ROCK2 inhibitor for the treatment of viral infections

Abstract

The present disclosure provides a composition and method comprising a selective inhibitor of Rho-associated coiled-coil kinase 2 (ROCK2) for use in the treatment of viral infections, particularly in the treatment of coronavirus infections, such as SARS-CoV-2 infections, and in the treatment of sequelae resulting from viral infections, including sequelae resulting from coronavirus infections. The present invention provides, for example, a method for treating viral infection in a subject in need of treatment of viral infection, the method comprising administering to the subject a therapeutically effective amount of a ROCK2 inhibitor.

Description

Technical Field

[0001] Field The present disclosure relates to methods and compositions for the treatment of viral infections using Rho-associated coiled-coil kinase (ROCK) inhibitors, particularly inhibitors of Rho-associated coiled-coil kinase 2 (ROCK2).

Background Art

[0002] Background Viral infections are common in animals and humans. For example, the COVID-19 pandemic caused by the SARS-CoV-2 virus has threatened public health worldwide. Surprisingly, this pandemic has resulted in the infection of over 570 million people worldwide and the death of over 6 million people by mid-2022.

[0003] Coronaviruses are a large family of enveloped viruses with a positive - sense single - stranded RNA genome and belong to the family Coronaviridae and the order Nidovirales. Coronavirus infections mainly concentrate on the upper respiratory and gastrointestinal tracts, but in more severe infections, the lower respiratory tract can also be involved. Depending on the specific virus and host cell type, the symptoms and pathological damage caused by coronavirus infections can vary widely. Some coronaviruses, including HCoV - NL63, HCoV - 229E, and HCoV - OC43, continuously circulate in the human population and cause mild symptoms similar to the common cold. Other coronaviruses, including Severe Acute Respiratory Syndrome Coronavirus (SARS - CoV - 1), Middle East Respiratory Syndrome Coronavirus (MERS - CoV), and SARS - CoV - 2, can cause severe respiratory diseases with high morbidity and mortality. MERS - CoV was first reported in Saudi Arabia in 2012 and spread to several other countries. SARS - CoV - 1 was first recognized in China in 2002 and caused a global pandemic in 2002 and 2003. Other human coronaviruses include 229E (alpha - coronavirus), NL63 (alpha - coronavirus), OC43 (beta - coronavirus), and HKU1 (beta - coronavirus).

[0004] Persistent and diverse post - viral symptoms have been described in survivors of coronavirus infections, including survivors of Covid - 19, and even in those with a mild initial disease course. Currently, there is no curative treatment for post - viral syndrome, and therapies are directed at symptom relief and coping strategies. Furthermore, the economic impact of post - viral syndrome, including loss of productivity and employment, and increased need for disability benefits and financial support, can be substantial. There is a substantial and unmet need for therapies to treat coronavirus infections and the sequelae resulting from coronavirus infections.

[0005] Rho-associated coiled-coil kinase (ROCK) is a serine / threonine kinase derived from the AGC (PKA, PKG, and PKC) kinase family and includes two isoforms, ROCK1 and ROCK2. The two isoforms are differentially expressed and regulated in specific tissues. For example, ROCK1 is ubiquitously expressed at relatively high levels, while ROCK2 is preferentially expressed in certain tissues including the heart, brain, and skeletal muscle. ROCK is a target of the small GTPase Rho and is involved in diverse cellular activities achieved by phosphorylating downstream effector proteins (such as MLC, LIMK, ERM, MARCKS, CRMP-2, etc.). Studies have shown that various diseases (such as pulmonary fibrosis, cardiovascular diseases, neurological diseases, and cancer, etc.) are associated with the pathways mediated by ROCK. Therefore, ROCK is considered an important target in the development of new drugs.

[0006] The present disclosure relates to the use of ROCK2 inhibitors for the treatment of viral infections, including their previously unrecognized and surprisingly potent antiviral effects and coronavirus infections such as SARS-CoV-1 infection, SARS-CoV-2 infection, or MERS-CoV infection. Furthermore, ROCK2 inhibition treats many of the secondary conditions that can result from viral infection, including inflammation, fibrosis, and cytokine storms. As a result of treating such secondary conditions, ROCK2 inhibition may also be useful for treating or preventing long COVID. SUMMARY OF THE INVENTION MEANS FOR SOLVING THE PROBLEM

[0007] Overview In one aspect, the present disclosure provides an antiviral composition comprising a Rho-associated coiled-coil kinase 2 (ROCK2) inhibitor, such as a ROCK2 inhibitor disclosed herein. In one aspect, the present disclosure provides a method of treating a subject by administering to the subject a therapeutically effective amount of a ROCK2 inhibitor, such as a ROCK2 inhibitor disclosed herein, or a composition comprising a ROCK2 inhibitor. Such methods and compositions described herein may be useful for treating individuals affected by or suspected of having a viral infection. In embodiments, the viral infection is a coronavirus infection. In embodiments, the viral infection is a SARS-CoV-1 infection, a SARS-CoV-2 infection, or a MERS-CoV infection. In some embodiments, the viral infection is caused by SARS-CoV-2. In some embodiments, the viral infection is caused by the delta or omicron variant of SARS-CoV-2. In embodiments, the methods and compositions described herein are useful for treating and preventing sequelae resulting from viral infection, including sequelae resulting from coronavirus infection. In embodiments, the methods and compositions described herein are useful for treating and preventing sequelae resulting from SARS-CoV-1 infection, SARS-CoV-2 infection, or MERS-CoV infection. In some embodiments, the sequelae resulting from viral infection are caused by SARS-CoV-2. In some embodiments, the sequelae resulting from viral infection are caused by the delta or omicron variant of SARS-CoV-2. In embodiments, the sequelae include one or more of the group consisting of fatigue, dyspnea (shortness of breath), cough, arthralgia (joint pain), myalgia, headache, chest pain, fever, palpitations, myocarditis, ventricular dysfunction, stroke, pulmonary function abnormalities, pulmonary fibrosis, renal dysfunction, rash, alopecia, olfactory and / or gustatory dysfunction, sleep regulation abnormalities, cognitive function disorder changes, memory function disorder, depression, anxiety, mood swings, and combinations thereof. In embodiments, the sequelae include fibrosis. In some embodiments, the fibrosis is pulmonary fibrosis. As a result of treating one or more sequelae resulting from SARS-CoV-2 infection, a ROCK2 selective inhibitor may be useful for treating or preventing long COVID.

[0008] In one aspect, the present disclosure provides a method of treating viral infection in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a ROCK2 inhibitor, such as a ROCK2 selective inhibitor of the present disclosure. In some embodiments, the ROCK2 inhibitor is a ribonucleic acid (RNA). In some embodiments, the ROCK2 inhibitor is an antisense RNA against ROCK2 transcription. Optionally, the RNA is a small interfering RNA (siRNA) or a microRNA (miRNA). In some embodiments, the ROCK2 inhibitor is a compound having a structure of Formulas I-IV as defined herein, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotope-labeled compound thereof, particularly, (2-(3-(4-((1H-indazol-5-yl)amino)quinazolin-2-yl)phenoxy)-N-isopropylacetamide). In some embodiments, the ROCK2 inhibitor is Compound 1 as defined herein, or has a structure of a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotope-labeled compound thereof. In some embodiments, the ROCK2 inhibitor is belumosudil. In some embodiments, the subject is a human subject. In some embodiments, the subject is a veterinary subject.

[0009] In some embodiments, the method comprises administering to the subject a therapeutically effective amount of at least one other therapeutic agent. The at least one other therapeutic agent may be another antiviral agent, a corticosteroid, an anti-inflammatory signaling modulator, a β2-adrenergic receptor agonist bronchodilator, an anticholinergic agent, a mucolytic agent, hypertonic saline, or a combination thereof.

[0010] In one aspect, the present disclosure provides a method for treating viral infection in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a ROCK2 inhibitor, such as a ROCK2 selective inhibitor of the present disclosure, and a therapeutically effective amount of at least one other antiviral agent. In some embodiments, the ROCK2 inhibitor is a compound having a structure of Formulae I-IV as defined herein, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotopically labeled compound thereof, particularly (2-(3-(4-((1H-indazol-5-yl)amino)quinazolin-2-yl)phenoxy)-N-isopropylacetamide). In some embodiments, the ROCK2 inhibitor has the structure of Compound 1 as defined herein, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotopically labeled compound thereof. In some embodiments, the ROCK2 inhibitor is belumosudil. The at least one other antiviral agent may be a nucleoside or nucleotide analog, or a pharmaceutically acceptable salt or prodrug thereof. In some embodiments, the subject is a human subject. In some embodiments, the subject is a veterinary subject.

Brief Description of the Drawings

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Mode for Carrying Out the Invention

[0031] Detailed Description The compounds, compositions, and methods described herein are for use in the treatment of viral infections, particularly coronavirus infections, such as SARS-CoV-1 infection, SARS-CoV-2 infection (including all variants such as Delta and Omicron), or MERS-CoV infection, and in the treatment and prevention of sequelae resulting from viral infections, including sequelae resulting from coronavirus infections, and provide inhibitors of Rho-associated coiled-coil kinase 2 (ROCK2), including selective inhibitors of ROCK2. ROCK2 Inhibitor

[0032] Compounds for use in the methods and compositions disclosed herein are ROCK inhibitors, particularly ROCK2-selective inhibitors. The compounds provide excellent inhibitory activity against ROCK (preferably ROCK2), good selectivity (higher selectivity for ROCK2 compared to ROCK1), good physicochemical properties (e.g., solubility, physical and / or chemical stability), improved pharmacokinetic properties (e.g., improved bioavailability, appropriate half-life and duration of action), improved safety (low toxicity and / or fewer side effects, wide therapeutic window), and the like.

[0033] As provided herein, ROCK2 inhibitors have a surprisingly potent antiviral effect that has not been previously recognized. Without being bound by theory, for example, ROCK2 inhibitors such as antisense RNA, siRNA, or miRNA against ROCK2 transcription or compounds having the structures of Formulas I - IV defined herein can (1) the pathways used by the virus to enter the cell; (2) the cytoskeleton used by the virus as a path for migration and spread; (3) the pathways used by the virus to upregulate the cell's energy metabolism; and (4) one or more of the pathways used by the virus to spread to other cells. Inhibitors of ROCK2 interfere with viral interactions with cytoskeletal actin filaments, microtubules, and / or intermediate filaments, which are highly involved in the life cycle and pathological damage caused by viruses, particularly coronaviruses. Furthermore, ROCK2 inhibitors have a potent effect against many of the secondary conditions caused by viral infection, including inflammation, fibrosis, and cytokine and bradykinin storms. Thus, ROCK inhibitors may be useful for treating or preventing long COVID. Long COVID (also called post-COVID) is the appearance of new symptoms or the persistence of initial symptoms for 30 days after an initial COVID-19 infection. Long COVID can lead to post-COVID fibrosis, which can be seen on lung imaging by CT scan.

