acetyl-6'-azidosucralose 2 (20 g, 56%) as a white powder. 1 H NMR (CDCl3, 400 MHz): 5.68 (d, 1H, J 3’-4’ =6.9 Hz, H-3'), 5.64 (d, 1H, J 1,2 =3.6 Hz, H-1), 5.32 (dd, 1H, J 2,3 =10.7 Hz, H-3), 5.32-5.29 (m, 1H, H-4'), 5.25 (dd, 1H, H-2), 4.58-4.49 (m, 2H, H-4, H-5), 4.24-4.20 (m, 2H, H-6a, H-6b), 4.15-4.08 (m, 1H, H-5'), 3.69, 3.58 (ABq, 2H, J =12.1 Hz, H-1'), 3.64-3.56 (m, 1H, H-6'a), 3.50 (dd, 1H, J 6a’,6b’ =4.1 Hz, J 5,6b’ =13.3 Hz, H-6b'), 2.10(s, 3H, CH3), 2.09 (s, 3H, CH3), 2.08 (s, 3H, CH3), 2.06 (s, 3H, CH3), 2.05(s, 3H, CH3);13 C NMR (CDCl3, 100 MHz): 170.3, 170.1, 170.0, 169.9, 169.7 (5× C OCH3), 104.1 (C-2'), 90.8 (C-1), 80.0 (C-5'), 75.6 (C-3'), 75.1 (C-4'), 67.9(C-3), 67.7 (C-5), 67.0 (C-2), 63.4 (C-6), 58.9 (C-4), 52.5 (C-6'), 44.5 (C-1'), 20.7, 20.6, 20.6, 20.6, 20.4 (5×CO C H3); HRMS (ESI); m / z [M + Na] + Calculate without C 22 H 29 Cl2N3O 13 Na; measured value is 636.0994.

[0572] Example 3 - Synthesis of 1,2,3,4,6-penta-O-acetyl-D-pyranoylmannose (compound 3)

[0573] 3

[0575] Acetic anhydride (20.7 mL, 160 mmol) was added to a mixture of D-mannose (3.0 g, 10.6 mmol) and sodium acetate (1.6 g, 20 mmol). The reaction mixture was then stirred at 80 °C for 4 h and poured into a cold, saturated (saturated) NaHCO3 solution (100 mL). The mixture was then stirred for 1 h, extracted with dichloromethane (DCM) (2 × 100 mL), dried (MgSO4), and concentrated under reduced pressure. 1,2,3,4,6-penta-O-acetyl-D-pyranoylmannose 3 (3.5 g, 85%, α / β 1:0.4) was given as a colorless oil. The crude product was used in subsequent steps without further purification. 1 H NMR (CDCl3,400 MHz): 6.06 (d, 1H, J =1.1 Hz), 5.84 (d, 0.4 H, J =1.3 Hz), 5.46 (dd, 0.4H, J =1.1 Hz, J=3.3 Hz), 5.34-5.32 (m, 2H), 5.28 (d, 0.4H,J =9.9 Hz), 5.25-5.22(m, 1H), 5.11 (dd, 0.4H, J =3.3 Hz, J=10.0 Hz), 4.28 (dd, 0.4H, J =5.4 Hz, J =12.5 Hz), 4.26 (dd, 1H, J=4.9 Hz, J=12.3 Hz), 4.15-3.98 (m, 2.4H), 3.78 (ddd,0.4H, J =2.4 Hz, J=5.3 Hz, J =7.8 Hz), 2.19 (s, 1.2H, OCH3), 2.16 (s, 3H, OCH3), 2.15 (s, 3H, OCH3), 2.08 (s, 1.3H, OCH3), 2.07 (s, 4H, OCH3), 2.03 (s, 4H,OCH3), 1.99 (s, 4H, OCH3).

[0576] Example 4 -Propyneyl 2,3,4,6-tetra- O Synthesis of α-acetyl-A-D-mannopyranoside (compound 4)

[0577] 4

[0579] Propyrrolizidine ether (1.1 mL, 19.2 mmol) was added to a stirred solution of 1,2,3,4,6-penta-O-acetyl-D-pyranoylmannose 3 (prepared according to Example 3) (1.5 g, 3.8 mmol) in a dry DCM (20 mL). The mixture was cooled to 0 °C and BF3.OEt2 (4.9 mL, 38.4 mmol) was added dropwise. The resulting mixture was stirred for 15 min, allowed to reach room temperature, and stirred for 24 h. The reaction mixture was diluted with DCM (100 mL) and poured into an ice-cold saturated NaHCO3 (100 mL) solution. The resulting mixture was stirred until no more bubbles were produced. The organic phase was then separated, washed with (3 × 50 mL) saturated NaHCO3 and water (100 mL), dried (MgSO4), and concentrated under reduced pressure. The crude material was then purified by recrystallization from petroleum spirit (Pet. Sp.) / EtOAc (8:1) to obtain propynyl 2,3,4,6-tetra- as colorless needle-like crystals. O-Acetyl-α-D-mannopyranoside 4 (1.2 g, 85%). 1 H NMR (CDCl3, 400MHz): 5.33 (dd, 1H, J 2,3 =3.2 Hz, J 3,4 =9.8 Hz, H-3), 5.29 (d, 1H, J 3,4 = J4,5 =9.3Hz, H-4), 5.26 (dd, 1H, J 1,2 =1.8 Hz, J 2,3 =3.2 Hz, H-2), 5.01 (d, 1H, J 1,2 =1.8Hz, H-1), 4.30-4.24 (m, 3H, H-6a, C H 2C≡CH), 4.09 (dd, J 5,6b =2.5 Hz, J 6a,6b =12.3Hz, H-6b), 4.00 (ddd, 1H, J 4,5 =9.2 Hz, J 5,6b =2.5 Hz, J 5,6a =5.3 Hz, H-5), 2.45 (t, 1H, J =2.4 Hz, CH2C≡C H ), 2.14 (s, 3H, CH3), 2.09 (s, 3H, CH3), 2.02 (s, 3H, CH3), 1.97 (s, 3H, CH3); 13 C NMR (CDCl3, 100 MHz): 170.6, 169.9, 169.8, 169.7 (4×CH3 C O), 96.3 (C-1), 77.9 (OCH2 C ≡CH), 75.6 (OCH2C≡ C H), 69.3 (C-2), 69.0 (C-3), 68.8 (C-5), 66.1 (C-4), 62.3 (C-6), 55.0 (O CH2C≡CH), 20.9, 20.7 (2×), 20.6 (4×) C H3CO); HRMS (ESI); m / z [M + Na] + Calculated as C 17 H 22 O 10 Na; measured value is 409.1121.

[0580] Example 5 -Propyne-2,3,4-tri- O Synthesis of benzoyl-6-toluenesulfonyl-α-D-mannopyranoside (compound 5)

[0581] 5

[0583] A catalytic amount of sodium metal (approximately 4 mg) was dissolved in methanol (MeOH) (50 mL). The resulting solution was then added to propynyl 2,3,4,6-tetra- O Acetyl-α-D-mannopyranoside 4 (prepared according to Example 4) (50.0 g, 130 mmol) was added to a solution of MeOH (500 mL), and the mixture was stirred at 45 °C for 1 h. The reaction mixture was then subjected to pre-washing with an acidic ion exchange resin (IR-120 H). + Neutralize, filter, and concentrate under reduced pressure. Dissolve the residue in a minimal amount of methanol. Precipitate the product by adding ethyl acetate fractionally. Filter the mixture to produce a pale yellow / brown solid (25.5 g, 90%). Dissolve the crude material in anhydrous pyridine (200 mL) and add toluenesulfonyl chloride (30 g, 157 mmol) to the reaction mixture, allowing it to stand at room temperature for 18 h. Add benzoyl chloride (50 mL, 60 g, 426 mmol) and allow it to stand at room temperature for 4 h. Add water (50 mL) to the reaction mixture and evaporate it to dryness. Dissolve the resulting crude residue in EtOAc (1000 mL) and wash with 1 M HCl (2 × 500 mL), saturated NaHCO3 (500 mL), dry (MgSO4), and concentrate under reduced pressure. The crude material was purified by rapid column chromatography on silica gel (15-35% EtOAc / Pet.Sp.) to provide propynyl 2,3,4-tri- as a colorless oil. O -Benzoyl-6-toluenesulfonyl-α-D-mannopyranoside 5 (7.3 g, 70%). 1 H NMR (CDCl3, 400 MHz): 8.06 (dd, 2H, J =1.1 Hz,J =8.1 Hz,H-OBz), 7.90 (dd, 2H, J =0.9 Hz, J =8.1 Hz, H-OBz), 7.78 (dd, 2H, J =1.2 Hz, J =8.3 Hz, H-OBz), 7.62-7.18 (m, 8H, H-OBz), 7.13 (s, 2H, H-OTPS), 5.87 (dd, 1H, J 2,3 =3.3 Hz, J 3,4 =10.0 Hz, H-3), 5.75 (dd, 1H, J 3,4 = J 4,5 =10.0 Hz, H-4), 5.67 (dd,1H, J 1,2 =1.8 Hz, J 2,3 =3.3 Hz, H-2), 5.26 (d, 1H, J 1,2 =1.8 Hz, H-1), 4.47-4.40(m, 1H, H-5), 4.32 (t, 2H, J =2.9 Hz, C H 2C≡CH), 4.29-4.25 (m, 2H, H-6), 4.08(h, 2H, J =6.9 Hz, J =13.3 Hz, J =20.1 Hz, C H - i -pr), 2.86 (h, 1H, J =6.9 Hz, J =13.3 Hz, J =20.1 Hz, C H - i -pr), 2.48 (t, 1H, J =2.4 Hz, CH2C≡C H ), 1.22 (s, 3H,CH3), 1.19 (s, 3H, CH3), 1.18 (s, 3H, CH3), 1.17 (s, 3H, CH3), 1.15 (s, 3H,CH3); 13C NMR (CDCl3, 100 MHz): 165.5, 165.3, 165.2 (3×C=O), 153.8, 150.9,133.5, 129.9, 129.8, 129.7, 128.6, 128.4, 128.2, 123.7 (Ar-OTPS), 95.9 (C-1),77.9, 77.2 (OCH2 C ≡CH), 75.8 (OCH2C≡ C H), 70.2 (C-2), 69.7 (C-3), 69.6 (C-5), 67.8 (C-6), 67.0 (C-4), 55.1 (O C H2C≡CH), 34.1 ( C H- i -pr), 29.6 ( C H- i -pr), 24.6, 24.5, 23.4 (6× C H3); HRMS (ESI); m / z [M + Na] + Calculated as C 44 H 48 O 11 NaS; measured value is 819.2844.

[0584] Example 6 - Synthesis of Compound 10 (G24)

[0585] 10

[0587] The general synthetic scheme for compound 10 is shown below:

[0588]

[0589]

[0590] Program 1 for synthesizing SO3.Py

[0591] Chlorosulfonic acid (1.6 mL) was added dropwise to a solution of pyridine (4 mL) in DCM (20 mL) at 0 °C, and the mixture was stirred for 30 min. The reaction mixture was then filtered, and the precipitate was washed with ice-cold water (3 × 20 mL), ice-cold saturated NaHCO3 (2 × 10 mL), ice-cold water (2 × 10 mL) (until the pH of the filtrate was 7), cold ethanol (EtOH) (2 × 20 mL), cold toluene (10 mL), and cold DCM (10 mL). The precipitated SO3·Py was then thoroughly dried under high vacuum and used within 24 h.

[0592] General procedure for click response 1

[0593] Sodium ascorbate [2 equivalents] and copper sulfate pentahydrate (1 equivalent) were added to a mixture of an azide analog (1 equivalent) and an alkyne analog in 1,4-dioxane / H₂O (3:1). The reaction mixture was then stirred at room temperature for 18 h and filtered on a diatomaceous earth mat. The filtrate was concentrated under reduced pressure and the crude substance was dissolved in EtOAc. The organic layer was washed with 1M HCl and saturated NaHCO₃, dried (MgSO₄), and concentrated under reduced pressure. The resulting crude substance was purified by rapid column chromatography on silica gel to obtain the desired compound.

[0594] General Procedure 1 for the Hydrolysis of Acetates and Benzoates

[0595] The oligosaccharide was dissolved in methanol, followed by the addition of a 30% NH3 aqueous solution over 10 minutes to ensure that the starting material did not precipitate from the solution. The resulting solution was allowed to stand at room temperature (24–48 h), then concentrated and pre-adsorbed onto silica gel. Rapid chromatography was performed to remove acetamide and benzamide, yielding a polyol as a foam.

[0596] General Procedure for Sulfation and Dialysis 1

[0597] A polyol (100 mg) was dissolved in dimethylformamide (DMF) (10 mL), followed by the addition of freshly prepared SO3·Py (10 equivalents / hydroxyl). The reaction mixture was allowed to stand at room temperature (5 days) and then quenched with 5 M NaOH solution (2 equivalents / SO3·Py). The resulting amber-colored mixture was partitioned between water (20 mL) and CH2Cl2 (50 mL). The aqueous layer was washed again with CH2Cl2 and then concentrated. The resulting residue was dissolved in Milli-Q water (5–10 mL) and dialyzed using a cellulose membrane tube (approximately 30 cm) with a molecular weight cutoff (MWCO) of 0.5 kD. The dialysis tube was washed with Milli-Q water before use and the ends were clamped. The tube was placed in 4 L of 0.1 M NaCl solution for 2 h and then in 4 L of Milli-Q water overnight. The dialysis tube expanded significantly, holding approximately 150 mL of overnight dialysate. The contents of the dialysis tube were removed, concentrated to approximately 5 mL, and transferred to a Microsep Advanced tube equipped with a 1 kD MWCO Omega membrane (Pall Corporation, New York, USA). The tube was centrifuged at 5000–7000 g to concentrate to approximately 200 μL, and then washed twice with 3 mL of Milli-Q water. The retained material was then passed through a C18 Sep-Pak elution system, first with water (3 mL) followed by methanol (3 mL) to remove most of the remaining color. The target product was determined by thin-layer chromatography (TLC) (see below) to be present in the aqueous phase, and then lyophilized to obtain glycanic sulfate as a powder.

[0598] Synthesis of Compound 6

[0599] 6

[0601] 2,3,3',4',6-penta-O-acetyl-6'-azidosucralose 2 (prepared according to Example 2) (8.0 g, 13.0 mmol) and propynyl 2,3,4-tri- OThe click reaction of benzoyl-6-toluenesulfonyl-α-D-mannopyranoside 5 (prepared according to Example 5) (8.0 g, 11.7 mmol) (72 h.) yielded a yellow foam, which was dissolved in DMF (100 mL) followed by the addition of sodium azide (1.5 g, 23.0 mmol). The resulting mixture was heated at 60 °C for 48 h and then partitioned between EtOAc (300 mL) and water (300 mL). The aqueous phase was redissolved with EtOAc (200 mL), and the combined organic layers were washed with water (2 x 300 mL), dried (MgSO4), concentrated, and subjected to rapid chromatography (40% - 60% EtOAc / Pet. Sp.) to give azide 6 (12.0 g, 88%) as a colorless foam. 1 H NMR (CDCl3, 400 MHz): δ =8.11-8.07 (m, 2H), 7.97-7.93 (m, 2H), 7.85 (s, 1H), 7.82-7.78 (m, 2H), 7.64-7.57 (m, 1H), 7.55-7.46 (m, 3H), 7.44-7.34 (m, 3H),7.28-7.21 (m, 2H), 5.93-5.84 (m, 2H), 5.75 (d, 1H, J =7.4 Hz), 5.74-5.72 (m,1H), 5.69-5.67 (m, 1H), 4.43 (t, 1H, J =7.3 Hz), 5.37-5.32 (m, 2H), 5.25 (d,1H, J =1.7 Hz), 5.02 (d, 1H, J =12.5 Hz), 4.89 (dd, 1H, J =3.7 and 14.4 Hz), 4.87 (d, 1H, J =12.5 Hz), 4.76 (dd, 1H, J =9.1 and 14.4 Hz), 4.64-4.58 (m, 2H), 4.49 (ddd, 1H, J =3.6, 7.2 and 8.9 Hz), 4.41-4.32 (m, 1H), 4.36 (dd, 1H, J =4.3 and 11.8 Hz), 4.27 (dd, 1H, J=6.9 and 11.8 Hz), 3.64 (d, 1H, 11.9 Hz), 3.55 (d, 1H, 11.9Hz), 3.53 (m, 2H), 2.16 (s, 3H), 2.12 (s, 3H), 2.11 (s, 3H), 2.09 (s, 3H),2.01 (s, 3H).

[0602] Synthesis of Compound 7

[0603] 7

[0605] Azide 6 (12.0 g, 10.3 mmol) and propynyl 2,3,4-tri- O The click reaction (24 h.) of benzoyl-6-toluenesulfonyl-α-D-mannopyranoside 5 (prepared according to Example 5) (7.0 g, 10.2 mmol) yielded a foam, which was dissolved in DMF (100 mL) followed by the addition of sodium azide (1.5 g, 23.0 mmol). The resulting mixture was heated at 60 °C for 24 h and then partitioned between EtOAc (300 mL) and water (300 mL). The aqueous phase was redissolved with EtOAc (200 mL), and the combined organic layers were washed with water (2 x 300 mL), dried (MgSO4), concentrated, and subjected to rapid chromatography (50% - 80% EtOAc / Pet. Sp.) to give azide 7 (13.5 g, 77%) as a pale yellow foam. 1 H NMR (CDCl3, 400 MHz): δ =8.09-7.98 (m, 7H), 7.95-7.89 (m,2H), 7.82-7.74 (m, 5H), 7.67-7.57 (m, 2H), 7.56-7.45 (m, 6H), 7.44-7.32 (m,6H), 7.26-7.20 (m, 2H), 5.93-5.73 (m, 6H), 5.66 (dd, 1H, J =1.6 and 3.3 Hz), 5.61(dd, 1H, J =1.8 and 3.1 Hz), 5.42 (t, 1H, J =7.3 Hz), 5.39 (dd, 1H, J =3.5 and 10.7 Hz), 5.33 (dd, 1H, J=3.3 and 10.8 Hz), 5.24–5.19 (m, 1H), 4.97 (d, 1H, J =12.4Hz), 4.88-4.56 (m, 10H), 4.47 (ddd, 1H, J =3.6, 7.2 and 8.9 Hz), 4.37-4.30 (m, 1H), 4.33 (dd, 1H, J =4.5 and 11.6 Hz), 4.26 (dd, 1H, J =7.3 and 11.8 Hz), 3.65, 3.61 (ABq, 2H, J =12.2 Hz), 3.65-3.44 (m, 2H), 2.16 (s, 3H), 2.09 (s, 3H), 2.09 (s, 3H), 2.07 (s, 3H), 1.96 (s, 3H).

[0606] Synthesis of Compound 8

[0607] 8

[0609] A click reaction (48 h.) of azide 7 (5.0 g, 2.9 mmol) and 1,5-hexanediyne (50% in pentane, 280 μL, 1.4 mmol) was carried out using General Procedure 1, yielding an incomplete reaction indicated by TLC. A second portion of copper sulfate pentahydrate and sodium ascorbate were added, and the reactants were heated at 50 °C for 4 h. The reaction was then indicated by TLC as complete. Following standard post-processing according to General Procedure 1, rapid chromatography (80%–100% EtOAc / Pet. Sp.) was performed to give dimer 8 (3.0 g, 59%) as a light brown foam. 1 H NMR (CDCl3, 400 MHz): δ =8.05-7.95 (m,5H), 7.95-7.85 (m, 4H), 7.82-7.76 (m, 2H), 7.75-7.70 (m, 3H), 7.67-7.30 (m,15H), 7.25-7.18 (m, 4H), 5.93-5.82 (m, 2H), 5.81-5.68 (m, 3H), 5.67-5.58 (m,2H), 5.54 (dd, 1H, J =1.6 and 3.1 Hz), 4.45-4.36 (m, 2H), 5.32 (dd, 1H, J=3.3 and 10.9 Hz), 5.21 (d, J =1.3 Hz), 5.15 (d, J =1.3 Hz), 4.88-4.51 (m, 13H), 4.50-4.42 (m, 1H), 4.33 (dd, 1H, J =4.6 and 11.9 Hz), 4.24 (dd, 1H, J =7.0 and 11.9 Hz), 3.68 (d, 1H, J =12.0 Hz), 3.61 (d, 1H, J =12.0 Hz), 3.09-3.01 (m, 2H), 2.15 (s,3H), 2.07 (s, 3H), 2.07 (s, 3H), 2.05 (s, 3H), 1.95 (s, 3H).

[0610] Synthesis of Compound 9

[0611] 9

[0613] According to general procedure 1, the acetate hydrolysis of compound 8 (2.8 g, 0.8 mmol) was carried out using methanol (200 mL) and 30% NH3 aqueous solution (30 mL) for 24 h, and oligosaccharide 9 (1.2 g, 81%) as a light brown foam was produced after rapid chromatography (50% - 100% MeOH / EtOAc). 1 H NMR (D2O, 400 MHz): δ =7.89 (bs, 2H),7.79 (s, 1H), 5.46 (d, 1H, J =4.1 Hz), 4.94-4.85 (m, 2H), 4.62-4.51 (m, 2H), 4.48-4.32 (m, 4H), 4.31-4.13 (m, 6H), 3.95 (dd, 1H, J =10.3 and 4.0 Hz), 3.92 (dd, 1H, J =3.4 and 1.6 Hz), 3.90-3.74 (m, 6H), 3.70 (s, 2H), 3.65 (dd, 1H), J =9.5 and 3.4 Hz), 3.61-3.53 (m, 2H), 2.97 (s, 2H).

[0614] Synthesis of Compound 10

[0615] 10

[0617] Sulfated deprotected octamer 9 (100 mg, 0.05 mmol) was dialyzed according to general procedure 1, followed by dialysis, to produce octamer 10 (20 mg, 9%) as a colorless powder. 1 H NMR (D2O, 400 MHz): δ =8.15 (s,1H), 8.07 (s, 1H), 7.93 (bs, 1H), 5.88 (d, J =3.6 Hz, 1H), 5.39 (d, J =7.9 Hz, 1H), 5.30 (d, 1H, J =1.9 Hz), 5.24 (d, 1H, J =1.8 Hz), 5.17-5.05 (m, 2H), 5.05-4.85 (m, 8H), 4.78-4.61 (m, 4H), 4.59-4.21 (m, 11H), 4.00, 3.93 (ABq, 2H, J =12.5 Hz).

[0618] Example 7 - Synthesis of propargyl maltotriose (compound 11)

[0619] 11

[0621] The general synthesis scheme is shown below:

[0622]

[0623] Boron trifluoride ether (46 mL) was added dropwise to peracetic acid maltotriose at 0 °C (according to Su et al. (2019)). Eur J Med Chem The mixture of propargyl maltotriose 11 (13.3 g, 13.8 mmol) and propargyl alcohol (8.5 mL, 137 mmol) in a dry DCM (500 mL) was prepared according to the procedure of 182:111622. The mixture was then stirred at room temperature for 48 h. Saturated NaHCO3 (10 mL) was then added to the reaction mixture and stirred for another 10 min. The organic layer was extracted, dried (MgSO4), and concentrated under reduced pressure. The resulting crude product was purified by rapid column chromatography (3:1 EtOAc: Pet. Sp.) to give propargyl maltotriose 11 (6.0 g, 45%) as a glassy solid. 1H NMR (CDCl3, 400 MHz): 5.43-5.22(m, 6H), 5.04 (t, 1H, J =10.0 Hz), 4.86-4.76 (m, 3H), 4.71 (dd, 1H, J =4.1 Hz,J=10. 4 Hz), 4.51-4.39 (m, 2H), 4.33 (d, 2H, J =2.3 Hz), 4.32-4.13 (m, 4H), 4.06-3.87 (m, 6H), 3.77-3.69 (m, 1H), 2.44 (t, 1H, J =2.4 Hz), 2.15, 2.13,2.07, 2.02, 2.02, 2.00, 1.98, 1.97 (×2) (s, 3H, 9×CH3); 13 C NMR (CDCl3, 100MHz): 170.6 (×2), 170.5 (×2), 170.3, 170.1, 169.8, 169.7 (×2), 169.4,97.5, 95.7 (×2), 77.2, 75.5, 75.2, 73.7, 72.5, 72.2, 71.8, 71.7, 70.4, 70.1,69.4, 68.9, 68.5, 67.9, 62.8, 62.3, 61.4, 60.4, 55.8, 20.9, 20.8, 20.6, 20.5,14.2; HRMS (ESI); m / z [M + Na] + Calculated as C 41 H 54 O 26 Na; measured value is 385.2792.