[0034] According to one aspect, the present disclosure provides a method of treating a subject by administering to the subject a therapeutically effective amount of a ROCK2 inhibitor, such as a ROCK2 selective inhibitor of the present disclosure, or a composition comprising a ROCK2 inhibitor. In some embodiments, the ROCK2 inhibitor is a ribonucleic acid (RNA). In some embodiments, the ROCK2 inhibitor is an antisense RNA against ROCK2 transcription. Optionally, the RNA is a small interfering RNA (siRNA) or a microRNA (miRNA). In some embodiments, the ROCK2 inhibitor has the structure of Formula I:

Chemical formula

[0035] In some embodiments, the ROCK2 inhibitor has the structure of formula II:

Chemical formula

[0036] In some embodiments, the ROCK2 inhibitor is a compound having the structure of Formula III:

Chemical formula

[0037] In some embodiments, the ROCK2 inhibitor has the structure of formula IV:

Chemical formula

[0038] In some embodiments, the ROCK2 inhibitor is a compound having the chemical structure of Compound 1 (2-(3-(4-((1H-indazol-5-yl)amino)quinazolin-2-yl)phenoxy)-N-isopropylacetamide):

Chemical formula

[0039] Compound 1 is a selective inhibitor of Rho-associated coiled-coil kinase 2 (ROCK2) in human cells. Virus infection can be treated using Compound 1 or a composition comprising Compound 1.

[0040] The compounds of Formulas I-IV, particularly, (2-(3-(4-((1H-indazol-5-yl)amino)quinazolin-2-yl)phenoxy)-N-isopropylacetamide), can be prepared according to the method disclosed in WO2014 / 055996, the entire content of which is incorporated herein by reference.

[0041] As used herein, the term “aliphatic” refers to straight-chain or branched alkyl, alkenyl, or alkynyl. It is understood that embodiments of alkenyl or alkynyl require at least 2 carbon atoms in the aliphatic chain. Aliphatic groups typically contain 1 (or 2) to 12 carbon atoms, such as 1 (or 2) to 4 carbon atoms.

[0042] As used herein, the term “alkylene” refers to a saturated divalent hydrocarbyl, preferably a saturated divalent hydrocarbyl having 1, 2, 3, 4, 5, or 6 carbon atoms, such as methylene (-CH2-), ethylene (-CH2CH2-), propylene, or butylene.

[0043] As used herein, the term “alkyl” is defined as a straight-chain or branched saturated aliphatic hydrocarbon. In some embodiments, alkyl has 1 to 12 carbon atoms, such as 1 to 6 carbon atoms. For example, as used herein, the term “C 1~6"Alkyl" refers to a straight-chain or branched group having 1 to 6 carbon atoms (such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, or n-hexyl), optionally substituted with one or more (e.g., 1 to 3) suitable substituents such as halogen (in this case, the group may be referred to as "haloalkyl") (e.g., CH2F, CHF2, CF3, CCl3, C2F5, C2Cl5, CH2CF3, CH2Cl, or CH2CH2CF3). The term "C 1~4 "Alkyl" refers to a straight-chain or branched aliphatic hydrocarbon chain having 1 to 4 carbon atoms (i.e., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl).

[0044] As used herein, the term "alkenyl" refers to a straight-chain or branched monovalent hydrocarbyl having a double bond and 2 to 6 carbon atoms ("C 2~6 alkenyl"). Alkenyls are, for example, vinyl, 1-propenyl, 2-propenyl, 2-butenyl, 3-butenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 2-methyl-2-propenyl, and 4-methyl-3-pentenyl. When the compounds of the present disclosure contain an alkylene group, the compounds may exist as a pure E (entgegen) form, a pure Z (zusammen) form, or any mixture thereof.

[0045] As used herein, the term "alkynyl" refers to a monovalent hydrocarbyl containing one or more triple bonds and preferably having 2, 3, 4, 5, or 6 carbon atoms, such as ethynyl or propynyl.

[0046] The term "cycloalkyl or cycloalkenyl" refers to a non-aromatic, monocyclic or fused or bridged bicyclic carbocyclic system. For example, the cycloalkyl or cycloalkenyl used herein may be a non-aromatic C3-C10 monocyclic or fused or bridged C8-C12 bicyclic carbocyclic system optionally substituted with one or more (e.g., 1 to 3) suitable substituents (e.g., bicyclo[1.1.1]pentyl, bicyclo[2.2.1]heptyl, bicyclo[3.2.1]octyl or bicyclo[5.2.0]nonyl, or decahydronaphthalene, etc.). The cycloalkenyl ring has one or more unsaturated units. Preferred cycloalkyl or cycloalkenyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl, norbornyl, adamantyl and decalinyl.

[0047] As used herein, the terms "cyclic hydrocarbylene", "cyclic hydrocarbyl" and "hydrocarbon ring" refer to saturated (i.e., "cycloalkylene" and "cycloalkyl") or unsaturated (i.e., having one or more double and / or triple bonds in the ring), monocyclic or polycyclic hydrocarbon rings having, for example, 3 to 10 (preferably 3 to 8, more preferably 3 to 6) ring carbon atoms, including, but not limited to, cyclopropyl(ene)(ring), cyclobutyl(ene)(ring), cyclopentyl(ene)(ring), cyclohexyl(ene)(ring), cycloheptyl(ene)(ring), cyclooctyl(ene)(ring), cyclononyl(ene)(ring), cyclohexenyl(ene)(ring), etc.

[0048] As used herein, the terms “heterocyclyl,” “heterocyclylene,” and “heterocycle” refer to a saturated (i.e., heterocycloalkyl) or partially unsaturated (i.e., having one or more double and / or triple bonds in the ring) cyclic group having at least one ring atom being a heteroatom selected from the group consisting of N, O, and S and the remaining ring atoms being C, for example, having 3 to 10 (preferably 3 to 8, more preferably 3 to 6) ring atoms. For example, “3- to 10-membered heterocyclyl(ene)” or “3- to 10-membered heterocycle” refers to a saturated or partially unsaturated heterocyclyl(ene) or heterocycle having 2 to 9 (e.g., 2, 3, 4, 5, 6, 7, 8, or 9) ring carbon atoms and one or more (e.g., 1, 2, 3, or 4) heteroatoms independently selected from the group consisting of N, O, and S. Examples of heterocyclylene, heterocyclyl, and heterocycle include, but are not limited to, oxiranyl(ene), aziridinyl(ene), azetidinyl(ene), oxetanyl(ene), tetrahydrofuranyl(ene), dioxolinyl(ene), pyrrolidinyl(ene), pyrrolidonyl(ene), imidazolidinyl(ene), pyrazolidinyl(ene), pyrrolinyl(ene), tetrahydropyranyl(ene), piperidinyl(ene), morpholinyl(ene), dithianyl(ene), thiomorpholinyl(ene), piperazinyl(ene), or trithianyl(ene). The groups also include bicyclic systems including spiro, fused, or bridged systems (e.g., 8-azaspiro[4.5]decane, 3,9-diazaspiro[5.5]undecane, 2-azabicyclo[2.2.2]octane, etc.). Heterocyclylene, heterocyclyl, and heterocycle may optionally be substituted with one or more (e.g., 1, 2, 3, or 4) suitable substituents.

[0049] As used herein, the term "aryl" refers to a monocyclic or bicyclic carbocyclic aromatic ring system having a conjugated π - electron system. The aryl used herein includes (C6 - C12)-aryl-. For example, the aryl used herein may be a C6 - C10 monocyclic or a C8 - C12 bicyclic carbocyclic aromatic ring system. In some embodiments, the aryl used herein may be (C6 - C10)-aryl-. Phenyl (or Ph) is an example of a monocyclic aromatic ring system. The bicyclic aromatic ring systems include 10 systems where both rings are aromatic, for example, naphthyl, and 10 systems where only one of the two rings is aromatic, for example, tetralin. The aryl(ene) or aromatic ring may be optionally substituted with one or more (such as 1 - 3) suitable substituents (for example, halogen, - OH, - CN, - NO2, and C 1~6 alkyl, etc.).

[0050] As used herein, the terms "heteroaryl(ene)" and "heteroaromatic ring" refer to monocyclic, bicyclic or tricyclic aromatic ring systems having 5, 6, 8, 9, 10, 11, 12, 13 or 14 ring atoms, particularly having 1 or 2 or 3 or 4 or 5 or 6 or 9 or 10 carbon atoms and containing at least one heteroatom (such as O, N, or S), which may be the same or different. Further, in any case, it may be benzo - fused. In particular, "heteroaryl(ene)" or "heteroaromatic ring" is selected from the group consisting of thienyl(ene), furyl(ene), pyrrolyl(ene), oxazolyl(ene), thiazolyl(ene), imidazolyl(ene), pyrazolyl(ene), isoxazolyl(ene), isothiazolyl(ene), oxadiazolyl(ene), triazolyl(ene), thiadiazolyl(ene), etc., and their benzo - derivatives; or pyridinyl(ene), pyridazinyl(ene), pyrimidinyl(ene), pyrazinyl(ene), triazinyl(ene), etc., and their benzo - derivatives.

[0051] As used herein, the term "aralkyl" preferably means alkyl substituted with aryl or heteroaryl, where aryl, heteroaryl and alkyl are as defined herein. Usually, an aryl group may have 6 to 14 carbon atoms, a heteroaryl group may have 5 to 14 ring atoms, and an alkyl group may have 1 to 6 carbon atoms. Exemplary aralkyl groups include, but are not limited to, benzyl, phenylethyl, phenylpropyl, and phenylbutyl.

[0052] As used herein, the term "halo" or "halogen" is defined to include F, Cl, Br, or I.

[0053] As used herein, the term "nitrogen-containing heterocycle" refers to a saturated or unsaturated monocyclic or bicyclic group having 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13 carbon atoms and at least 1 nitrogen atom in the ring, which may further contain one or more (e.g., 1, 2, 3 or 4) ring members selected from the group consisting of N, O, C=O, S, S=O and S(=O)2 as required. The nitrogen-containing heterocycle is attached to the rest of the molecule via the nitrogen atom and any other ring atoms in the nitrogen-containing heterocycle. The nitrogen-containing heterocycle may be optionally benzo-fused, preferably attached to the rest of the molecule via the nitrogen atom in the nitrogen-containing heterocycle and any carbon atom in the fused benzene ring.

[0054] As used herein, the term "glycoside" refers to any material having a chemical structure containing a glycosidic bond between a carbohydrate (sugar) molecule and another carbohydrate or non-carbohydrate (non-sugar) moiety. A glycosidic bond or glycosidic linkage is a type of covalent bond that connects a carbohydrate (sugar) molecule to another molecule, for example, via its hemiacetal or hemiketal group. The other molecule may or may not be a carbohydrate. The sugar moiety is generally known as the glycone moiety of the glycoside. The glycone may consist of a single sugar group (monosaccharide) or several sugar groups (oligosaccharide).

[0055] As used herein, the term "substituent" refers to a group that replaces a second atom or group, such as a hydrogen atom, on any molecule, compound, or moiety. Suitable substituents include, without limitation, halo, hydroxy, mercapto, oxo, nitro, haloalkyl, alkyl, alkaryl, aryl, aralkyl, alkoxy, thioalkoxy, aryloxy, amino, alkoxycarbonyl, amide, carboxy, alkanesulfonyl, alkylcarbonyl, and cyano groups.