[0624] Example 8 -Propyneyl-2,3,4-tri- O Synthesis of benzoyl-6-2,4,6-triisopropylbenzenesulfonyl-α-D-mannopyranoside (compound 12)

[0625] 12

[0627] propynyl 2,3,4,6-tetra- OAcetyl-α-D-mannopyranoside 4 (prepared according to Example 4) (5.0 g, 13 mmol) was added to a solution of sodium metal (approximately 4 mg) in MeOH (50 mL), and the mixture was stirred at room temperature for 2 h. The reaction mixture was then subjected to a pre-washed acidic ion exchange resin (IR-120 H). + The mixture was neutralized, filtered, and concentrated under reduced pressure. The crude material was dissolved in anhydrous pyridine (50 mL) and cooled to 0 °C. 2,4,6-Triisopropylbenzenesulfonyl chloride (4.7 g, 16 mmol) was added to the reaction mixture and allowed to warm to room temperature, then stirred for 18 h. The reaction mixture was then cooled to 0 °C and benzoyl chloride (5.0 mL, 47 mmol) was added and stirred at room temperature for 24 h. The reaction mixture was concentrated under reduced pressure by co-evaporation of pyridine and toluene. The resulting crude residue was dissolved in EtOAc (100 mL), washed with 1 M HCl (2 × 50 mL) and saturated NaHCO3 (50 mL), dried (MgSO4), and concentrated under reduced pressure. The crude material was purified by rapid column chromatography on silica gel (Pet. Sp. / EtOAc 3:1) to provide propynyl 2,3,4-triisopropylbenzenesulfonyl chloride as a colorless oil. O -benzoyl-6-2,4,6-triisopropylbenzenesulfonyl-α-D-mannopyranoside 12 (7.3 g, 70%). 1 H NMR (CDCl3, 400 MHz): 8.06 (dd, 2H, J =1.1 Hz, J =8.1 Hz, H-OBz), 7.90 (dd, 2H, J =0.9 Hz, J =8.1 Hz, H-OBz), 7.78 (dd, 2H, J =1.2 Hz, J =8.3 Hz, H-OBz), 7.62-7.18(m, 8H, H-OBz), 7.13 (s, 2H, H-OTPS), 5.87 (dd, 1H, J 2,3 =3.3 Hz, J 3,4 =10.0 Hz, H-3), 5.75 (dd, 1H, J 3,4 = J 4,5 =10.0 Hz, H-4), 5.67 (dd, 1H, J 1,2 =1.8 Hz,J 2,3 =3.3Hz, H-2), 5.26 (d, 1H, J 1,2 =1.8 Hz, H-1), 4.47-4.40 (m, 1H, H-5), 4.32 (t, 2H, J =2.9 Hz, C H 2C≡CH), 4.29-4.25 (m, 2H, H-6), 4.08 (h, 2H, J =6.9 Hz, J =13.3Hz, J =20.1 Hz, C H - i -pr), 2.86 (h, 1H, J =6.9 Hz, J =13.3 Hz, J =20.1 Hz, C H - i -pr), 2.48 (t, 1H, J =2.4 Hz, CH2C≡C H ), 1.22 (s, 3H, CH3), 1.19 (s, 3H, CH3),1.18 (s, 3H, CH3), 1.17 (s, 3H, CH3), 1.15 (s, 3H, CH3); 13 C NMR (CDCl3, 100MHz): 165.5, 165.3, 165.2 (3×C=O), 153. 8, 150.9, 133.5, 129.9, 129.8,129.7, 128.6, 128.4, 128.2, 123.7 (Ar-OTPS), 95.9 (C-1), 77.9, 77.2 (OCH2 C ≡CH), 75.8 (OCH2C≡ C H), 70.2 (C-2), 69.7 (C-3), 69.6 (C-5), 67.8 (C-6), 67.0(C-4), 55.1 (O C H2C≡CH), 34.1 ( C H- i -pr), 29.6 ( C H- i -pr), 24.6, 24.5, 23.4 (6× CH3); HRMS (ESI); m / z [M + Na] + Calculated as C 44 H 48 O 11 NaS; measured value is 819.2844.

[0628] Example 9 - Synthesis of Compound 20 (G22)

[0629] 20

[0631] The general synthetic scheme for compound 20 is shown below.

[0632]

[0633]

[0634] Program 2 for synthesizing SO3.Py

[0635] Chlorosulfonic acid (1.6 mL) was added dropwise to a solution of pyridine (4 mL) in DCM (20 mL) at 0 °C, and the mixture was stirred for 30 min. The reaction mixture was then filtered, and the precipitate was washed with ice-cold water (3 × 20 mL), ice-cold saturated NaHCO3 (2 × 10 mL), ice-cold water (2 × 10 mL) (until the pH of the filtrate was 7), cold EtOH (2 × 20 mL), cold toluene (10 mL), and cold DCM (10 mL). The precipitated SO3·Py was then thoroughly dried under high vacuum and used within 24 h.

[0636] General Program 2 for Click Response

[0637] Sodium ascorbate (2 equivalents) and copper sulfate pentahydrate (1 equivalent) were added to a mixture of an azide analog (1 equivalent) and an alkyne analog (1 equivalent / N3) in 1,4-dioxane / H2O (3:1). The reaction mixture was then stirred at room temperature for 18 h and filtered on a diatomaceous earth mat. The filtrate was concentrated under reduced pressure, and the crude substance was dissolved in EtOAc. The organic layer was washed with 1M HCl and saturated NaHCO3, dried (MgSO4), and concentrated under reduced pressure. The resulting crude substance was purified by rapid column chromatography on silica gel to obtain the desired compound.

[0638] General Procedure 2 for the Hydrolysis of Acetates and Benzoyl Esters

[0639] A solution of 30% NH3 aqueous solution (3-5 mL) and oligosaccharide in methanol (20 mL) was stirred at room temperature for 48 h. The reaction mixture was concentrated under reduced pressure, and the crude material was dissolved in ultrapure water (5-10 mL) and washed with EtOAc (5 × 10 mL). The aqueous layer was concentrated to obtain the pure compound.

[0640] General procedure 2 for sulfation

[0641] Freshly prepared SO3·Py (10 equivalents / OH) complex was added to a solution of starting material in dry DMF (10 mL) maintained under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 2 days and adjusted to alkalinity with 5 M NaOH (2 equivalents / SO3·Py). The mixture was then concentrated under reduced pressure, and the crude material was dissolved in ultrapure water (10–20 mL) and dialyzed according to a standard procedure.

[0642] General Procedure for Dialysis 2

[0643] Commercially available cellulose membrane dialysis tubing with MWCO at 0.5 kD, 1 kD, and 2 kD was used for dialysis. Before use, the tubing was washed with milliQ water (for approximately 5–10 minutes). One end of the tubing was tied, and the sample was loaded into the tubing. The tubing was then clamped shut, leaving almost no space for air bubbles. A 3 L flask equipped with a stir bar was filled with milliQ water and placed on a magnetic stir plate at room temperature. The stirring was set so that the dialysis bag floated slowly on top of the solution in the flask. After 2 hours, the water was replaced with fresh milliQ water, and dialysis was continued for 2 days, with the dialysis water replaced every 18 hours. The dialysis bag was then removed from the flask, and its contents were collected and freeze-dried.

[0644] 2,3,4,6-Titanium O Synthesis of α-acetyl-D-mannopyranoside (compound 13)

[0645] 13

[0647] Octanol (4.0 mL, 25.5 mmol) was added to a stirred solution of 1,2,3,4,6-penta-O-acetyl-D-pyranoylmannose 3 (prepared according to Example 3) (5.6 g, 14.3 mmol) in dry DCM (100 mL). The mixture was cooled to 0 °C and boron trifluoride diethyl ether (BF3.OEt2) (16.0 mL, 112.0 mmol) was added dropwise. The resulting mixture was stirred for 15 min, brought to room temperature, and stirred for 24 h. The reaction mixture was diluted with DCM (100 mL) and poured into an ice-cold saturated NaHCO3 (100 mL) solution. The resulting mixture was stirred until no more bubbles were produced. The organic phase was then separated and washed with (3 × 50 mL) saturated NaHCO3 and water (100 mL), dried (MgSO4), and concentrated under reduced pressure. The crude material was then dissolved in MeOH (50 mL) and a catalytic amount of sodium was added to the mixture. The reactants were stirred at room temperature for 2 hours and then rinsed with a pre-washed acidic ion exchange resin (IR-120 H). + The mixture was neutralized, filtered, and concentrated under reduced pressure. The crude material was purified by rapid column chromatography (EtOAc / MeOH 3:1) to give 13 (2.5 g, 60%) as a colorless oil.

[0648] 2,3,4-Synki O Synthesis of α-acetyl-6-2,4,6-triisopropylbenzenesulfonyl-α-D-mannopyranoside (compound 14)

[0649]

[0650] The crude material from 13 above (880 mg, 3.00 mmol) was dissolved in anhydrous pyridine (50 mL) and cooled to 0 °C. 2,4,6-Triisopropylbenzenesulfonyl chloride (1.18 g, 3.9 mmol) was added to the reaction mixture and allowed to warm to room temperature, then stirred for 18 h. The reaction mixture was then cooled to 0 °C and acetic anhydride (2 mL) was added, and the mixture was stirred at room temperature for 24 h. The reaction mixture was concentrated under reduced pressure by co-evaporation of pyridine and toluene. The resulting crude residue was dissolved in EtOAc (100 mL), washed with 1 M HCl (2 × 50 mL) and saturated NaHCO3 (50 mL), dried (MgSO4), and concentrated under reduced pressure. The crude material was purified by rapid column chromatography on silica gel (Pet. Sp. / EtOAc 3:1) to provide pyranomannoside 14 (1.7 g, 89%) as a colorless oil. 1H NMR (CDCl3, 400 MHz): 7.18 (s, 2H, Ar-OTPS), 5.33 (dd, 1H, J 2,3 =3.5 Hz, J 3,4 =10.0 Hz, H-3), 5.19 (dd, 1H, J 1,2 =1.7 Hz, J 2,3 =3.5 Hz, H-2), 5.11 (t, 1H, J 3,4 = J 4,5 =10.0 Hz, H-4), 4.72 (d, 1H, J 1,2 =1.7Hz, H-1), 4.17-4.05 (m, 5H, H-5), 3.69-3.62 (m, 1H, H-6a), 3.41-3.33 (m, 1H,H-6b), 2.91 (h, 1H, J =6.9 Hz, J =13.7 Hz, J =20.1 Hz, C H - i -pr), 2.09 (s, 3H,CH3), 1.98 (s, 3H, CH3), 1.97 (s, 3H, CH3), 1.67-0.69 (m, 36H, 7×CH2, CH3,OCH3); 13 C NMR (CDCl3, 100 MHz): 170.0, 169.8 (×2), 153.8, 150.9, 129.2,123.7, 97.2 (C-1), 69.7 (C-2), 67.0 (C-3), 68.6 (C-5), 68.5 (C-6), 67.7, 66.7(C-4), 34.2, 31.8, 29.7, 29.6, 29.3, 29.2, 29.1, 26.0, 24.7, 24.6, 23.5,22.6, 20.8, 20.6, 20.5, 14.1.

[0651] Synthesis of octyl 2,3,4-tri-O-acetyl-6-azido-α-D-mannopyranoside (compound 15)

[0652] 15

[0654] To a solution of 14 (2.5 g, 3.7 mmol) in dry DMF (20 mL), NaN3 (800 mg, 20 mmol) was added, and the mixture was stirred at 60 °C for 24 h. The reaction mixture was then decanted, and the residue remaining in the flask was washed with EtOAc (3 × 5 mL). The combined organic layers were then concentrated under reduced pressure. The crude substance was dissolved in EtOAc (100 mL), washed with 1 M HCl (2 × 50 mL) and saturated NaHCO3 (50 mL), dried (MgSO4), and concentrated under reduced pressure. The crude substance was purified by rapid column chromatography (Pet. Sp. / EtOAc 1:1) to give azide 15 as a colorless oil (quantitative). 1 H NMR (CDCl3, 400 MHz): 5.32 (dd, 1H, J=3.5 Hz, J =10.0 Hz, H-3), 5.23-5.15 (m, 2H, H-2, H-4), 4.79 (br-s, 1H, H-1), 3.96-3.88 (m, 1H, J =2.5 Hz, J =6.9 Hz, J=9.6 Hz, H-5), 3.73-3.65 (m, 1H, J =6.6 Hz, J =9.4 Hz), 3.48-3.40 (m, 1H, J =6.5 Hz, J =9.4 Hz), 3.34 (dd, 1H, J =6.7 Hz, J =13.2 Hz, H-6a), 3.24 (dd, 1H, J =2.3 Hz, J =13.1 Hz, H-6b), 2.13 (s, 3H, CH3), 2.02 (s,3H, CH3), 1.96 (s, 3H, CH3), 1.67-0.71 (m, 18H, 7×CH2, CH3); 13C NMR (CDCl3,100 MHz): 171.0, 170.0, 169.8, 97.3, 69.9, 69.6, 68.9, 68.6, 67.3, 60.3,51.1, 31.7, 29.2, 29.1, 26.0, 22.6, 20.9, 20.8, 20.6, 14.1; HRMS (ESI); m / z [M+ Na] + Calculated as C 20 H 33 O8N3Na; measured value is 466.2168.

[0655] Synthesis of Compound 16

[0656] 16

[0658] A click reaction of 12 (prepared according to Example 8) (2.5 g, 3.10 mmol) and mannose azide 15 (2.0 g, 3.10 mmol) was carried out using general procedure 2 to obtain a disaccharide as a glassy solid. Sodium azide (500 mg, 7.40 mmol) was added to a solution of the disaccharide in dry DMF (20 mL), and the mixture was stirred at 60 °C for 18 h under a nitrogen atmosphere. The reaction mixture was then concentrated under reduced pressure, and the resulting crude product was suspended in EtOAc (75 mL). The organic layer was washed with HCl (2 × 75 mL) and saturated NaHCO3, dried (MgSO4), and concentrated. The crude product was then purified by rapid column chromatography to obtain 16 (700 mg, 19%). 1 H NMR (CDCl3, 400 MHz): 8.12-8.06 (m,2H, Ar-H), 7.98-7.91 (m, 2H, Ar-H), 7.85 (s, 1H, H-9), 7.82-7.77 (m, 2H, Ar-H), 7.66-7.59 (m, 1H, Ar-H), 7.55-7.47 (m, 3H, Ar-H), 7.46-7.35 (m, 3H, Ar-H), 7.29-7.22 (m, 2H, Ar-H), 5.93-5.86 (m, 1H, H-4), 5.83 (dd, 1H, J 2,3 =3.2Hz, J 3,4 =10.0 Hz, H-3), 5.67 (dd, 1H, J1,2 =1.7 Hz, J 2,3 =3.2 Hz, H-2), 5.37 (dd,1H, J 2’,3’ =3.5 Hz, J 3’,4’ =10.0 Hz, H-3’), 5.24 (dd, 1H, J 1’,2’ =1.8 Hz, J 2’,3’ =3.4Hz, H-2’), 5.21 (d, 1H, J 1’,2’ =1.7 Hz, H-1’), 5.17 (t, 1H, J 3’,4’ = J 4’,5’ =10.0 Hz,H-4’), 5.04, 4.82 (ABq, 2H, J =12.2 Hz, H-7), 4.77 (d, 1H, J1,2 =1.6 Hz, H-1),4.67 (dd, 1H, J 5’,6’a =2.4 Hz, J 6’a,6’b =14.3 Hz, H-6’a), 4.43 (dd, 1H, J 5’,6’b =9.0Hz, J 6’a, 6’b =14.3 Hz, H-6’b), 4.37-4.31 (m, 1H, H-5), 4.19 (ddd, 1H, J 5’,6’a =2.3Hz, J 6’a, 6’b =9.2 Hz, J =12.0 Hz, H-5’), 3.52 (d, 2H, J =4.4 Hz, H-6), 3.23 (dd,2H, J =1.5 Hz, J =6.5 Hz, J =8.1 Hz, H-7’), 2.17 (s, 3H, OCH3), 2.13 (s, 3H,OCH3), 1.99 (s, 3H, OC H2(CH2)6CH3), 1.48 - 1.37 (m, 2H, OCH2(C H 2)6CH3), 1.27 - 1.14(m, 8H, OCH2(C H 2)6CH3), 1.13 - 1.07 (m, 2H, OCH2(C H 2)6CH3), 0.86 (t, 3H, J =6.8 Hz,OCH2(CH2)6C H 3); 13 C NMR (CDCl3, 100 MHz): 171.1, 170.2, 169.9, 169.7, 165.5,165.4 (×2), 143.4 (C - 8), 133.6 (×2), 133.2, 129.8 (×2), 129.7, 129.1,128.9, 128.7, 128.6, 128.5, 128.3, 124.7 (C - 9), 97.4 (C - 1’), 96.9 (C - 1), 70.6(C - 5), 70.2 (C - 2), 69.6 (C - 3), 69.6 (C - 2’), 69.1 (C - 5’), 68.8 (C - 3’), 68.5(O C H2(CH2)6CH3), 67.7 (C - 4), 67.5 (C - 4’), 61.3 (C - 7), 51.2 (C - 6), 51.1 (C - 6’),31.7 (OCH2( C H2)6CH3), 29.2 (OCH2( C H2)6CH3), 29.1 (×2) (OCH2( C H2)6CH3), 26.0(OCH2( C H2)6CH3), 21.0 (OCH2( C H2)6CH3), 20.8 (×2) (2×OCH3), 20.6 (OCH3), 14.2(OCH2( C H2)6CH3), 14.0 (OCH2(CH2)6 C H3).

[0659] Synthesis of Compound 17

[0660] 17

[0662] A click reaction of disaccharide 16 (700 mg, 0.60 mmol) and 5-hexyn-1-ol (0.09 mL, 0.90 mmol) was performed using general procedure 2 to obtain a disaccharide as a glassy solid. Triethylamine (0.6 mL, 4.5 mmol) and methanesulfonyl chloride (0.07 mL, 0.90 mmol) were added to a solution of the crude material in anhydrous DCM (20 mL), and the mixture was stirred at room temperature for 18 h. The reaction mixture was washed with 1 M HCl (2 × 30 mL) and saturated NaHCO3 (30 mL), dried to MgSO4, and concentrated under reduced pressure. NaN3 (100 mg, 1.7 mmol) was added to a solution of the crude material in anhydrous DMF (20 mL), and the mixture was stirred at 60 °C for 18 h. The reaction mixture was concentrated under reduced pressure, and the crude substance was dissolved in EtOAc (100 mL), washed with 1 M HCl (50 mL) and saturated NaHCO3 (50 mL), dried (MgSO4), and concentrated under reduced pressure. The crude substance was purified by rapid column chromatography (1.5:1 EtOAc / Pet. Sp. to EtOAc) to give azide 17 (370 mg, 58%). 1 H NMR (CDCl3, 400 MHz): 8.04-7.92 (m, 4H, Ar-H), 7.79-7.74 (m, 3H, Ar-H, H-9), 7.66-7.56 (m, 2H, Ar-H, H-10), 7.55-7.44 (m, 3H, Ar-H), 7.44-7.34 (m, 3H, Ar-H), 7.26-7.19 (m, 2H, Ar-H), 5.82 (dd, 1H, J 2’,3’ =3.4 Hz, J 3’,4’ =10.0 Hz, H-3'), 5.73 (t, 1H, J 3’,4’ = J 4’,5’ =9.7 Hz, H-4'), 5.59 (dd, 1H, J 2’,3’ =1.7 Hz, J 1’,2’ =3.3 Hz, H-2'), 5.35 (dd, 1H, J 2,3 =3.5 Hz, J 3,4=10.0 Hz, H-3), 5.23 (dd, 1H, J 1,2 =1.7 Hz, J 2,3 =3.5 Hz, H-2), 5.15 (d, 1H, J 1’,2’ =1.7 Hz, H-1'), 5.14 (t, 1H, J 3,4 = J 4,5 =10.0 Hz, H-4), 4.77 (d, 1H, J 1,2 =1.6 Hz), 4.76-4.67 (m, 1H, H-6'a), 4.66-4.55 (m, 3H, H-6'b, H-6a, H-5'), 4.60, 4.51 (ABq, 2H, H-7), 4.42 (dd, 1H, J 5,6b =8.2 Hz, J 6a,6b =14.4 Hz, H-6b), 4.22-4.15 (m, 1H, H-5), 3.27-3.19 (m, 4H,OC H 2(CH2)6CH3, triazole-(CH2)3C H 2N3), 2.78-2.65 (m, 2H, triazole-(C H 2)3CH2N3), 2.15 (s, 3H,OCH3), 2.11 (s, 3H, OCH3), 1.98 (s, 3H, OCH3), 1.76-1.65 (m, 2H, triazole-(C H 2)3CH2N3), 1.65-1.54 (m, 2H, triazole-(C H 2)3CH2N3), 1.46-1.34 (m, 2H, OCH2( C H2)6CH3),1.21-1.10 (m, 8H, OCH2( C H2)6CH3), 0.83 (t, 3H, OCH2(CH2)6 C H3); 13C NMR (CDCl3, 100MHz): 170.0, 169.9, 169.7, 165.7, 165.2 (×2), 147.5 (C-9), 143.0 (C-11),133.7, 133.6, 133.2, 129.8 (×2), 129.6, 129.0, 128.7, 128.6, 128.5, 128.2,124.7 (C-9), 122.5 (C-10), 97.3 (C-1), 96.5 (C-1'), 70.2 (C-2'), 69.6 (C-5'), 69.5 (C-3', C-2), 68.9 (C-5), 68.8 (C-3), 68.4 (O C H2(CH2)6CH3), 67.9 (C-4'), 67.6 (C-4), 60.7 (C-7), 51.0 (C-6, triazole-(CH2)3 C H2N3), 50.9 (C-6'), 31.7 (OCH2( C H2)6CH3), 29.2 (OCH2( C H2)6CH3), 29.1 (OCH2( C H2)6CH3), 28.2 (triazole-( C H2)3CH2N3),26.4 (triazole-( C H2)3CH2N3), 26.0 (OCH2( C H2)6CH3), 25.0 (triazole-( C H2)3CH2N3), 22.5,20.8 (O C H3), 20.7 (O C H3), 20.6 (O C H3), 14.0 (OCH2(CH2)6 C H3).