[0056] As used herein, the term "derivative" refers to a compound that retains the biological activity of the parent compound (from which the derivative is derived) or is a prodrug for the parent compound. Derivatives may include esters, amides, and ethers of the parent compound, obtained by chemically modifying moieties of the parent compound. For example, a derivative may be a compound in which a hydrogen atom or a particular atomic group is replaced by another atom or atomic group.

[0057] The term "substituted" means that one or more (e.g., 1, 2, 3, or 4) hydrogens on the designated atom are replaced with a selection from the indicated groups, provided that the substitution does not exceed the normal valence of the designated atom under existing circumstances and results in a stable compound. Combinations of substituents and / or variables are permitted only if such combinations result in a stable compound.

[0058] When a substituent is "optionally substituted", the substituent may (1) be unsubstituted or (2) be substituted. When it is described that a carbon of a substituent is optionally substituted with one or more of a list of substituents, one or more (to the extent present) of the hydrogens on the carbon may be replaced separately and / or together with an independently selected substituent as required. When a nitrogen of a substituent is optionally substituted with one or more of a list of substituents, one or more (to the extent present) of the hydrogens on the nitrogen may each be replaced with an independently selected substituent as required.

[0059] When a substituent is described as being "independently selected" from a group, each substituent is selected independently of the others. Each substituent may thus be the same as or different from the other substituents.

[0060] As used herein, the term "one or more" means one or more than one (e.g., 2, 3, 4, 5, or 10), as would be reasonable.

[0061] As used herein, unless otherwise specified, the point of attachment of a substituent may be at any suitable position of the substituent.

[0062] When a bond to a substituent is shown to cross a bond connecting two atoms in a ring, such a substituent can be attached to any of the ring-forming atoms in that ring that are substitutable.

[0063] Compounds for use in the methods provided herein include pharmaceutically acceptable, isotopically labeled compounds that are identical to those of Formulas I-IV, except that one or more atoms have been replaced with atoms having an atomic mass or mass number different from the atomic mass or mass number that is predominant in nature. Examples of suitable isotopes for inclusion in the compounds include, but are not limited to, 2 H, 3 hydrogen such as 11 C,13 C and 14 carbons such as C; 36 chlorines such as Cl; 18 fluorines such as F; 123 I and 125 iodines such as I; 13 N and 15 nitrogens such as N; 15 O, 17 O and 18 oxygens such as O; 32 phosphoruses such as P; and 35 sulfur isotopes such as S are included. Certain isotope-labeled compounds of the present disclosure, for example, those incorporating radioisotopes, are useful in drug and / or substrate tissue distribution studies (e.g., assays). Tritium, which is a radioisotope, that is, 3 H, and carbon-14, that is, 14 C are particularly useful for this purpose considering the ease of their incorporation and the easy means of detection. 11 C, 18 F, 15 O and 13 replacement with positron-emitting isotopes such as N may be useful in positron emission tomography (PET) tests for examining substrate receptor occupancy. The isotope-labeled compounds of the present disclosure can generally be prepared by using appropriate isotope-labeled reagents instead of the previously used unlabeled reagents, by processes similar to those described in the attached schemes and / or examples and preparations. Pharmaceutically acceptable solvates according to the present disclosure include those in which the crystallization solvent may be isotope-substituted, for example, D2O, acetone-d6, or DMSO-d6.

[0064] The term "stereoisomer" refers to isomers having at least one asymmetric center. Compounds having one or more (e.g., 1, 2, 3, or 4) asymmetric centers can give rise to racemic mixtures, single enantiomers, diastereomeric mixtures, and individual diastereomers. Certain individual molecules may exist as geometric isomers (cis / trans). Similarly, the compounds provided herein may exist as mixtures of two or more structurally different forms in rapid equilibrium (generally referred to as tautomers). Typical examples of tautomers include keto-enol tautomers, phenol-keto tautomers, nitroso-oxime tautomers, imine-enamine tautomers, and the like. It should be understood that the use of all such isomers and mixtures thereof in any proportion (such as 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, and 99%) is encompassed within the scope of the present disclosure.

[0065] The present disclosure includes the use of any possible crystalline forms or polymorphs of the compounds of the present disclosure as a single polymorph or as a mixture of more than one polymorph in any ratio.

[0066] It should also be understood that certain compounds provided herein can be used for treatment in free form or, where appropriate, in the form of pharmaceutically acceptable derivatives. In the present disclosure, pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable salts, esters, solvates, N-oxides, metabolites, or prodrugs that can directly or indirectly provide the compounds of the present disclosure or their metabolites or residues after administration to a patient in need thereof. Accordingly, the compounds provided herein include various derivative forms of the above compounds.

[0067] The pharmaceutically acceptable salts of the compounds disclosed herein include acid addition salts or base addition salts. Suitable acid addition salts are formed from acids that form pharmaceutically acceptable salts. Specific non-limiting examples include acetate, adipate, aspartate, benzoate, besylate, bicarbonate / carbonate, bisulfate / sulfate, borate, camphorsulfonate, citrate, cyclamate, edisylate, esylate, formate, fumarate, gluceptate, gluconate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride / chloride, hydrobromide / bromide, hydroiodide / iodide, isethionate, lactate, malate, maleate, malonate, mesylate, methylsulfate, naphthylate, 2-napsylate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate / hydrogen phosphate / dihydrogen phosphate, pyroglutamate, saccharate, stearate, succinate, tannate, tartrate, tosylate, trifluoroacetate, and xinofoate salts. Suitable base addition salts are formed from bases that form pharmaceutically acceptable salts. Specific non-limiting examples include aluminum salts, arginine salts, benzathine salts, calcium salts, choline salts, diethylamine salts, diolamine salts, glycine salts, lysine salts, magnesium salts, meglumine salts, olamine salts, potassium salts, sodium salts, tromethamine salts, and zinc salts.

[0068] For a general overview of suitable pharmaceutically acceptable salts, see "Handbook of Pharmaceutical Salts: Properties, Selection, and Use", Stahl and Wermuth (Wiley-VCH, 2002).

[0069] As used herein, the term "ester" refers to those derived from compounds of various formulas provided herein, including physiologically hydrolysable esters (which can be hydrolyzed under physiological conditions to release the compounds of the present disclosure in the form of the free acid or alcohol).

[0070] Compounds for use in the methods of the present disclosure may exist as solvates (preferably, hydrates), and the compounds contain a polar solvent, particularly, for example, water, methanol, or ethanol, as a structural element of the crystal lattice of the compound. The amount of the polar solvent, particularly water, may be present in a stoichiometric or non-stoichiometric ratio.

[0071] As will be appreciated by those skilled in the art, not all nitrogen-containing heterocycles can form N-oxides because nitrogen requires lone pairs of electrons available for oxidation to oxides; those skilled in the art will recognize nitrogen-containing heterocycles that can form N-oxides. Those skilled in the art will also recognize that tertiary amines can form N-oxides. Synthetic methods for the preparation of N-oxides of heterocycles and tertiary amines are well known to those skilled in the art and include the oxidation of heterocycles and tertiary amines with peroxyacids such as peracetic acid and m-chloroperbenzoic acid (mCPBA), hydrogen peroxide, alkyl hydroperoxides such as tert-butyl hydroperoxide, sodium perborate, and dioxiranes such as dimethyldioxirane. These methods for the preparation of N-oxides are widely described and reviewed in the literature; see, for example, T. L. Gilchrist, Comprehensive Organic Synthesis, vol. 7, pp 748-750; A. R. Katritzky and A. J. Boulton, Eds., Academic Press; and G W. H. Cheeseman and E. S. G Werstiuk, Advances in Heterocyclic Chemistry, vol. 22, pp 390-392, A. R. Katritzky and A. J. Boulton, Eds., Academic Press.

[0072] The compounds described in this specification can be administered in the form of prodrugs, which, when administered in vivo or on the body to certain derivatives of compounds that may have little or no pharmacological activity per se, can be converted to compounds having the desired activity, for example, by hydrolytic cleavage. Generally, such prodrugs are functional derivatives of compounds that are readily convertible in vivo to compounds having the desired therapeutic activity. Further information regarding the use of prodrugs can be found in "Pro-drugs as Novel Delivery Systems, Vol. 14, ACS Symposium Series (T. Higuchi and V. Stella). For example, prodrugs can be produced by replacing appropriate functional groups present in the compounds of the present disclosure with moieties known to those skilled in the art as "pro-moieties", as described in, for example, "Design of Prodrugs", H. Bundgaard (Elsevier, 1985).

[0073] In some embodiments, the compounds of Formulas I-IV are selective inhibitors of Rho-associated coiled-coil kinase 2 (ROCK2) in human cells. For example, the compounds of Formulas I-IV as pharmaceutical compositions containing the compounds are used to treat (i.e., cure or reduce the severity of, etc.) viral infections, particularly coronavirus infections such as SARS-CoV-1, SARS-CoV-2, and MERS-CoV, and to treat or prevent sequelae resulting from viral infections, including coronavirus infections such as SARS-CoV-1, SARS-CoV-2, and MERS-CoV. In some embodiments, the viral infection is a SARS-CoV-1 infection. In some embodiments, the viral infection is a SARS-CoV-2 infection. In some embodiments, the viral infection is caused by the delta or omicron variant of SARS-CoV-2. In some embodiments, the viral infection is a MERS-CoV infection.

[0074] Methods for determining kinase inhibition are disclosed herein. For example, the kinase activity of an enzyme and the inhibitory ability of a test compound can be determined by measuring the enzyme-specific phosphorylation of a substrate. Commercially available assays and kits can be used. For example, kinase inhibition can be determined using the IMAP® assay (Molecular Devices). This assay method involves the use of a fluorescently tagged peptide substrate. Phosphorylation of the tagged peptide by the kinase of interest promotes the binding of the peptide to trivalent metal-based nanoparticles via specific high-affinity interactions between the phosphate group and the trivalent metal. Proximity to the nanoparticles results in an increase in fluorescence polarization. Inhibition of the kinase by a kinase inhibitor prevents phosphorylation of the substrate, thereby limiting the binding of the fluorescently tagged substrate to the nanoparticles. Such assays are compatible with the microplate assay format and may enable the simultaneous determination of the IC 50 values of multiple compounds.

[0075] In some embodiments, the ROCK2 inhibitor is not a compound disclosed in US2019 / 0276440, nor a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotopically labeled compound of a compound disclosed in US2019 / 0276440.

[0076] In some embodiments, the ROCK2 inhibitor is a compound of formula V: [Chemical formula] or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotopically labeled compound thereof (wherein X and Y are each independently selected from the group consisting of a direct bond, C(=O), O, S(=O) i and NR; R is H, C 1~6 alkyl, C 2~6 alkenyl, C 2~6 alkynyl, saturated or partially unsaturated C 3~10Cyclic hydrocarbyl, saturated or partially unsaturated 3- to 10-membered heterocyclyl, C 6~10 Aryl, 5- to 14-membered heteroaryl and C 6~12 Selected from the group consisting of aralkyl, and at most two ring members in the cyclic hydrocarbyl and heterocyclyl are C(=O); Ring A and ring B are each independently selected from the group consisting of saturated or partially unsaturated C 3~10 Hydrocarbon ring, saturated or partially unsaturated 3- to 10-membered heterocyclic ring, C 6~10 Aromatic ring and 5- to 14-membered heteroaromatic ring, and at most two ring members in the hydrocarbon ring and heterocyclic ring are C(=O), provided that when ring B is a heterocyclic ring containing a nitrogen atom, ring B is not bonded to X through the nitrogen atom; Ring C is selected from the group consisting of saturated or partially unsaturated C 3~10 Hydrocarbon ring, saturated or partially unsaturated 3- to 10-membered heterocyclic ring, C 6~10 Aromatic ring and 5- to 14-membered heteroaromatic ring, and at most two ring members in the hydrocarbon ring and heterocyclic ring are C(=O); Ring D is absent or is selected from the group consisting of saturated or partially unsaturated C 3~10 Hydrocarbon ring, saturated or partially unsaturated 3- to 10-membered heterocyclic ring, C 6~10 Aromatic ring and 5- to 14-membered heteroaromatic ring, and at most two ring members in the hydrocarbon ring and heterocyclic ring are C(=O); Ring E is

Chemical Structure

Chemical formula

[0077] In some embodiments, the ROCK2 inhibitor is a compound or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotopically labeled compound according to any one of Formulas VI-XIII: [Chemical formula] [Chemical formula] [Chemical formula] wherein ring A, ring B, ring D, R, R 1 、R 1a 、R 1b 、R 2 、R 3 、R 4 、R 7 、R 7’ 、R 8 、R 9 、R 10 、n and m are as defined in Formula V above) is not.

[0078] In some embodiments, the ROCK2 inhibitor is a compound of formula XIV or XV):

Chemical formula

Chemical formula

[0079] In some embodiments, the ROCK2 inhibitor is not a compound having the chemical structure of Compound 2:

Chemical formula

[0080] In some embodiments, the ROCK2 inhibitor is not (6-(4-((4-(1H-pyrazol-4-yl)phenyl)amino)pyrimidin-2-yl)-1-methyl-1H-indol-2-yl)(3,3-difluoroazetidin-1-yl)methanone or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotopically labeled compound thereof.

[0081] In some embodiments, the viral infection is not a coronavirus infection. In some embodiments, the viral infection is not an SARS-CoV-1 infection, an SARS-CoV-2 infection, or a MERS-CoV infection.

[0082] In some embodiments, when the viral infection is a coronavirus infection, the ROCK2 inhibitor is not a compound described in any one of Formulas V to XV. In some embodiments, when the viral infection is a coronavirus infection, the ROCK2 inhibitor is not a compound described in Formula V. In some embodiments, when the viral infection is a coronavirus infection, the ROCK2 inhibitor is not a compound described in Formula VI. In some embodiments, when the viral infection is a coronavirus infection, the ROCK2 inhibitor is not a compound described in Formula VII. In some embodiments, when the viral infection is a coronavirus infection, the ROCK2 inhibitor is not a compound described in Formula VIII. In some embodiments, when the viral infection is a coronavirus infection, the ROCK2 inhibitor is not a compound described in Formula IX. In some embodiments, when the viral infection is a coronavirus infection, the ROCK2 inhibitor is not a compound described in Formula X. In some embodiments, when the viral infection is a coronavirus infection, the ROCK2 inhibitor is not a compound described in Formula XI. In some embodiments, when the viral infection is a coronavirus infection, the ROCK2 inhibitor is not a compound described in Formula XII. In some embodiments, when the viral infection is a coronavirus infection, the ROCK2 inhibitor is not a compound described in Formula XIII. In some embodiments, when the viral infection is a coronavirus infection, the ROCK2 inhibitor is not a compound described in Formula XIV. In some embodiments, when the viral infection is a coronavirus infection, the ROCK2 inhibitor is not a compound described in Formula XV. In some embodiments, when the viral infection is a coronavirus infection, the ROCK2 inhibitor is not Compound 2.

[0083] In some embodiments, when the viral infection is SARS-CoV-1 infection, SARS-CoV-2 infection, or MERS-CoV infection, the ROCK2 inhibitor is not a compound described in any one of Formulas V to XV. In some embodiments, when the viral infection is SARS-CoV-1 infection, SARS-CoV-2 infection, or MERS-CoV infection, the ROCK2 inhibitor is not a compound described in Formula V. In some embodiments, when the viral infection is SARS-CoV-1 infection, SARS-CoV-2 infection, or MERS-CoV infection, the ROCK2 inhibitor is not a compound described in Formula VI. In some embodiments, when the viral infection is SARS-CoV-1 infection, SARS-CoV-2 infection, or MERS-CoV infection, the ROCK2 inhibitor is not a compound described in Formula VII. In some embodiments, when the viral infection is SARS-CoV-1 infection, SARS-CoV-2 infection, or MERS-CoV infection, the ROCK2 inhibitor is not a compound described in Formula VIII. In some embodiments, when the viral infection is SARS-CoV-1 infection, SARS-CoV-2 infection, or MERS-CoV infection, the ROCK2 inhibitor is not a compound described in Formula IX. In some embodiments, when the viral infection is SARS-CoV-1 infection, SARS-CoV-2 infection, or MERS-CoV infection, the ROCK2 inhibitor is not a compound described in Formula X. In some embodiments, when the viral infection is SARS-CoV-1 infection, SARS-CoV-2 infection, or MERS-CoV infection, the ROCK2 inhibitor is not a compound described in Formula XI. In some embodiments, when the viral infection is SARS-CoV-1 infection, SARS-CoV-2 infection, or MERS-CoV infection, the ROCK2 inhibitor is not a compound described in Formula XII. In some embodiments, when the viral infection is SARS-CoV-1 infection, SARS-CoV-2 infection, or MERS-CoV infection, the ROCK2 inhibitor is not a compound described in Formula XIII. In some embodiments, when the viral infection is SARS-CoV-1 infection, SARS-CoV-2 infection, or MERS-CoV infection, the ROCK2 inhibitor is not a compound described in Formula XIV.In some embodiments, when the viral infection is a SARS-CoV-1 infection, a SARS-CoV-2 infection, or a MERS-CoV infection, the ROCK2 inhibitor is not a compound as described in formula XV. In some embodiments, when the viral infection is a SARS-CoV-1 infection, a SARS-CoV-2 infection, or a MERS-CoV infection, the ROCK2 inhibitor is not compound 2.

[0084] In some embodiments, when the viral infection is a SARS-CoV-2 infection, the ROCK2 inhibitor is not a compound described in any one of Formulas V to XV. In some embodiments, when the viral infection is a SARS-CoV-2 infection, the ROCK2 inhibitor is not a compound described in Formula V. In some embodiments, when the viral infection is a SARS-CoV-2 infection, the ROCK2 inhibitor is not a compound described in Formula VI. In some embodiments, when the viral infection is a SARS-CoV-2 infection, the ROCK2 inhibitor is not a compound described in Formula VII. In some embodiments, when the viral infection is a SARS-CoV-2 infection, the ROCK2 inhibitor is not a compound described in Formula VIII. In some embodiments, when the viral infection is a SARS-CoV-2 infection, the ROCK2 inhibitor is not a compound described in Formula IX. In some embodiments, when the viral infection is a SARS-CoV-2 infection, the ROCK2 inhibitor is not a compound described in Formula X. In some embodiments, when the viral infection is a SARS-CoV-2 infection, the ROCK2 inhibitor is not a compound described in Formula XI. In some embodiments, when the viral infection is a SARS-CoV-2 infection, the ROCK2 inhibitor is not a compound described in Formula XII. In some embodiments, when the viral infection is a SARS-CoV-2 infection, the ROCK2 inhibitor is not a compound described in Formula XIII. In some embodiments, when the viral infection is a SARS-CoV-2 infection, the ROCK2 inhibitor is not a compound described in Formula XIV. In some embodiments, when the viral infection is a SARS-CoV-2 infection, the ROCK2 inhibitor is not a compound described in Formula XV. In some embodiments, when the viral infection is a SARS-CoV-2 infection, the ROCK2 inhibitor is not Compound 2. Pharmaceutical composition

[0085] In one aspect, the present disclosure provides a pharmaceutically acceptable composition for use in the treatment of viral diseases, comprising one or more therapeutically effective amounts of a ROCK2 inhibitor formulated with one or more pharmaceutically acceptable carriers, wherein the ROCK2 inhibitor is as disclosed herein. In some embodiments, the ROCK2 inhibitor is a ribonucleic acid (RNA). In some embodiments, the ROCK2 inhibitor is an antisense RNA against ROCK2 transcription. Optionally, the RNA is a small interfering RNA (siRNA) or a microRNA (miRNA). In some embodiments, the ROCK2 inhibitor is a compound of Formula I-IV as defined herein, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotopically labeled compound thereof, particularly, (2-(3-(4-((1H-indazol-5-yl)amino)quinazolin-2-yl)phenoxy)-N-isopropylacetamide). In some embodiments, the ROCK2 inhibitor has the structure of Compound 1 as defined herein, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotopically labeled compound thereof. In some embodiments, the ROCK2 inhibitor is belumosudil.

[0086] As detailed below, the pharmaceutical compositions of the present disclosure are adapted for: (1) oral administration, for example, a drinkable medicine (aqueous or non-aqueous solution or suspension), tablets, for example, buccal, sublingual, and those targeted for systemic absorption, bolus agents, powders, granules, pastes for application to the tongue; (2) parenteral administration, for example, sterile solutions or suspensions, or parenteral administration by, for example, subcutaneous, intramuscular, intravenous or epidural injection as a sustained release formulation; (3) topical application, for example, as a cream, ointment, or controlled release patch or spray applied to the skin; (4) intravaginal or rectal, for example, as a suppository, pessary, cream or foam; (5) sublingual; (6) intraocular; (7) transdermal; or (8) nasal, and can be specially formulated for administration in solid or liquid form.

[0087] As used herein, the phrase "pharmaceutically acceptable" refers to compounds, materials, compositions, and / or dosage forms that are suitable for use in contact with human and animal tissues within the scope of sound medical judgment, and that have a reasonable benefit / risk ratio, and are associated with no toxicity, irritation, allergic response, or other problems or complications.

[0088] As used herein, the phrase "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, manufacturing aid (e.g., lubricants, magnesium stearate, calcium stearate or zinc stearate, or stearic acid), solvent, or encapsulating material, involved in carrying or transporting a subject compound from one organ, or portion of the body, to another organ, or portion of the body. Each carrier should be compatible with the other ingredients of the formulation and should not be harmful to the patient. Some examples of materials that can function as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; (4) tragacanth powder; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol, and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) pH buffering solutions; (21) polyesters, polycarbonates, and / or polyanhydrides; and (22) other non-toxic compatible substances used in pharmaceutical formulations.