[0663] Synthesis of Compound 18

[0664] 18

[0666] A click reaction of azide 17 (130 mg, 0.12 mmol) and propargyl maltotriose 11 (prepared according to Example 7) (120 mg, 0.10 mmol) was carried out using general procedure 2 to obtain oligosaccharide 18 (120 mg, 53%) as a glassy solid. 1H NMR (CDCl3, 400 MHz): δ =8.04-7.91 (m, 4H), 7.81-7.74 (m, 3H),7.68-7.59 (m, 2H), 7.57-7.46 (m, 4H), 7.46-7.35 (m, 3H), 7.26-7.20 (m, 2H),5.83 (dd, 1H, J =3.3 Hz, J =10.0 Hz), 5.72 (m, 1H, J =9.8 Hz), 5.60 (dd, 1H, J =1.7 Hz, J =3.3 Hz), 5.43-5.30 (m, 4H), 5.29-5.20 (m, 3H), 5.17-5.02 (m, 3H),4.95-4.39 (m, 17H), 4.39-4.11 (m, 6H), 4.05 (dd, 1H, J =2.2 Hz, J =12.4 Hz),4.01-3.87 (m, 4H), 3.79-3.70 (m, 1H), 3.24 (t, 2H, J =6.4 Hz), 2.81-2.71 (m,2H), 2.19-1.92 (10×s, 30H), 1.76-1.64 (m, 2H), 1.47-1.36 (m, 2H), 1.30-1.11(m, 10H), 0.84 (t, 3H, J =6.8 Hz); 13C NMR (CDCl3, 100 MHz): δ =170.6 (×2),170.5, 170.4, 170.3, 170.1, 170.0, 169.9, 169.8, 169.7 (×2), 169.6, 169.4,165.7, 165.3, 165.2, 146.9, 143.8, 143.0, 133.8, 133.7, 133.3, 129.9, 129.8,129.7, 129.0, 128.8, 128.7, 128.5 (×2), 128.3, 124.8, 123.0, 122.9, 99.5, 97.3, 96.5, 95.7, 95.6, 77.2, 75.2, 73.7, 72.5, 72.3, 72.0, 71.7, 70.4, 70.2, 70.0, 69.6, 69.5, 69.4, 69.0, 68.9, 68.8, 68.5, 67.9, 67.6, 62.9, 62.8, 62.3, 61.4, 60.8, 51.1 (×2), 50.0, 31.8, 29.5, 29.2, 29.1, 26.1, 26.0, 24.6, 22.6, 20.9, 20.8 (×2), 20.6, 20.5 (×2), 14.1.

[0667] Synthesis of Compound 19

[0668] 19

[0670] Compound 18 (120 mg, 0.05 mmol) was hydrolyzed with its acetate and benzoyl esters according to general procedure 2 to yield oligosaccharide 19 (47 mg, 73%). 1 H NMR (D2O, 400 MHz): δ =7.93 (s, 1H), 7.86 (s, 1H),7.80 (s, 1H), 5.32 (t, 2H, J =4.2 Hz), 4.93-4.74 (m, 5H), 4.54-4.37 (m, 3H), 4.33 (t, 2H, J =7.0 Hz), 4.21, 4.14 (ABq, 2H, J =12.1 Hz), 3.95-3.48 (m, 26H),3.37 (t, 1H, J=9.6 Hz), 3.31-3.19 (m, 2H), 2.99-2.90 (m, 1H), 2.69 (t, 2H, J =7.4 Hz), 1.86-1.76 (m, 2H), 1.61-1.50 (m, 2H), 1.29-0.83 (m, 12H), 0.79 (t,3H, J =7.0 Hz); 13 C NMR (D2O, 100 MHz): δ =147.7, 143.4, 142.8, 125.9, 125.1,124.2, 101.3, 99.9, 99.8, 99.6, 99.0, 77.2, 77.0, 76.1, 74.6, 73.3, 72.9 (×2), 72.7, 71.8 (×2), 71.5, 71.2, 70.6, 70.5, 70.0, 69.8, 69.3, 68.1 (×2),67.9, 61.9, 60.7, 60.5, 58.9, 51.3, 51.1, 50.0, 31.0, 28.6, 28.4, 28.3, 28.2, 25.5, 25.3, 23.8, 22.1, 13.5.

[0671] Synthesis of Compound 20

[0672] 20

[0674] The deprotected oligosaccharide 19 (55 mg, 0.04 mmol) was sulfated according to general procedure 2 and then dialyzed to obtain oligosaccharide 20 (30 mg, 26%) as a white powder. 1 H NMR (D2O, 400 MHz): δ =8.16 (s, 1H), 8.02(s, 1H), 8.00 (s, 1H), 5.75-5.66 (m, 1H), 5.64-5.55 (m, 1H), 8.26 (s, 1H),5.23-4.88 (m, 10H), 4.88-4.66 (m), 4.66-4.31 (m, 15H), 4.30-4.12 (m, 5H), 4.06 (t, 1H, J=10.0 Hz), 3.45-3.36 (m, 1H), 2.06-1.90 (m, 2H), 1.78-1.62 (m,2H), 1.44-0.96 (m, 10H), 0.89 (t, 3H, J =7.0 Hz); 13 C NMR (CDCl3, 100 MHz): δ =125.9, 125.6, 124.3, 99.4, 97.1, 96.3, 94.3, 93.6, 76.3, 75.5, 75.2, 74.7,73.7, 73.5, 73.0, 72.7, 72.6, 71.6, 71.4, 70.7, 70.6, 69.8, 67.8, 66.5, 65.9,62.1, 59.8, 51.1, 50.9, 50.1, 31.0, 28.8, 28.4, 28.2, 28.1, 25.6, 25.2, 24.0, 22.1, 13.5.

[0675] Example 10 - Synthesis of Compound 25 (g19)

[0676] 25

[0678] The general synthetic scheme for compound 25 is as follows.

[0679]

[0680]

[0681] Program 3 for synthesizing SO3.Py

[0682] Chlorosulfonic acid (1.6 mL) was added dropwise to a solution of pyridine (4 mL) in DCM (20 mL) at 0 °C, and the mixture was stirred for 30 min. The reaction mixture was then filtered, and the precipitate was washed with ice-cold water (3 × 20 mL), ice-cold saturated NaHCO3 (2 × 10 mL), ice-cold water (2 × 10 mL) (until the pH of the filtrate was 7), cold EtOH (2 × 20 mL), cold toluene (10 mL), and cold DCM (10 mL). The precipitated SO3·Py was then thoroughly dried under high vacuum and used within 24 h.

[0683] General Program 3 for Click Response

[0684] Sodium ascorbate (2 equivalents) and copper sulfate pentahydrate (1 equivalent) were added to a mixture of an azide analog (1 equivalent) and an alkyne analog (1 equivalent / N3) in 1,4-dioxane / H2O (3:1). The reaction mixture was then stirred at room temperature for 18 h and filtered on a diatomaceous earth mat. The filtrate was concentrated under reduced pressure, and the crude substance was dissolved in EtOAc. The organic layer was washed with 1M HCl and saturated NaHCO3, dried (MgSO4), and concentrated under reduced pressure. The resulting crude substance was purified by rapid column chromatography on silica gel to obtain the desired compound.

[0685] General Procedure 3 for Acetate Hydrolysis

[0686] A solution of 30% NH3 aqueous solution (3-5 mL) and oligosaccharide in methanol (20 mL) was stirred at room temperature for 48 h. The reaction mixture was concentrated under reduced pressure, and the crude material was dissolved in ultrapure water (5-10 mL) and washed with EtOAc (5 × 10 mL). The aqueous layer was concentrated to obtain the pure compound.

[0687] General procedure for sulfation 3

[0688] Freshly prepared SO3·Py (10 equivalents / OH) complex was added to a solution of starting material in dry DMF (10 mL) maintained under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 2 days and adjusted to alkalinity with 5 M NaOH (2 equivalents / SO3·Py). The mixture was then concentrated under reduced pressure, and the crude material was dissolved in ultrapure water (10–20 mL) and dialyzed according to a standard procedure.

[0689] General Procedure for Dialysis 3

[0690] Commercially available cellulose membrane dialysis tubing with MWCO at 0.5 kD, 1 kD, and 2 kD was used for dialysis. Before use, the tubing was washed with milliQ water (for approximately 5–10 minutes). One end of the tubing was tied, and the sample was loaded into the tubing. The tubing was then clamped shut, leaving almost no space for air bubbles. A 3 L flask equipped with a stir bar was filled with milliQ water and placed on a magnetic stir plate at room temperature. The stirring was set so that the dialysis bag floated slowly on top of the solution in the flask. After 2 hours, the water was replaced with fresh milliQ water, and dialysis was continued for 2 days, with the dialysis water replaced every 18 hours. The dialysis bag was then removed from the flask, and its contents were collected and freeze-dried.

[0691] Synthesis of Compound 21

[0692] twenty one

[0694] By performing a click reaction of sucralose azide 2 (prepared according to Example 2) (2.05 g, 3.33 mmol) and 4-pentyn-1-ol (430 mg, 5.1 mmol) using general procedure 3, compound 21 (2.00 g, 86%) was obtained. 1 HNMR (CDCl3, 400 MHz): δ =7.47 (s, 1H, H-7'), 5.68-5.64 (m, 2H, H-1, H-3'), 5.36 (t, 1H, J =7.2 Hz, H-4), 5.30-5.29 (m, 2H, H-4', H-3), 4.76 (part A of ABX, 1H, J 5,6a =3.6 Hz, J 6a,6b =14.4 Hz, H-6a), 4.63 (part B of ABX, J 5,6b =8.7 Hz, J 6a,6b =14.4 Hz, H-6b), 4.60-4.54 (m, 2H, H-2, H-5'), 4.41-4.34 (m, 1H, H-5), 4.28 (part A of ABX, J 5’,6’a =4.9 Hz, J 6’a,6’b =11.8 Hz, H-6'a), 4.21 (part B of ABX, J 5’,6’b =6.7 Hz, J 6’a,6’b =11.8 Hz, H-6'b), 3.67 (t, 2H, J 10’,11’ =6.1 Hz, H-11'), 3.61, 3.47(ABq, 2H, J =12.0 Hz, H-1'), 2.82 (t, 2H, J 9’,10’ =7.3 Hz, H-9'), 2.11 (s, 3H,OCH3), 2.10 (s, 3H, OCH3), 2.07 (s, 6H, 2×OCH3), 2.04 (s, 3H, OCH3), 1.96-1.87 (m, 2H, H-10'); 13C NMR (CDCl3, 100 MHz): δ =170.4, 170.2, 170.2 (×2),169.6, 147.5 (C-8'), 122.6 (C-7'), 103.9 (C-2'), 90.4 (C-1), 79.4 (C-5), 75.3(C-4), 75.2 (C-2'), 68.1, 67.7, 67.0, 63.8 (C-6'), 61.5 (C-11'), 58.9, 52.4(C-6), 44.8 (C-9'), 31.7 (C-10'), 21.8, 20.7 (×2), 20.6 (OCH3), 20.5 (OCH3),20.4 (OCH3).

[0695] Synthesis of Compound 22

[0696] twenty two

[0698] Triethylamine (1.5 mL) and methanesulfonyl chloride (0.5 mL, 4.76 mmol) were added to a solution of ethanol 21 (2.00 g, 2.86 mmol) in anhydrous DCM (20 mL), and the mixture was stirred at room temperature for 18 h. The reaction mixture was washed with 1 M HCl (2 × 30 mL) and saturated NaHCO3 (30 mL), dried to MgSO4, and concentrated under reduced pressure. NaN3 (304 mg, 4.76 mmol) was added to a solution of crude material in anhydrous DMF (20 mL), and the mixture was stirred at 60 °C for 18 h. The reaction mixture was concentrated under reduced pressure, and the crude material was dissolved in EtOAc (100 mL), washed with 1 M HCl (50 mL) and saturated NaHCO3 (50 mL), dried to MgSO4, and concentrated under reduced pressure. The crude material was purified by rapid column chromatography (1.5:1 EtOAc / Pet.Sp. to EtOAc) to give azide 22 (1.24 g, 60%). 1 H NMR (CDCl3, 400 MHz): δ =7.43 (s, 1H, H-7'), 5.65-5.61 (m, 2H, H-1, H-3'), 5.34 (t, 1H, J =7.2 Hz, H-4), 5.29-5.25 (2H, H-3, H-4'), 4.74 (part A of ABX, 1H, J 5,6a =3.6 Hz, J6a,6b =14.3Hz, H-6a), 4.60 (B part of ABX, J 5,6b =8.8 Hz, J 6a,6b =14.4 Hz, H-6b), 4.56-4.51 (m,2H, H-2, H-5'), 4.38-4.31 (m, 1H,H-5),), 4.25 (part A of ABX, J 5’,6’a =4.8 Hz, J 6’a,6’b =11.8 Hz, H-6'a), 4.17 (part B of ABX, J 5’,6’b =6.7 Hz, J 6’a,6’b =11.8 Hz, H-6'b), 3.58, 3.44 (ABq, 2H, J =12.0 Hz, H-1'), 3.30 (t, 2H, J 10’,11’ =6.7 Hz, H-11'), 2.76 (t, 2H, J 9’,10’ =7.4 Hz, H-9'), 2.08 (s, 3H, OCH3), 2.07 (s, 3H,OCH3), 2.03 (s, 6H, 2×OCH3), 2.01 (s, 3H, OCH3), 1.98-1.91 (m, 2H, H-10'); 13 CNMR (CDCl3, 100 MHz): δ =170.2, 169.9 (×3), 169.5, 146.5, 122.3, 103.8,90.3, 79.4, 75.2, 75.1, 68.0, 67.6, 66.8, 63.7, 58.8, 52.2, 50.5, 44.8, 28.3,22.4, 20.6 (×2), 20.4, 20.3.

[0699] Synthesis of Compound 23

[0700] twenty three

[0702] A click reaction of azide 22 (500 mg, 0.70 mmol) and propargyl maltotriose 11 (prepared according to Example 7) (700 mg, 0.70 mmol) was carried out using general procedure 3 to obtain oligosaccharide 23 (700 mg, 59%) as a glassy solid. 1 H NMR (CDCl3, 400 MHz): δ =7.57 (s, 1H), 7.47 (s, 1H), 5.67-5.60 (m,2H), 5.36-5.21 (m, 6H), 5.21-5.12 (m, 2H), 4.97 (t, 1H, J =10.0 Hz), 4.86-4.68(m, 5H), 4.68-4.49 (m, 5H), 4.46-4.28 (m, 5H), 4.23 (dd, 2H, J =4.4 Hz, J =12.0Hz), 4.19-4.07 (m, 3H), 3.96 (dd, 1H, J =2.3 Hz, J =12.4 Hz), 3.93-3.81 (m,4H), 3.73-3.62 (m, 1H), 3.57, 3.47 (ABq, 2H, J =12.1 Hz), 2.73-2.57 (m, 2H),2.24 (t, 2H, J =6.7 Hz), 2.08-1.87 (15×s, 45H); 13C NMR (CDCl3, 100 MHz): δ =170.8, 170.4, 170.3 (×2), 170.1 (×2), 169.9, 169.8, 169.6, 169.5, 169.4 (×2), 169.2, 147.9, 145.7, 143.8, 123.0, 122.8, 103.9, 99.2, 95.5, 95.4, 90.4,79.4, 75.2, 75.0, 73.6, 72.4, 72.0, 71.8, 71.5, 70.2, 69.8, 69.1, 68.7, 68.3,68.0, 67.7, 67.6, 66.7, 63.8, 62.7, 62.6, 62.1, 61.2, 58.8, 52.3, 48.9, 44.6, 29.4, 21.8, 20.8, 20.7, 20.6 (×2), 20.5 (×2), 20.4, 20.3 (×2), 20.2.

[0703] Synthesis of Compound 24

[0704] twenty four

[0706] Compound 23 (700 mg, 0.42 mmol) was hydrolyzed with acetate according to general procedure 3 to obtain oligosaccharide 24 (412 mg, 93%). 1 H NMR (D2O, 400 MHz): δ =8.02 (s, 1H), 7.79 (s, 1H), 5.43-5.38(m, 3H), 4.98, 4.85 (ABq, 2H, J =12.6 Hz), 4.68 (d, 1H, J =7.9 Hz), 4.54 (d,1H, J =3.8 Hz), 4.51-4.40 (m, 4H), 4.24-4.15 (m, 2H), 4.04-3.55 (m, 21H), 3.44(t, 1H, J =9.3 Hz), 3.37 (dd, 1H, J =1.3 Hz, J =9.4 Hz, 1H), 2.73 (t, 2H, J =7.1Hz), 2.31 (p, 2H, J =7.2 Hz,J =13.8 Hz); 13 C NMR (D2O, 100 MHz): δ =177.3,146.6, 143.5, 125.2, 124.1, 103.4, 101.9, 101.4, 99.8, 99.6, 99.3, 92.4,79.5, 78.9, 77.1, 76.9, 76.1, 75.6, 74.8, 74.6, 73.3, 72.9 (×2), 72.7, 71.8,71.5, 71.2, 70.6, 69.3, 68.0, 67.5, 63.1, 62.0, 61.8, 60.7, 60.5, 52.1, 49.6, 43.8, 28.7, 21.6.

[0707] Synthesis of Compound 25

[0708] 25

[0710] The deprotected pentamer 24 (235 mg, 0.22 mmol) was sulfated according to general procedure 3 and then dialyzed to give pentamer 25 (102 mg, 17%) as a white powder. 1 H NMR (D2O, 400 MHz): δ =8.21 (s, 1H),7.98 (s, 1H), 5.83 (d, 1H, J =3.4 Hz), 5.76-5.69 (m, 1H), 5.63 (d, 1H, J =2.9Hz), 5.42 (d, 1H, J =7.4 Hz), 5.21-4.87 (m, 11H), 4.93-4.61 (m, 7H), 4.62-4.14(m, 17H), 4.04, 3.95 (ABq, 2H, J =12.1 Hz), 2.47-2.29 (m, 2H); 13C NMR (CDCl3,100 MHz): δ =125.7, 124.7, 103.6, 99.4, 94.4, 93.7, 90.9, 79.5 (×2), 78.3,76.3, 76.1, 74.7, 73.6, 73.0, 72.6, 72.4, 71.8, 71.6, 71.4, 70.2, 69.8, 68.4,67.8, 66.5, 66.0, 62.0, 60.3, 52.7, 49.7, 44.0, 29.0, 21.6.

[0711] Example 11 - Synthesis of Compound 29 (G4)

[0712] 29

[0714] Program 4 for synthesizing SO3.Py

[0715] Chlorosulfonic acid (1.6 mL) was added dropwise to a solution of pyridine (4 mL) in DCM (20 mL) at 0 °C, and the mixture was stirred for 30 min. The reaction mixture was then filtered, and the precipitate was washed with ice-cold water (3 × 20 mL), ice-cold saturated NaHCO3 (2 × 10 mL), ice-cold water (2 × 10 mL) (until the pH of the filtrate was 7), cold EtOH (2 × 20 mL), cold toluene (10 mL), and cold DCM (10 mL). The precipitated SO3·Py was then thoroughly dried under high vacuum and used within 24 h.

[0716] General Procedure 4 for Sulfation

[0717] Freshly prepared SO3·Py (10 equivalents / OH) complex was added to a solution of starting material in dry DMF (10 mL) maintained under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 2 days. The DMF was then decanted from the reaction mixture, and the precipitate was dissolved in ultrapure water (10 mL) and adjusted to alkalinity (pH 9-11) using 5 M NaOH. The solution was then dialyzed according to a standard procedure.

[0718] General Procedure for Dialysis 4

[0719] Commercially available cellulose membrane dialysis tubing with MWCO at 0.5 kD, 1 kD, and 2 kD was used for dialysis. Before use, the tubing was washed with milliQ water (for approximately 5–10 minutes). One end of the tubing was tied, and the sample was loaded into the tubing. The tubing was then clamped shut, leaving almost no space for air bubbles. A 3 L flask equipped with a stir bar was filled with milliQ water and placed on a magnetic stir plate at room temperature. The stirring was set so that the dialysis bag floated slowly on top of the solution in the flask. After 2 hours, the water was replaced with fresh milliQ water, and dialysis was continued for 2 days, with the dialysis water replaced every 18 hours. The dialysis bag was then removed from the flask, and its contents were collected and freeze-dried.

[0720] General Procedure 4 for the Hydrolysis of Acetates and Benzoyl Esters

[0721] A solution of 30% NH3 aqueous solution (3-5 mL) and oligosaccharide in methanol (20 mL) was stirred at room temperature for 48 h. The reaction mixture was concentrated under reduced pressure, and the crude material was dissolved in ultrapure water (5-10 mL) and washed with EtOAc (5 × 10 mL). The aqueous layer was concentrated to obtain the pure compound.

[0722] General Procedure 4 for Click Response

[0723]

[0724] Sodium ascorbate (2 equivalents) and copper sulfate pentahydrate (1 equivalent) were added to a mixture of an azide analog (1 equivalent) and an alkyne analog (1 equivalent / N3) in 1,4-dioxane / H2O (3:1). The reaction mixture was then stirred at room temperature for 18 h and filtered on a diatomaceous earth mat. The filtrate was concentrated under reduced pressure, and the crude substance was dissolved in EtOAc. The organic layer was washed with 1M HCl and saturated NaHCO3, dried (MgSO4), and concentrated under reduced pressure. The resulting crude substance was purified by rapid column chromatography on silica gel to obtain the desired compound.

[0725] Synthesis of acetylated propargyl acarbose (compound 26)

[0726]

[0727] Boron trifluoride diethyl ether (0.6 mL, 5 mmol) was added dropwise to a mixture of peracetic acarbose (0.6 g, 0.5 mmol) and propargyl alcohol (35 μL, 0.6 mmol) in a dry DCM (20 mL) at 0 °C. The mixture was then stirred at room temperature for 5 h. Saturated NaHCO3 (10 mL) was then added to the reaction mixture and stirred for another 10 min. The organic layer was extracted, dried (MgSO4), and concentrated under reduced pressure. The resulting crude product was purified by rapid column chromatography (3:1 EtOAc / Pet. Sp.) to give propargyl acarbose 26 (0.1 g, 17%) as a glassy solid. 1 H NMR (CDCl3, 400 MHz): δ =5.95 (d, 1H, J =5.3 Hz), 5.60-5.51 (m, 2H), 5.40-5.31 (m,1H), 5.31-5.18 (m, 3H), 5.10 (t, 1H, J =10.2 Hz), 4.92 (dd, 1H, J =4.3 Hz, J =10.0 Hz), 4.85-4.69 (m, 4H), 4.67-4.60 (m, 1H), 4.49 (dd, 1H, J =2.9 Hz, J =12.3 Hz), 4.47-4.34 (d, 2H), 4.33 (d, 2H, J =2.3 Hz), 4.29 (dd, 1H, J =4.0 Hz, J =12.4 Hz), 4.20-4.14 (m, 1H), 3.99 (t, 1H, J =9.3 Hz), 3.94-3.88 (m, 2H), 3.77-3.67 (m, 2H), 3.57-3.46 (m, 1H), 2.46 (t, 1H, J =2.4 Hz), 2.39 (t, 1H, J =9.9 Hz), 2.16, 2.13, 2.09, 2.04, 2.02, 2.01, 2.00, 1.97, 1.96 (s, OCH3) ppm; 13C NMR (CDCl3, 100 MHz): δ =171.0, 170.7, 170.6, 170.5, 170.3 (×2), 170.2,169.9, 169.7, 169.6, 133.9, 128.0, 97.5, 95.8, 95.6, 78.0, 77.2, 75.5, 75.3,73.3, 72.2 (×2), 71.8 (×2), 71.0, 70.9, 70.7, 70.5, 70.1, 69.8, 69.1, 63.0,62.7, 62.2, 61.1, 55.8, 52.1, 20.8, 20.7, 20.6, 20.5 (×2), 18.1 ppm.