[0089] The ROCK2 inhibitors, including the ROCK2 selective inhibitors disclosed herein, can be formulated with conventional carriers and excipients that can be selected according to normal practice. Tablets may contain excipients, glidants, fillers, binders, etc. Aqueous formulations are prepared in a sterile form and may generally be isotonic if delivery other than oral administration is intended. All formulations may contain excipients such as those described in "Handbook of Pharmaceutical Excipients" (1986) if necessary. Excipients include ascorbic acid and other antioxidants, chelating agents such as EDTA, carbohydrates such as dextran, hydroxyalkyl cellulose, hydroxyalkyl methyl cellulose, stearic acid, and the like.

[0090] The ROCK2 inhibitors, including the ROCK2 selective inhibitors disclosed herein, can be administered alone, but may preferably be presented as pharmaceutical formulations. Formulations for both veterinary and human use according to the present disclosure include at least one ROCK2 inhibitor as provided above, together with one or more acceptable carriers therefor, and optionally other therapeutic components, particularly additional therapeutic components contemplated herein.

[0091] Formulations include those suitable for the routes of administration provided herein. The formulations can be conveniently presented in unit dosage form and can be prepared by any of the methods well known in the pharmaceutical industry. Techniques and formulations are generally found in Remington's Pharmaceutical Sciences (Mack Publishing Co., Easton, Pa.). Such methods include the step of bringing the ROCK2 inhibitor into association uniformly and closely with a carrier that constitutes one or more accessory components. Generally, the formulations are prepared by bringing the ROCK2 inhibitor into intimate association uniformly with a liquid carrier or a micronized solid carrier or both, and then, if necessary, shaping the product.

[0092] The formulations of the present disclosure suitable for oral administration can be presented as individual units such as capsules, cachets or tablets, each containing a predetermined amount of a ROCK2 inhibitor; as powders or granules; as solutions or suspensions in aqueous or non-aqueous liquids; or as water-in-oil or oil-in-water liquid emulsions. The ROCK2 inhibitor can also be administered as a bolus, a liniment or a paste.

[0093] Tablets can be prepared by compression or molding, optionally with one or more accessory ingredients. Compressed tablets can be prepared by compressing, in a suitable machine, a free-flowing form of the ROCK2 inhibitor, such as a powder or granules, mixed, optionally, with a binder, a lubricant, an inert diluent, a preservative, a surfactant or a dispersing agent. Molded tablets can be prepared by molding, in a suitable machine, a mixture of the powdered ROCK2 inhibitor moistened with an inert liquid diluent. The tablets may be optionally coated or grooved and formulated, optionally, to provide for a delayed or controlled release of the ROCK2 inhibitor therefrom.

[0094] For eye or other external tissue infections, such as those of the mouth and skin, the formulation is preferably applied as a topical solution, ointment or cream containing the ROCK2 inhibitor. The ROCK2 inhibitor may be present, for example, in an amount of 0.075 to 20% w / w (including ROCK2 inhibitors in the range of 0.1% to 20% in increments of 0.1% w / w such as 0.6% w / w, 0.7% w / w, etc.), preferably 0.2 to 15% w / w, most preferably 0.5 to 10% w / w. When formulated into an ointment, the ROCK2 inhibitor can be used with a paraffinic or water-miscible ointment base. Alternatively, the ROCK2 inhibitor can be formulated into a cream with a water-in-oil cream base.

[0095] Optionally, the aqueous phase of the cream base may contain, for example, at least 30% w / w of a polyhydric alcohol, i.e., propylene glycol, butane 1,3-diol, mannitol, sorbitol, glycerol, and polyethylene glycol (including PEG400) and mixtures thereof, etc., an alcohol having two or more hydroxyl groups. The topical formulation may desirably also contain a compound that promotes the absorption or penetration of the ROCK2 inhibitor through the skin or other affected areas. Examples of such skin penetration enhancers include dimethyl sulfoxide and related analogs.

[0096] The oil phase of the emulsion of the present disclosure may be composed of known components in a known manner. The phase may simply contain an emulsifier (otherwise known as an emulgent), but it desirably contains at least one emulsifier and a mixture of fat or oil, or both fat and oil. Preferably, a hydrophilic emulsifier is included together with a lipophilic emulsifier that acts as a stabilizer. It is also preferred to include both oil and fat. Collectively, the emulsifier, with or without a stabilizer, constitutes a so-called emulsifying wax, and the wax together with the oil and fat constitutes a so-called emulsifying ointment base that forms the oily dispersion phase of the cream formulation.

[0097] Emulsifiers and emulsion stabilizers suitable for use in the formulations of the present disclosure include Tween® 60, Span® 80, cetostearyl alcohol, benzyl alcohol, myristyl alcohol, glyceryl monostearate, and sodium lauryl sulfate. Further emulsifiers and emulsion stabilizers suitable for use in the formulations of the present disclosure include Tween® 80.

[0098] The selection of a suitable oil or fat for the formulation is based on achieving the desired properties. The cream should preferably be an oily-free, non-staining and washable product with a suitable consistency to avoid leakage from tubes or other containers. Straight or branched chain, mono- or dibasic alkyl esters such as diisoadipic acid esters, isocetyl stearate, propylene glycol diesters of coconut fatty acids, isopropyl myristate, decyl oleate, isopropyl palmitate, butyl stearate, 2-ethylhexyl palmitate or blends of branched chain esters known as Crodamol CAP can be used, with the last three being preferred esters. Depending on the required properties, these may be used alone or in combination. Alternatively, high melting point lipids such as white petrolatum and / or liquid paraffin or other mineral oils can be used.

[0099] The pharmaceutical preparations according to the present disclosure include one or more pharmaceutically acceptable carriers or excipients and, optionally, the combination according to the present disclosure together with other therapeutic agents. The pharmaceutical preparation containing a ROCK2 inhibitor may be in any form suitable for the intended method of administration. For example, when used for oral use, tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, syrups or elixirs can be prepared. The compositions intended for oral use can be prepared according to any method known in the art for the manufacture of pharmaceutical compositions, and such compositions may contain one or more agents including sweetening agents, flavoring agents, coloring agents and preservatives for providing a palatable preparation. Tablets containing a ROCK2 inhibitor admixed with non-toxic pharmaceutically acceptable excipients suitable for the manufacture of tablets are acceptable. These excipients may be, for example, inert diluents such as calcium carbonate or sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents such as corn starch, or alginic acid; binding agents such as starch, gelatin or acacia; and lubricating agents such as magnesium stearate, stearic acid or talc, etc. The tablets may not be coated or may be coated by known techniques including microencapsulation to delay disintegration and adsorption in the gastrointestinal tract, thereby providing a sustained action over a longer period. For example, time-delay materials such as glyceryl monostearate or glyceryl distearate may be used alone or in combination with waxes.

[0100] The preparations for oral use can also be presented as hard gelatin capsules in which the ROCK2 inhibitor is mixed with an inert solid diluent such as starch, mannitol, calcium phosphate or kaolin, or as soft gelatin capsules in which the ROCK2 inhibitor is mixed with a water or oil medium such as peanut oil, liquid paraffin or olive oil.

[0101] The aqueous suspensions of the present disclosure contain an active material admixed with excipients suitable for the manufacture of aqueous suspensions. Such excipients include suspending agents such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, tragacanth gum and acacia gum, and dispersing or wetting agents such as naturally occurring phosphatides (e.g., lecithin), condensation products of alkylene oxides with fatty acids (e.g., polyoxyethylene stearate), condensation products of ethylene oxide with long-chain fatty alcohols (e.g., heptadecaethyleneoxycetanol), condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides (e.g., polyoxyethylene sorbitan monooleate), and the like. The aqueous suspensions may also contain one or more preservatives such as ethyl or n-propyl p-hydroxy-benzoic acid, one or more colorants, one or more flavoring agents and one or more sweetening agents such as sucrose or saccharin. Further non-limiting examples of suspending agents include cyclodextrin and Captisol (= sulfobutyl ether beta-cyclodextrin; SEB-beta-CD).

[0102] The ROCK2 inhibitor can be formulated into an oil suspension by suspending it in a vegetable oil such as peanut oil, olive oil, sesame oil or coconut oil, or a mineral oil such as liquid paraffin. Oral suspensions may contain a thickening agent such as beeswax, solid paraffin or cetyl alcohol. Sweetening agents such as those described above and flavoring agents can be added to provide an orally palatable preparation. These compositions can be preserved by the addition of an antioxidant such as ascorbic acid.

[0103] The dispersible powders and granules of the present disclosure, which are suitable for the preparation of aqueous suspensions by the addition of water, provide ROCK2 inhibitors admixed with a dispersant or wetting agent, a suspending agent, and one or more preservatives. Suitable dispersants or wetting agents and suspending agents are exemplified by those disclosed above. Additional excipients, such as sweetening agents, flavoring agents, and coloring agents, may be present.

[0104] The pharmaceutical compositions of the present disclosure may also be in the form of oil-in-water emulsions. The oil phase may be a vegetable oil, such as olive oil or peanut oil, a mineral oil, such as liquid paraffin, or a mixture thereof. Suitable emulsifying agents include naturally occurring gums, such as acacia gum and tragacanth gum, naturally occurring phosphatides, such as soybean lecithin, esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan monooleate, and condensation products of these partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. The emulsion may also contain a sweetening agent and a flavoring agent. Syrups and elixirs can be formulated with a sweetening agent, such as glycerol, sorbitol, or sucrose. Such formulations may also contain a demulcent, a preservative, a flavoring agent, or a coloring agent.

[0105] The pharmaceutical composition of the present disclosure may be in the form of a sterile injectable preparation, for example, a sterile injectable aqueous or oily suspension. This suspension can be formulated according to known techniques using the above-mentioned suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation can also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, such as a solution in 1,3-butanediol, or can be prepared as a lyophilized powder. Among the acceptable vehicles and solvents that can be used are water, Ringer's solution, and isotonic sodium chloride solution. Further, a sterile fixed oil may be conventionally used as a solvent or suspension medium. For this purpose, any brand of fixed oil containing synthetic mono- or diglycerides can be used. Further, fatty acids such as oleic acid can also be used in the preparation of injectables. Among the acceptable vehicles and solvents that can be used are water, Ringer's solution, isotonic sodium chloride solution, and hypertonic sodium chloride solution.

[0106] The amount of the ROCK2 inhibitor that can be combined with the carrier material to produce a single dosage form can vary depending on the host to be treated and the particular mode of administration. For example, a sustained release formulation intended for oral administration to humans may contain from about 1 to 1000 mg of the active material, formulated with a convenient amount of the carrier material that can vary from about 5 to about 95% (weight: weight) of the total composition. The pharmaceutical composition can be prepared to provide an easily measurable amount for administration. For example, an aqueous solution intended for intravenous infusion may contain from about 3 to 500 μg of the ROCK2 inhibitor per milliliter of solution so that a suitable volume of infusion can be carried out at a rate of about 30 mL / hour.

[0107] Formulations suitable for topical administration to the eye also include eye drops in which the ROCK2 inhibitor is dissolved or suspended in a suitable carrier for the ROCK2 inhibitor, particularly an aqueous solvent. The ROCK2 inhibitor may be present in such formulations at a concentration of 0.5 to 20%, preferably 0.5 to 10%, particularly about 1.5% w / w.

[0108] Formulations suitable for topical administration to the mouth include lozenges having a flavor base, usually containing a ROCK2 inhibitor in sucrose and acacia or tragacanth; aromatic tablets containing a ROCK2 inhibitor in an inert base such as gelatin and glycerin, or sucrose and acacia; and mouthwashes containing a ROCK2 inhibitor in a suitable liquid carrier.

[0109] Formulations for rectal administration can be presented, for example, as suppositories containing a suitable base including cocoa butter or salicylate.

[0110] Formulations suitable for intrapulmonary or nasal administration have a particle size in the range of, for example, 0.1 to 500 microns, such as 0.5, 1, 30, 35, etc., and are administered by rapid inhalation through the nasal cavity or by inhalation through the mouth to reach the alveolar sac. Suitable formulations include aqueous or oily solutions of the ROCK2 inhibitor. Formulations suitable for aerosol or dry powder administration can be prepared according to conventional methods and can be delivered together with other therapeutic agents such as compounds.

[0111] Formulations suitable for vaginal administration can be presented as suppositories, pessaries, tampons, creams, gels, pastes, foams or spray formulations containing, in addition to the ROCK2 inhibitor, a carrier known in the art to be appropriate.

[0112] Formulations suitable for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain antioxidants, buffers, bacteriostatic agents and solutes rendering the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may contain suspending and thickening agents.

[0113] The formulation is presented in unit dose or multiple dose containers, such as sealed ampoules and vials, and can be stored in a freeze-dried (lyophilized) state that requires only the addition of a sterile liquid carrier, such as water for injection, immediately prior to use. Immediate injection solutions and suspensions are prepared from sterile powders, granules and tablets of the previously described kind. Preferred unit dosage formulations contain a daily dose or a sub-unit daily dose of a ROCK2 inhibitor, or an appropriate fraction thereof, as described hereinabove.

[0114] The present disclosure further provides a veterinary composition comprising at least one ROCK2 inhibitor as defined above, together with a veterinary carrier therefor.

[0115] The veterinary carrier is a material useful for the purpose of administering the composition and is otherwise inert or a solid, liquid or gaseous material acceptable in the veterinary art and compatible with the ROCK2 inhibitor. These veterinary compositions can be administered orally, parenterally, or by any other desired route.

[0116] Using the ROCK2 inhibitors of the present disclosure, controlled release pharmaceutical formulations ( "controlled release formulations") containing one or more of the ROCK2 inhibitors of the present disclosure as active ingredients can be provided, in which the release of the ROCK2 inhibitor is controlled and regulated so as to be able to reduce the dosing frequency or improve the pharmacokinetics or toxicity profile of a given ROCK2 inhibitor. Combination with other active agents

[0117] The ROCK2 inhibitors, such as the ROCK2 selective inhibitors, disclosed herein can be used in combination with at least one additional therapeutic agent. The at least one additional therapeutic agent can be, for example, an antiviral agent, a corticosteroid, an anti-inflammatory signaling modulator, a β2 adrenergic receptor agonist bronchodilator, an anticholinergic agent, a mucolytic agent, hypertonic saline, or a mixture thereof.

[0118] The ROCK2 inhibitor can be administered in combination with one or more additional antiviral therapies and / or antiviral agents. At least one other antiviral agent may be a nucleoside or nucleotide analog, or a pharmaceutically acceptable salt or prodrug thereof. The antiviral agent can be selected from remdesivir, ribavirin, favipiravir, T-705 monophosphate, T-705 diphosphate, T-705 triphosphate, ST-193, idoxuridine, edoxudine, trifluridine, vidarabine, brequinar, acyclovir, ganciclovir, valacyclovir, cidofovir, valganciclovir, penciclovir, famciclovir, zidovudine, didanosine, zalcitabine, stavudine, abacavir, lamivudine, emtricitabine, tenofovir disoproxil fumarate, tenofovir alafenamide fumarate, adefovir, entecavir, telbivudine, sofosbuvir, and combinations or mixtures thereof.

[0119] Also, for simultaneous or sequential administration to a patient, it is possible to combine any ROCK2 inhibitor of the present disclosure with one or more additional therapeutic agents in a single dosage form. The combination therapy can be administered as a simultaneous or sequential regimen. When administered sequentially, the combination can be administered in two or more administrations.

[0120] Co-administration of the ROCK2 inhibitor of the present disclosure with one or more other active therapeutic agents generally refers to simultaneous or sequential administration of the ROCK2 inhibitor of the present disclosure and one or more other active therapeutic agents such that a therapeutically effective amount of both the ROCK2 inhibitor of the present disclosure and one or more other active therapeutic agents are present in the patient's body.

[0121] Co-administration includes administration of a unit dose of a ROCK2 inhibitor of the present disclosure before or after administration of a unit dose of one or more additional therapeutic agents, e.g., administration of a unit dose of a ROCK2 inhibitor of the present disclosure within seconds, minutes, or hours of administration of one or more additional therapeutic agents. For example, after initially administering a unit dose of a ROCK2 inhibitor of the present disclosure, a unit dose of one or more additional therapeutic agents can be administered within seconds or minutes. Alternatively, after initially administering a unit dose of one or more additional agents, a unit dose of a ROCK2 inhibitor of the present disclosure can be administered within seconds or minutes. In some cases, it may be desirable to administer a unit dose of one or more additional therapeutic agents hours (e.g., 1 to 12 hours) after initially administering a unit dose of a ROCK2 inhibitor of the present disclosure. In other cases, it may be desirable to administer a unit dose of a ROCK2 inhibitor of the present disclosure hours (e.g., 1 to 12 hours) after initially administering a unit dose of one or more additional therapeutic agents.

[0122] Combination therapies can result in a “synergistic effect” and be “synergistic,” i.e., the effect achieved when the active ingredients are used together is higher than the sum of the effects obtained by using the compounds separately. A synergistic effect can be realized when the active ingredients are (1) co-formulated and administered or simultaneously delivered in a combined formulation; (2) delivered alternately or in parallel as separate formulations; or (3) delivered by some other regimen. When delivered by an alternating therapy, a synergistic effect can be realized when the active ingredients are administered or delivered sequentially, e.g., by different injections with separate tablets, pills or capsules, or separate syringes. Generally, during an alternating therapy, the effective dosage of each active ingredient is administered sequentially, i.e., continuously, whereas in a combination therapy, the effective dosages of two or more active ingredients are administered together. A synergistic antiviral effect means an antiviral effect that is higher than the predicted purely additive effect of the individual active ingredients of the combination. Method of treating viral infection

[0123] In one aspect, the present disclosure provides a method for treating viral infections, particularly coronavirus infections. In some embodiments, the method includes administering to a subject suffering from a viral infection one or more ROCK2 inhibitors, including the ROCK2 selective inhibitor of the present disclosure. The method may include administering a therapeutically effective amount of the ROCK2 selective inhibitor. In some embodiments, the method includes administering one or more ROCK2 inhibitors of the present disclosure to a subject at risk of viral infection. The method may include administering a prophylactically effective amount of the ROCK2 selective inhibitor.

[0124] In one aspect, the present disclosure provides a method for treating and preventing sequelae resulting from viral infections, including sequelae resulting from coronavirus infections. In some embodiments, the method includes administering to a subject suffering from a viral infection one or more ROCK2 inhibitors, including the ROCK2 selective inhibitor of the present disclosure. The method may include administering a therapeutically effective amount of the ROCK2 selective inhibitor.

[0125] In one aspect, the present disclosure provides a method for preventing viral infection in a subject at risk of viral infection, including the risk of coronavirus infection. In some embodiments, the method includes administering to a subject at risk of viral infection one or more ROCK2 inhibitors, including the ROCK2 selective inhibitor of the present disclosure. The method may include administering a prophylactically effective amount of the ROCK2 selective inhibitor.

[0126] As used herein, the term "administering" or "administration" of a ROCK2 inhibitor to a subject refers to contacting the ROCK2 inhibitor with the subject, or a cell, tissue, organ, or biological fluid of the subject. Such administration includes any route by which the ROCK2 inhibitor is introduced or delivered to perform its intended function. Administration can be effected by any route suitable for delivery of the ROCK2 inhibitor. Such administration can be carried out using one of a variety of methods known to those of ordinary skill in the art. For example, the ROCK2 inhibitors of the present disclosure can be administered systemically or locally. Thus, the delivery route may include oral, inhalation, transdermal, intravenous, intramuscular, intraperitoneal, or subcutaneous delivery. Administration includes self-administration and administration by another person. Administration can also be carried out, for example, once, multiple times, and / or over one or more extended periods. In some embodiments, administration includes both direct administration (including self-administration) and indirect administration, including the act of prescribing the drug.

[0127] As used herein, the terms "treating," "treatment," and "treat" refer to the administration of a therapeutic agent, such as a ROCK2 inhibitor disclosed herein or any composition containing such a ROCK2 inhibitor, internally or externally to a subject or patient having one or more disease symptoms, suspected of having a disease, or in which a therapeutic agent has therapeutic activity. "Treating," "treatment," and "treat" refer to therapeutic treatment and / or prophylactic treatment. These terms encompass the treatment of viral infections as described herein in a subject such as a human and include (i) inhibiting a viral infection, i.e., arresting its development; (ii) alleviating a viral infection, i.e., causing regression of the infection; (iii) slowing the progression of a viral infection; and / or (iv) inhibiting, alleviating, or slowing the progression of one or more symptoms of a viral infection. Treating and treatment refer to the intentional act of a physiological intervention intended to cure, delay, or ameliorate one or more symptoms associated with a viral infection. When treatment is administered prior to the clinical manifestations of a viral infection or detection of a viral infection, the treatment is considered prophylactic. Reduction or alleviation of symptoms of a viral infection can be evaluated by any clinical measurement typically used by a physician or other skilled artisan to assess the severity or progression of such symptoms. The terms further refer to the delaying of the onset of one or more symptoms and / or a decrease in the severity of one or more symptoms associated with a viral infection. The term further includes ameliorating existing uncontrolled or undesirable symptoms, preventing additional symptoms, and ameliorating or preventing the root cause of such symptoms. Thus, the term means that a beneficial result has been brought about in the subject.

[0128] As used herein, the terms "prevent" and "prevention" refer to acting prophylactically prior to a viral infection, preventing the onset of a viral infection, or minimizing the extent of a viral infection or delaying the course of its onset.

[0129] As used herein, the terms "patient," "subject," "individual," etc. are used interchangeably herein and refer to any animal suitable for the methods described herein. In some embodiments, the patient, subject or individual is a human. In some embodiments, the subject is a veterinary subject. In some embodiments, the subject is a mammal. In some embodiments, the subject is a companion animal. In some embodiments, the subject is a livestock animal. In some embodiments, the subject is a laboratory animal. In some embodiments, the subject is a cat, dog, gorilla, mink, ferret, lion, tiger, puma, cougar, snow leopard, bat, hamster, pig, rabbit, raccoon, muskrat, deer, rhesus monkey, cynomolgus monkey, baboon, green monkey, common marmoset, mouse, and rat. In some embodiments, the subject is a mink.