[0728] Synthesis of triethylene ether-linked diacarbose (compound 27)

[0729] 27

[0731] Through general procedures 4. A click reaction was carried out between propargyl acarbose 26 (350 mg, 0.30 mmol) and 1,2-bis(2-azidoethoxy)ethane (30 mg, 0.13 mmol) to give diacarbose 27 (235 mg, 71%) as a solid. 1 H NMR (CDCl3, 400 MHz): δ =7.62 (s, 2H), 5.93 (d, 2H, J =5.2 Hz), 5.59-5.49 (m, 4H), 5.37-5.29 (m, 2H), 5.26-5.17 (m, 6H), 5.07 (t, 2H, J =10.2 Hz),4.93-4.86 (m, 4H), 4.83-4.58 (, 12H), 4.56-4.40 (m, 8H), 4.38-4.22 (m, 4H), 4.19-4.11 (m, 2H), 4.00-3.79 (m. 10H), 3.74-3.65 (m, 4H), 3.54-3.44 (m, 4H), 2.36 (t, 2H, J =10.0 Hz), 2.14 (s, 6H), 2.12 (s, 6H), 2.10 (s, 6H), 2.04-1.91(m, 54H), 1.18 (d, 6H, J =6.2 Hz);13 C NMR (CDCl3, 100 MHz): δ =171.0, 170.7,170.6, 170.4, 170.3 (×2), 170.2, 170.0, 169.9, 169.7, 169.6, 143.8, 133.9,128.0, 124.0, 99.44, 95.8, 95.6, 77.2, 75.3, 73.4, 72.2 (×2), 72.0, 71.9,71.0, 70.9, 70.7, 70.5, 70.4, 70.1, 69.8, 69.4, 69.1, 63.0, 62.9, 62.7, 62.2, 61.1, 52.1, 50.2, 20.9, 20.8, 20.7, 20.6 (×2), 20.5, 18.1.

[0732] Synthesis of diacarbose (compound 28)

[0733] 28

[0735] Compound 27 (0.24 g, 94 μmol) was hydrolyzed with acetate according to general procedure 4 to give diacarbose compound 28 (0.15 mg, 99%). 1 H NMR (D2O, 400 MHz): δ =8.00 (s, 1H), 5.86-5.82(m, 1H), 5.32 (d, 1H, J =3.8 Hz), 5.25 (d, 1H, J =3.4 Hz), 4.92, 4.80 (ABquat.2H), 4.56-4.47 (m, 2H), 4.17, 4.06 (ABquat., 2H, J =13.7 Hz), 3.98 (d, 1H, J =6.9 Hz), 3.91-3.43 (m, 22H), 3.26 (dd, 1H, J =8.0 Hz, J =9.4 Hz), 2.41 (t, 1H, J =9.5 Hz), 1.28 (d, 3H, J =6.3 Hz); 13C NMR (D2O, 100 MHz): δ =143.5, 138.9,125.7, 123.8, 101.3, 100.0, 99.5, 77.1, 77.0, 74.6, 73.3, 73.0, 72.9 (×2),72.7, 71.4, 71.2 (×2), 70.8, 69.6, 68.7, 64.9, 61.9, 61.6, 60.7, 60.5, 56.0,50.1, 17.3.

[0736] Synthesis of sulfated diacarbose (compound 29)

[0737] 29

[0739] The deprotected octamer 28 (50 mg, 0.03 mmol) was sulfated according to general procedure 4, followed by dialyzing, to give 29 (60 mg, 53%) as a white powder; 1 H NMR (D2O, 400 MHz): δ =8.18 (s, 2H), 6.14(s, 2H), 5.67 (d, 2H, J =3.1 Hz), 5.59 (d, 2H, J =2.7 Hz), 5.57-5.52 (m, 2H), 5.22-5.14 (m, 2H), 5.14-5.55 (m), 4.55-4.38 (m, 6H), 4.37-4.18 (m, 8H), 4.18-4.04 (m, 4H), 4.02-3.90 (m 4H), 3.74-3.57 (m, 4H), 3.45-3.27 (m, 2H), 1.54(d, 6H, J =6.0 Hz).

[0740] Example 12 - Synthesis of Compound 30 (G17)

[0741] Compound 30 (G17) was prepared according to the procedure disclosed in WO 2018 / 068090 A1 (the entire contents of which are incorporated herein by reference).

[0742] 30

[0744] Example 13 – Inhibition of the interaction between heparin and the SARS-CoV-2 spike protein and its receptor-binding domain (RBD)

[0745] The ability of compounds to inhibit the interaction between heparin and the SARS-CoV-2 spike protein and its receptor-binding domain (RBD) was evaluated using surface plasmon resonance (SPR). The optical phenomena of SPR were used to monitor the physical interactions between molecules. A solution of a potential protein ligand (e.g., a viral protein such as the spike protein from SARS-CoV-2, or its RBD) was flowed through a sensor surface coupled to a target (e.g., heparin) to monitor the real-time binding of the protein ligand to the immobilized target. Detection was achieved by measuring changes in refractive index very close to the sensor surface. As the refractive index changes, the angle at which plasmon resonance occurs also changes, and this change is directly related to the amount of protein interacting with the surface. The BIAcore T200 was conveniently used. It is highly sensitive, and its microfluidic technology ensures that only a small amount of material is required.

[0746] For this experiment, biotinylated heparin was immobilized on a biosensor chip. Biotinylation occurs via a reducing end modified with an amino group or by ammonia reduction amination, using sulfonyl-NHS-biotin (see, for example, Osmond et al. (2002)). Anal Biochem , 310(2):199-207). A solution containing the potential target protein ligand was injected onto the surface of the sensor chip and the binding was measured in real time (reference, e.g., Fernig (2001) see: Proteoglycan protocols, RV Iozzo ed., Humana Press, Totowa, NJ, USA).

[0747] To screen for sulfated glycoconjugates, a competitive inhibition assay was used. In this case, the test compound (G24, G17, G4, G19, or G22, prepared according to Examples 1-12) was mixed with the target protein (e.g., the full-length spike protein or its RBD) in the liquid phase, and then the mixture was flowed through the surface of a biosensor chip already conjugated with heparin. If the test compound binds to the protein in a manner that inhibits the subsequent binding of the protein to heparin, a reduced signal is detected compared to the absence of the test compound. The IC50 was calculated by titrating the test compound while maintaining a constant protein concentration. 50 Then you can use IC 50 To compare the inhibition of protein binding by various test compounds. The protein concentration used in this assay varied depending on the protein being detected and was determined based on the signal obtained in the absence of the test compound. The concentration of RBD used for the spike protein was 120 nM, and the concentration used for the full-length spike protein was 95 nM.

[0748] The results are shown in Table 1.

[0749] Table 1: Inhibition of the interaction between the full-length SARS-CoV-2 spike protein and its RBD.

[0750]

[0751] Example 14 – The ability of G17, G22, and G24 to inhibit the binding of RBD to Calu-3 cells expressing ACE2

[0752] Calu-3, a human adenocarcinoma cell line expressing the human ACE2 receptor, was used. Calu-3 cells (25 x 10⁻⁶ cells) were cultured in Eagle's minimum essential medium (MEM) containing 10% fetal bovine serum. 4 Cells were seeded into the wells of a 24-well plate containing collagen-coated coverslips. Cells were allowed to adhere to the coverslips for approximately 1 h, and then the volume of culture medium in the wells was increased to 1 mL. Culture was continued for 5 days to ensure adequate cell coverage of the coverslips. Recombinant SARS-CoV-2 spike protein receptor-binding domain (RBD) with a C-terminal His tag (9.4 μg / mL) was mixed with heparin (100 μg / mL) or the test compound (G17, G22, or G24, prepared according to Examples 6, 9, and 12; 100 μg / mL) and incubated at room temperature for 45 min. All concentrations were final, and the basal medium was Eagle's minimum essential medium (MEM). The mixture was then added to a Calu-3 cell layer and incubated at 37 °C for 60 min, followed by fixation with 4% paraformaldehyde at 37 °C for 15 min. Cells were washed with phosphate-buffered saline (PBS) and blocked overnight at 4 °C with a solution of 1% bovine serum albumin (BSA) in PBS. They were then washed in PBS, incubated with 1 μg / mL anti-His antibody, and then incubated with AlexaFlour 568-conjugated anti-mouse secondary antibody at 37°C for 60 min. The coverslips were washed and fixed, and observed using a Nikon confocal microscope.

[0753] The results show Figure 1 The study confirmed that G22 is the most effective inhibitor of RBD binding to Calu-3 cells. Heparin and G24 also inhibited RBD binding to Calu-3 cells, but G17 did not show significant inhibition.

[0754] Example 15 - The ability of G4, G17, G19, G22 and G24 to inhibit SARS-CoV-2 virus infection in VERO E6 monkey kidney cells

[0755] The cell line was obtained from the European Collection of Authenticated Cell Cultures (ECACC), UK Health Security Agency (UKHSA), Porton Down, United Kingdom. SARS-CoV-2 (hCoV-19 / Australia / VIC01 / 2020) (Victoria isolate) was provided by the Doherty Institute in Melbourne, Australia, at passage P1, and passaged twice in Vero / hSLAM cells [ECACC04091501]. Whole-genome sequencing of the third-generation working stock was performed using Nanopore and Illumina technologies, and no significant changes in the viral sequence were observed. Viral titers were determined by a focus-forming assay on Vero-E6 cells [ECACC 85020206]. Cell cultures were maintained at 37°C in minimally essential medium (MEM) (Life Technologies, California, USA), supplemented with 10% fetal bovine serum (FBS) (Sigma, Dorset, UK) and 25 mM 4-(2-hydroxyethyl)-1-piperazine ethanesulfonic acid (HEPES; Life Technologies, California, USA), 2 mM L-glutamine (Gibco, Thermo Fisher Scientific Inc., Waltham, MA, USA) and 1x non-essential amino acid solution (Gibco, Thermo Fisher Scientific Inc., Waltham, MA, USA). Additionally, Vero / hSLAM cultures were supplemented with 0.4 mg / mL of genimycin (Invitrogen, Thermo Fisher Scientific Inc., Waltham, MA, USA) to maintain stable integration of pCAG-hSLAM and expression of human signaling lymphocyte activation molecule (hSLAM). At UKHSA, PortonDown, different variants of interest were cultured from clinical samples in a manner similar to that of the Victoria isolate.

[0756] Vero E6 monkey kidney cells (EACC 85020206) were seeded into 96-well plates. After 24 hours, the test compounds [G4, G17, G19, G22, G24 (prepared according to Examples 1-12) or unfractionated heparin] were serially diluted, and each dilution was mixed with a fixed number of PFU (e.g., 70) of SARS-CoV-2 virus (hCoV-19 / Australia / VIC01 / 2020) (50:50) and incubated in 96-well plates at 37°C for 1 hour. The virus / compound mixture was added to the monolayer of cells, and the virus was allowed to adsorb at 37°C for 1 hour. After 1 hour, the mixture was removed, and covering medium containing the diluted compounds was added to each well. The plates were incubated at 37°C for 24 hours. Cells were fixed with 8% formalin for at least 8 hours, and then the fixed cells were subjected to an immunostaining protocol using an anti-spike protein antibody. The stained lesions were counted using an ELISpot counter (Cellular Technology Limited (CTL)). The counted lesion data were then imported into R-Bioconductor. On each assay plate, a positive control chloroquine biphosphate (50-0.02 μM) was run along with the test compound.

[0757] The results are shown in Table 2 and Figure 2 As shown in (Run 1) and 3 (Run 2).

[0758] Table 2: Inhibition of viral infection in VERO E6 monkey kidney cells.