[0130] As used herein, the terms "effective amount" or "therapeutically effective amount" with respect to the ROCK2 inhibitors of the present disclosure refer to an amount sufficient to provide a therapeutic benefit in a patient after administration, e.g., to improve one or more symptoms of a viral infection in a subject, or to delay, reduce, minimize, alleviate, or ameliorate the symptoms of a viral infection compared to an untreated patient. The effective amount may vary depending on the species, age, weight, health of the subject and the nature or severity of the viral infection. Depending on the mode of administration, the effective amount may likewise vary. In some cases, multiple doses of the ROCK2 inhibitor are administered to achieve an effective amount for the intended therapeutic benefit. The therapeutically effective amount is also such that any toxic or adverse effects of the antibody are outweighed by the therapeutically beneficial effects. Determining the therapeutically effective amount of the antibodies disclosed herein based on these factors is routine in the art for one of ordinary skill in the art. "Preventive effective amount" refers to the dosage required to achieve the desired preventive result and an amount effective over a period of time. Typically, a prophylactic dose is used prior to viral infection or in a subject at an early stage of viral infection, and thus the preventive effective amount may be less than the therapeutically effective amount.

[0131] In embodiments, the viral infection is caused by a virus selected from the group consisting of SARS-CoV-1, SARS-CoV-2, MERS-CoV, yellow fever, eastern equine encephalitis virus, human immunodeficiency virus (HIV), African swine fever virus, alphavirus, adenoviridae, arenaviridae, arterivirus, astroviridae, baculoviridae, birnaviridae, bunyaviridae, caliciviridae, calimovirus, sarcovirus, coronaviridae, cystovirus, EBV, deltavirus, filoviridae, phenuiviridae, flaviviridae, iridovirus, mononegavirus, myoviridae, papillomavirus, papovaviridae, paramyxoviridae, prion, parvoviridae, phycodnaviridae, poxviridae, potyviridae, reoviridae, retroviridae, rhabdoviridae, tectivirus, togaviridae, pox, papilloma, influenza, Sendai virus (SeV), Sindbis virus (SINV), vaccinia virus, West Nile, hantavirus, cold-causing viruses, and any combination thereof. In embodiments, the viral infection is caused by a virus selected from the group consisting of yellow fever, eastern equine encephalitis virus, human immunodeficiency virus (HIV), African swine fever virus, alphavirus, adenoviridae, arenaviridae, arterivirus, astroviridae, baculoviridae, birnaviridae, bunyaviridae, caliciviridae, calimovirus, sarcovirus, coronaviridae, cystovirus, EBV, deltavirus, filoviridae, phenuiviridae, flaviviridae, iridovirus, mononegavirus, myoviridae, papillomavirus, papovaviridae, paramyxoviridae, prion, parvoviridae, phycodnaviridae, poxviridae, potyviridae, reoviridae, retroviridae, rhabdoviridae, tectivirus, togaviridae, pox, papilloma, influenza, Sendai virus (SeV), Sindbis virus (SINV), vaccinia virus, West Nile, hantavirus, cold-causing viruses, and any combination thereof.In embodiments, the viral infection is caused by coronaviruses such as SARS-CoV-1, SARS-CoV-2, and MERS-CoV. In embodiments, the viral infection is caused by SARS-CoV-1. In embodiments, the viral infection is caused by SARS-CoV-2. In some embodiments, the viral infection is caused by the delta or omicron variant of SARS-CoV-2. In embodiments, the viral infection is caused by MERS-CoV.

[0132] In embodiments, the present disclosure provides a method for treating or preventing one or more sequelae of COVID-19, the method comprising administering to a subject in need thereof a therapeutically effective amount of a ROCK2 inhibitor, which comprises a ROCK2 selective inhibitor of the present disclosure. In some embodiments, the ROCK2 inhibitor is an RNA, such as an antisense RNA, siRNA, or miRNA. In some embodiments, the ROCK2 inhibitor is a compound of Formula I-IV as defined herein, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotope-labeled compound thereof. In embodiments, the present disclosure provides a method for treating or preventing one or more sequelae of COVID-19, the method comprising administering to a subject in need thereof a therapeutically effective amount of Compound 1 as defined herein, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotope-labeled compound thereof. In some embodiments, the present disclosure provides a method for treating or preventing one or more sequelae of COVID-19, the method comprising administering to a subject in need thereof a therapeutically effective amount of belumosudil.

[0133] The sequelae of coronavirus infection, particularly SARS-CoV-1, SARS-CoV-2 (including all variants such as Delta and Omicron), and MERS-CoV, can result from one or more phenomena including organ damage derived from the acute infection phase, signs of a persistent hyperinflammatory state, ongoing viral activity associated with the host-virus reservoir, or an insufficient antibody response. Sequelae of COVID-19 include, but are not limited to, fatigue, dyspnea (shortness of breath), cough, arthralgia (joint pain), and chest pain. Further sequelae include cognitive impairment, depression, myalgia, headache, fever, and palpitations. The sequelae of COVID-19 may be cardiovascular (e.g., myocarditis, ventricular dysfunction, stroke), respiratory (e.g., lung function abnormalities, fibrosis), renal (e.g., acute kidney injury), skin (e.g., rash, alopecia), neurological (e.g., olfactory and gustatory dysfunction, abnormal sleep regulation, cognitive changes, memory impairment), and / or psychiatric (e.g., depression, anxiety, mood swings). As a result of treating one or more sequelae of COVID-19, ROCK2 selective inhibitors may be useful for treating or preventing long COVID.

[0134] Without being bound by theory, ROCK2 inhibitors such as, for example, ROCK2 selective inhibitors, including antisense RNA, siRNA or miRNA molecules and compounds of formulas I-IV and compound 1, can block (1) the pathways used by the virus to enter cells; (2) the cytoskeleton used by the virus for migration and spread; (3) the pathways used by the virus to upregulate the energy metabolism of cells; and (4) the pathways used by the virus to spread to other cells.

[0135] The ROCK2 inhibitors provided herein, including the ROCK2 selective inhibitor, can prevent the interaction of the virus with the cytoskeleton of the host cell, thereby inhibiting virus entry, replication, and / or spread. The cytoskeleton is a complex network in eukaryotic cells, which includes three major types of cytoskeletal polymers (including actin filaments, microtubules, and intermediate filaments), and enables the cell to perform multiple functions in an integrated manner, such as connecting to the external environment, moving, adjusting the forces for shape change, transporting vesicles through the cytoplasm, and spatially organizing the contents. Most viruses hijack one or more aspects of the cytoskeletal network to facilitate their own infection. The interaction of the virus with the actin filaments, microtubules, and intermediate filaments of the host cell plays an important role in the life cycle of the virus, especially the coronavirus.

[0136] The ROCK2 inhibitors provided herein, including the ROCK2 selective inhibitor, can prevent the entry of the virus into its target cells. After binding to the target cells, the virus can migrate to a preferred site for entry. When the virus reaches the entry site, it has been observed that the actin filaments contract and concentrate around the plasma membrane. Pharmacological stabilization of the actin cortex can prevent virus penetration of the host cell. Viruses also utilize cytoskeleton-regulated signaling pathways as part of their infection process.

[0137] Coronaviruses can utilize three cytoskeletal networks to complete the virus transport process. Transport from / to the cell periphery for short-distance pathways is mediated by its motor proteins such as actin and myosin, while long-distance transport is mediated by microtubules and the motor proteins dynein and kinesin. After entering the host cell, the vesicles containing coronaviruses move along the microtubules and move from the plasma membrane towards the replication site.

[0138] SARS-CoV-2-infected cells can fuse with neighboring cells to form actin-regulated syncytia. During the infection stage, SARS-CoV-2 surfs along the filopodia on the host membrane towards the cell entry site, uses intermediate filament proteins to assist and utilize the angiotensin-converting enzyme-2 (ACE2) receptor, and enters the target cells. In a mouse SARS model, the SARS-CoV spike protein bound to the ACE2 receptor, reduced ACE2 expression, and caused severe lung injury. By binding to the ACE2 receptor to gain cell entry, SARS-CoV-2 efficiently blocks ACE2 activity and the conversion of AT-II to angiotensin, leading to vasoconstriction, increased vascular permeability, harmful myocardial remodeling, and acute lung injury. Downregulation of ACE2 and disruption of the normal feedback loop can also result in a life-threatening cytokine storm, as commonly observed in moderate to severe cases of COVID-19. The excessive inflammatory response to SARS-CoV-2 is a major cause of disease severity and death in COVID-19 patients.

[0139] When ROCK2 is inhibited, ACE2 activity is restored, AT-II conversion continues, and inflammatory cytokines (i.e., IL-1β, IL-6, and TNF-alpha), TGF-β1, and fibrosis-forming markers (i.e., COL1A1, COL3A1, alpha-SMA, fibronectin, CTGF, and PDGF-B) are downregulated. ROCK inhibition may also play a role in preventing viral spread between host cells by inhibiting Rho GTPase-mediated cytoskeletal reorganization, including cell migration, invasion, survival, cell death, and the formation of viral syncytia.

[0140] Furthermore, the neurogenic disorders and respiratory tract injuries caused by the coronavirus are at least partially microtubule-dependent. Structural damage to the respiratory epithelium and ciliary dysfunction are typical pathological conditions of coronavirus infection. Cilia are complex structures present on the cell surface composed of microtubules. Coronaviruses that cause severe airway damage can do so by causing ciliary loss in the upper airway and lungs. Furthermore, disruption of microtubules can be associated with neurodegenerative diseases.

[0141] Based on its multifaceted mode of action, the ROCK2 inhibitors, including the ROCK2-selective inhibitors of the present disclosure, such as antisense RNAs, siRNAs, or miRNAs of ROCK2 transcripts and the compounds of Formulas I-IV and Compound 1 as defined herein, can inhibit or prevent viral cell entry and inhibit viral spread, for example, by inhibiting syncytia. Second, ROCK2 inhibition also alleviates the overactivation of the immune response known as the "cytokine storm" and the fibrotic changes in blood vessels, heart, and lung tissues that have resulted in long-term sequelae in some COVID-19 patients.

[0142] In some embodiments, the ROCK2 inhibitor inhibits and / or modulates viral entry of a viral infection.

[0143] In some embodiments, the ROCK2 inhibitor inhibits and / or modulates viral spread of a viral infection.

[0144] In some embodiments, the ROCK2 inhibitor targets a cellular target of a subject. Since the ROCK2 inhibitor interacts with a host cellular target rather than a viral target, the development of drug resistance to the effect of the ROCK2 inhibitor in the virus can be slowed or substantially absent compared to other antiviral agents or compositions. In some embodiments, the use of the ROCK2 inhibitor for the treatment of viral diseases is characterized by the absence of drug resistance.

[0145] In some embodiments, the ROCK2 inhibitor inhibits and / or modulates the formation of stress fibers.