[0759]

[0760] The ability of G24 to inhibit different SARS-CoV-2 variants was evaluated using the above protocol. Briefly, VeroE6 monkey kidney cells (EACC 85020206) were seeded into 96-well plates. After 24 hours, G24 (prepared according to Example 6) was serially diluted, and each dilution was mixed with a fixed number of PFU (e.g., 70) of SARS-CoV-2 virus [hCoV-19 / Australia / VIC01 / 2020 (Victoria, Wuhan-like isolate), Delta variant, Omicron variant, or Beta variant] (50:50) and incubated in 96-well plates at 37°C for 1 hour. The virus / compound mixture was added to a VeroE6 monkey kidney cell monolayer, and the virus was allowed to adsorb onto the cells at 37°C for 1 hour. After 1 hour, the mixture was removed, and a covering medium (1% carboxymethyl cellulose) containing the diluted compound was added to each well. Depending on the variant, the plates were incubated at 37°C for 20–26 hours. Cells were fixed in 8% formalin for at least 8 hours, and then subjected to an immunostaining protocol. For assays involving Omicron variants, the following procedure was performed: After formalin fixation, cells were washed and permeabilized for 10 minutes at room temperature with 0.2% (w / v) Triton X-100 / PBS, followed by washing with PBS, incubating with 0.3% hydrogen peroxide at room temperature for 20 minutes, and washing with PBS. Lesions were stained with rabbit anti-nucleocapsid antibody (diluted in 0.2% (w / v) Triton X-100 / PBS) at room temperature for 1 hour, followed by washing and staining with goat anti-rabbit immunoglobulin antibody (conjugated with horseradish peroxidase (HRP), diluted in the same buffer). After washing, cells were incubated with TrueBlue peroxidase substrate (SeraCare LifeSciences Inc., Milford, Massachusetts, USA) at room temperature for 10 minutes and then washed to complete the procedure. The stained lesions were counted using an ELISpot counter (Cellular Technology Limited (CTL)). The counted lesion data were then imported into R-Bioconductor. On each assay plate, a positive control chloroquine hydrogen phosphate (50–0.02 μM) was run along with the test compound. Midpoint probability analysis (written in R for statistical calculations and graphing) was used to determine 50% (IC50) inhibition of SARS-CoV-2 infectious viral lesions compared to the virus-only control. 50 The required amount of compound (μg / mL).

[0761] The results are shown in Table 3.

[0762] Table 3: G24 inhibits viral infection of VERO E6 monkey kidney cells by SARS-CoV-2 variants.

[0763]

[0764] Example 16 - Ability of G19, G22 and G24 to inhibit IL-8 (CXCL8)-dependent chemokines

[0765] These experiments were performed using DMSO-treated human promyelocytic HL-60 cells. These cells were derived from patients with acute promyelocytic leukemia. Cells were treated with DMSO (1.2%) for 4 days prior to use in the experiments. Chemotaxis assays were performed in 96-well Costa chemotaxis plates consisting of a bottom chamber containing human IL-8, with or without G19, G22, or G24 (prepared according to Examples 6, 9, and 10; 50 μg / mL). Cells in RPMI and 1% v / v FCS were added to the upper chamber, and the plate was incubated at 37°C for 1 hour to allow cell migration from the upper chamber to the lower chamber. The number of cells migrating to the lower chamber was quantified by labeling with AQUEOUS ONE (20 μL / well) for 1.75 hours, followed by absorbance readings at 490 nm. IL-8 was used at a final concentration of 20 ng / mL. Mean, upper limit, and lower limit values ​​are shown.

[0766] The results are as follows Figure 4 As shown in the figure, the percentage of inhibition is displayed in the presence of G19, G22, or G24. All three compounds inhibited IL-8-dependent chemokines, with G24 causing the greatest inhibition.

[0767] The publications of each patent, patent application, and publication cited in this article are hereby incorporated in their entirety by reference.

[0768] Any reference cited herein should not be construed as an admission that such reference is available as “background art” for this application.

[0769] This specification is intended to describe preferred embodiments of the invention and not to limit the invention to any particular set of embodiments or features. Therefore, those skilled in the art will recognize that various modifications and variations can be made to the specific embodiments described herein without departing from the scope of the invention. All such modifications and variations are intended to be included within the scope of the appended claims.

Claims

1. Compounds of formula I, II, or III: (I) (II) (III) Or its pharmaceutically acceptable salt, solvate, or prodrug, in: L 1 L 3 L 4 L 6 L 7 L 8 L 9 L 10 L 11 L 12 L 13 L 15 L 16 L 17 L 18 and L 19 Independently selected from optionally substituted C1-C5 alkylene groups, optionally substituted C2-C5 alkenyl groups, and optionally substituted C2-C5 ynylene groups; L 2 L 5 and L 14 Independently selected from optionally substituted C2-C8 alkylene groups, optionally substituted C2-C8 alkenyl groups, and optionally substituted C2-C8 ynylene groups; X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 and X 11 Independently selected from CH and N; and R 1 Selected from C2-C which are optionally replaced 12 Alkyl groups, optionally substituted C2-C 12 Alkenyl and optionally substituted C2-C 12 Alkyne group.

2. The compound according to claim 1, wherein X 1 -X 11 It is CH.

3. The compound according to claim 1 or claim 2, wherein L 1 L 3 L 4 L 6 L 7 L 8 L 9 L 10 L 11 L 12 L 13 L 15 L 16 L 17 L 18 and L 19 It is a C1-C3 alkylene group that is optionally substituted.

4. The compound according to claim 3, wherein L 1 L 3 L 4 L 6 L 7 L 8 L 9 L 10 L 11 L 12 L 13 L 15 L 16 L 17 L 18 and L 19 It is methylene.

5. The compound according to any one of claims 1-4, wherein L 2 It is an optional substituted C2-C5 alkylene group.

6. The compound according to claim 5, wherein L 2 It is propylidene.

7. The compound according to any one of claims 1-6, wherein L 5 It is a C3-C6 alkylene group that is optionally substituted.

8. The compound according to claim 7, wherein L 5 It is Adinaki.

9. The compound according to any one of claims 1-8, wherein L 14 It is a C2-C4 alkylene group that is optionally substituted.

10. The compound according to claim 9, wherein L 14 It is ethylene.

11. The compound according to any one of claims 1-10, wherein R 1 C6-C is arbitrarily replaced. 10 alkyl.

12. The compound according to claim 11, wherein R 1 It's Hsinki.

13. The compound according to any one of claims 1-12, wherein the compound is a compound of formula IV, V or VI or a pharmaceutically acceptable salt, solvate or prodrug thereof: (IV), (V), or (VI).

14. The compound according to any one of claims 1-13, wherein the compound is in the form of a salt and the salt is a sodium salt.

15. A pharmaceutical composition comprising a compound according to any one of claims 1-14 or a pharmaceutically acceptable salt, solvate or prodrug thereof, and a pharmaceutically acceptable carrier or diluent.

16. The compound according to any one of claims 1-14, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, for use in a therapeutic manner.

17. A method for treating or inhibiting the development of a viral infection in a subject, said method comprising, consisting of, or substantially consisting of: The subject is given a compound according to any one of claims 1-14, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, or a compound of formula VII, or a pharmaceutically acceptable salt, solvate, or prodrug thereof: (VII) 18. The method of claim 17, wherein the infection is caused by a virus expressing a viral fusion protein.

19. The method of claim 17 or claim 18, wherein the infection is caused by an enveloped virus.

20. The method according to any one of claims 17-19, wherein the infection is caused by a virus interacting with a cell expressing the ACE2 polypeptide.

21. The method according to any one of claims 17-20, wherein the viral infection is a coronavirus infection.

22. A method for inhibiting the interaction between a virus and cells expressing the ACE2 peptide, said method comprising, consisting of, or substantially consisting of: The virus is brought into contact with a compound according to any one of claims 1-14 or a pharmaceutically acceptable salt, solvate or prodrug thereof, or a compound of formula VII or a pharmaceutically acceptable salt, solvate or prodrug thereof.

23. The method of claim 22, wherein the interaction is one or both of virus binding to a cell and virus entry into a cell.

24. The method of claim 22 or claim 23, wherein the cells are lung cells (e.g., alveolar cells), intestinal cells, endothelial cells, epithelial cells (e.g., nasal or nasopharyngeal epithelial cells), kidney cells (e.g., brush border cells of proximal renal tubular epithelial cells), or arterial smooth muscle cells.

25. The method according to any one of claims 22-24, wherein the cell is a respiratory cell.

26. The method according to any one of claims 22-25, wherein the virus is a coronavirus.

27. A method for treating an acute inflammatory condition in a subject, wherein the condition is associated with coronavirus infection, the method comprising, consisting of, or substantially consisting of the following: The subject is given a compound according to any one of claims 1-14 or a pharmaceutically acceptable salt, solvate or prodrug thereof, or a compound of formula VII or a pharmaceutically acceptable salt, solvate or prodrug thereof.

28. A method for treating cytokine release syndrome (CRS) or cytokine storm in a subject, wherein the CRS or cytokine storm is associated with coronavirus infection, the method comprising, consisting of, or substantially consisting of the following: The subject is given a compound according to any one of claims 1-14 or a pharmaceutically acceptable salt, solvate or prodrug thereof, or a compound of formula VII or a pharmaceutically acceptable salt, solvate or prodrug thereof.

29. A method for treating SARS in a subject, wherein the SARS is associated with coronavirus infection, the method comprising, consisting of, or substantially consisting of: The subject is given a compound according to any one of claims 1-14 or a pharmaceutically acceptable salt, solvate or prodrug thereof, or a compound of formula VII or a pharmaceutically acceptable salt, solvate or prodrug thereof.

30. The method according to any one of claims 21 and 26-29, wherein the coronavirus is a beta-coronavirus.

31. The method of claim 30, wherein the beta coronavirus is selected from SARS-CoV and SARS-CoV-2.

32. The method of claim 31, wherein the beta coronavirus is SARS-CoV-2.

33. The method according to any one of claims 17-32, the method further comprising administering a second compound of formula I, II, III, IV, V, VI or VII, or a pharmaceutically acceptable salt, solvate or prodrug thereof.

34. Use of a compound according to any one of claims 1-14 or a pharmaceutically acceptable salt, solvate or prodrug thereof, or a compound of formula VII or a pharmaceutically acceptable salt, solvate or prodrug thereof, in the preparation of a medicament for treating or inhibiting the development of a viral infection in a subject.

35. Use of a compound according to any one of claims 1-14 or a pharmaceutically acceptable salt, solvate or prodrug thereof, or a compound of formula VII or a pharmaceutically acceptable salt, solvate or prodrug thereof, in the preparation of a medicament for treating an acute inflammatory condition in a subject, wherein the condition is associated with coronavirus infection.

36. Use of a compound according to any one of claims 1-14 or a pharmaceutically acceptable salt, solvate or prodrug thereof, or a compound of formula VII or a pharmaceutically acceptable salt, solvate or prodrug thereof, in the preparation of a medicament for treating a subject with CRS or cytokine storm, wherein the CRS or cytokine storm is associated with coronavirus infection.

37. Use in the preparation of a medicament of a compound according to any one of claims 1-14 or a pharmaceutically acceptable salt, solvate or prodrug thereof, or a compound of formula VII or a pharmaceutically acceptable salt, solvate or prodrug thereof, for treating SARS in a subject, wherein the SARS is associated with coronavirus infection.

38. The compound according to any one of claims 1-14, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, or the compound of formula VII, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, for the treatment or inhibition of the development of a viral infection in a subject.

39. The compound of any one of claims 1-14 or a pharmaceutically acceptable salt, solvate or prodrug thereof, or the compound of formula VII or a pharmaceutically acceptable salt, solvate or prodrug thereof, for treating an acute inflammatory condition in a subject, said condition being associated with coronavirus infection.

40. The compound of any one of claims 1-14 or a pharmaceutically acceptable salt, solvate or prodrug thereof, or the compound of formula VII or a pharmaceutically acceptable salt, solvate or prodrug thereof, for treating a subject with CRS or cytokine storm, wherein the CRS or cytokine storm is associated with coronavirus infection.

41. A compound according to any one of claims 1-14, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, or a compound of formula VII, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, for treating SARS in a subject, wherein the SARS is associated with coronavirus infection.

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