[0146] In some embodiments, the ROCK2 inhibitor inhibits and / or modulates the dynamics of actin filaments.

[0147] In some embodiments, the ROCK2 inhibitor inhibits and / or modulates the mammalian target of rapamycin (mTOR) metabolic pathway.

[0148] In some embodiments, the ROCK2 inhibitor inhibits and / or modulates the AKT (serine / threonine protein kinase B) metabolic pathway.

[0149] In some embodiments, the ROCK2 inhibitor inhibits and / or modulates the fusion pathway.

[0150] In some embodiments, the subject is a human subject. In some embodiments, the subject is a veterinary subject. In some embodiments, the human subject is an adult patient. In some embodiments, the human subject has or is at risk of having fibrosis (including post-COVID fibrosis) or scarring in the lung. In some embodiments, the human subject is a pediatric patient. In some embodiments, the human subject has or is at risk of developing Kawasaki disease, fibrosis, or scarring in the lung.

[0151] In embodiments, the ROCK2 inhibitor at least partially reverses and / or inhibits the level of fibrosis, at least partially inhibits the excessive deposition of extracellular matrix in the lung, or improves blood supply to the lung. In some embodiments, the ROCK2 selective inhibitor at least partially reverses and / or inhibits the level of post-COVID fibrosis. In some embodiments, administration of the ROCK2 inhibitor results in at least one of the following: (a) at least a 5% reduction in pulmonary edema; (b) at least a 5% reduction in the pulmonary pathology severity score associated with pulmonary fibrosis; (c) at least a 5% reduction in the expression of pro-inflammatory proteins; or (d) at least a 5% reduction in the expression of fibrotic proteins.

[0152] In some embodiments, the subject experiences a decrease in forced vital capacity of (a) less than 10%; (b) less than 9%; (c) less than 8%; (d) less than 7%; (e) less than 6%; (f) less than 5%; (g) less than 4%; (h) less than 3%; (i) less than 2%; or (j) less than 1% after administration of the ROCK2 inhibitor to the subject for at least two weeks. In some embodiments, the subject does not experience a decrease in forced vital capacity after administration of the ROCK2 inhibitor to the subject for at least two weeks. In some embodiments, the subject experiences an increase in forced vital capacity of (a) at least 0.5%; (b) at least 1%; (c) at least 1.5%; (d) at least 2%; (e) at least 2.5%; (f) at least 3%; (g) at least 3.5%; (h) at least 4%; (i) at least 4.5%; or (j) at least 5% after administration of the ROCK2 inhibitor to the subject for at least two weeks.

[0153] In some embodiments, the subject experiences a reduction in the development of coronary artery lesions at one month of illness of (a) at least 5%; (b) at least 10%; (c) at least 15%; (d) at least 20%; (e) at least 25%; (f) at least 30%; (g) at least 35%; or (h) at least 40% after administration of the ROCK2 inhibitor to the subject for at least two weeks. In some embodiments, the ROCK2 inhibitor at least partially reverses or reduces a vascular inflammatory syndrome, or at least partially reverses or reduces a rash, redness of the eyes, lips or tongue, swelling of the hands or feet, redness of the hands or feet, or swelling of the neck. In some embodiments, the ROCK2 inhibitor restores the balance of the immune response in the subject.

[0154] In one aspect, the present disclosure provides a method of treating a patient suffering from a viral disease, the method comprising administering to a patient in need of such treatment a therapeutically effective amount of a ROCK2 inhibitor, such as a ROCK2 selective inhibitor of the present disclosure. In some embodiments, the ROCK2 inhibitor is an RNA. In some embodiments, the ROCK2 inhibitor is an antisense RNA against ROCK2 transcription. Optionally, the RNA is a small interfering RNA (siRNA) or a microRNA (miRNA). In some embodiments, the ROCK2 inhibitor is a compound having a structure of Formulas I-IV as defined herein, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotope-labeled compound thereof, particularly (2-(3-(4-((1H-indazol-5-yl)amino)quinazolin-2-yl)phenoxy)-N-isopropylacetamide). In some embodiments, the ROCK2 inhibitor has a structure of Compound 1 as defined herein, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotope-labeled compound thereof. In some embodiments, the ROCK2 inhibitor is belumosudil.

[0155] One or more ROCK2 inhibitors, including the ROCK2 selective inhibitors of the present disclosure, are administered by any route suitable for the condition to be treated. Suitable routes include oral, rectal, nasal, pulmonary, topical (including buccal and sublingual), vaginal, and parenteral (including subcutaneous, intramuscular, intravenous, intradermal, intrathecal, and epidural). It can be understood that the preferred route may vary, for example, with the condition of the recipient. An advantage of the compounds of the present disclosure is that they are orally bioavailable and can be dosed orally.

[0156] The effective dosage of the compounds of the present disclosure depends at least on the nature of the condition being treated, the toxicity, whether the compound is being used prophylactically or against active viral infection, the method of delivery, and the pharmaceutical formulation, and can be determined by a clinician using conventional dose escalation studies. In embodiments, the dosage of the compounds of the present disclosure ranges from about 0.1 to about 50 mg / kg body weight per day. The daily dosage for an adult can range between 1 mg and 1000 mg, for example, between about 5 mg and about 800 mg or between about 50 mg and 500 mg, and can be in the form of a single or multiple doses per day. In embodiments, the daily dosage of the ROCK2 inhibitor for treating a viral infection or treating or preventing one or more sequelae resulting from the infection is about 10 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, about 600 mg, about 650 mg, about 700 mg, about 750 mg, or about 800 mg, which can be administered as a single daily dose or divided between two, three, or more administrations per day. In embodiments, the dosage is about 400 mg per day BID. Method of treating symptoms associated with Kawasaki disease / viral infection

[0157] Among pediatric patients infected with COVID-19, some exhibit Pediatric Multisystem Inflammatory Syndrome (PMIS), or Pediatric Inflammatory Multisystem Syndrome Temporally Associated with SARS-CoV-2 (PIMS-TS), which resembles an inflammatory disease called Kawasaki disease. As used herein, the term "Kawasaki disease" generally refers to a vasculitis syndrome or inflammation of blood vessels, sometimes an inflammatory disease causing swelling throughout the body, including Kawasaki disease-like diseases such as PMIS or PIMS-TS. Symptoms of Kawasaki disease may include persistent high fever (above 101°F) for at least 4 days, in addition to rash, redness of the eyes, lips / tongue, swelling and redness of the hands / feet, and swelling of the neck. As used herein, the term "Kawasaki disease shock syndrome" or KDSS refers to a Kawasaki disease patient showing a decrease of more than 20% in systolic blood pressure compared to healthy subjects of the same age, or a patient showing peripheral blood circulation disorder. KDSS may be caused by high levels of circulating inflammatory factors.

[0158] Among pediatric patients with Kawasaki disease associated with COVID-19, PMIS, or PIMS-TS, there may be an increase in inflammatory markers, including, for example, neutrophilia, C-reactive protein (CRP), procalcitonin (PCT), and IL-6. The onset of Kawasaki disease or PMIS associated with COVID-19 may be caused by a post-viral infection immune response, such as an antibody or immune complex-mediated reaction.

[0159] As a form of systemic vasculitis, Kawasaki disease can involve small to medium-sized blood vessels. The most severe complications or sequelae may be the formation of coronary artery lesions (CAL) such as myocardial infarction, coronary artery fistula, coronary artery dilation, and coronary artery aneurysm, which can subsequently lead to long-term sequelae such as stenosis or occlusion and myocardial infarction. Therefore, a decrease in the ratio of coronary artery aneurysms among Kawasaki disease patients can be an indicator of the effectiveness of treatment.

[0160] The ROCK2 inhibitors of the present disclosure, including antisense RNAs, siRNAs or miRNA molecules, or compounds of Formulas I-IV defined herein, or Compound 1, etc., which are ROCK2 selective inhibitors, can reduce inflammatory cytokines, such as the peripheral blood levels of IL-17 and IL-23. Further, these ROCK2 inhibitors can restore the balance of the immune response in pediatric patients suffering from Kawasaki disease, PMIS, or PIMS-TS after infection with COVID-19 or other coronaviruses. A method of treating fibrosis caused by viral infection

[0161] COVID-19 patients can develop scarring in their lungs during the acute phase of the disease or after the patient has recovered from the illness. The scarring in the lungs may be fibrosis (including post-COVID fibrosis) or fibrotic scarring. The underlying cause and clinical course of lung scarring in COVID-19 patients may be the same as or different from those due to interstitial lung diseases and pulmonary fibrosis, such as idiopathic pulmonary fibrosis (IPF) or rheumatoid arthritis.

[0162] Fibrosis is the overgrowth, hardening, and / or scarring of various tissues and can be caused by the excessive deposition of extracellular matrix components, such as collagen. Fibrosis can be the result of a chronic inflammatory response induced by various stimuli, including persistent infection, autoimmune reactions, allergic responses, chemical injury, radiation, and tissue injury.

[0163] COVID-19 infection can result in a variety of respiratory diseases ranging from atypical pneumonia to acute respiratory distress syndrome (ARDS). Many patients infected with the virus may share the characteristics of patients suffering from IPF. For IPF patients, lung function deteriorates relentlessly, leading to respiratory failure and death. A possible cause of lung injury can be cytokine release syndrome induced by viral antigens.

[0164] The ROCK2 signaling pathway controls cell movement and shape. Furthermore, ROCK2 can regulate cytokine secretion in T cells, such as by promoting inflammatory cytokines such as IL-17 and IL-21. However, the secretion of anti-inflammatory cytokines IL-2 and IL-10 is negatively regulated by ROCK2 under Th17 skewing activation. Also, in diseases rather than under steady-state conditions, ROCK2 contributes to the regulation of IFN-γ secretion in T cells derived from rheumatoid arthritis patients. Thus, ROCK2 signaling is an important pathway in the modulation of T cell-mediated immune responses that emphasizes the therapeutic potential of targeted inhibition of ROCK2 in autoimmunity. Therefore, ROCK2 inhibitors can downregulate the secretion of IL-21 and IL-17 in human T cells by a STAT3-dependent mechanism.

[0165] The ROCK2 inhibitors of the present disclosure, including ROCK2-selective inhibitors such as antisense RNA, siRNA or miRNA molecules or compounds of formulas I-IV or Compound 1 as defined herein, can reduce the peripheral blood levels of two inflammatory cytokines, IL-17 and IL-23. Furthermore, these ROCK2 inhibitors can simultaneously upregulate the immunosuppressive cytokine IL-10 and increase the percentage of Foxp3+CD4 T cells in the blood, which can reduce the immune-inflammatory response. Thus, the ROCK2 inhibitors of the present disclosure, including ROCK2-selective inhibitors such as antisense RNA, siRNA or miRNA molecules or compounds of formulas I-IV or Compound 1 as defined herein, can restore the balance of the immune response in patients suffering from fibrosis after infection with SARS-CoV-2 or other coronaviruses. Furthermore, the ROCK2 inhibitors of the present disclosure, including ROCK2-selective inhibitors such as antisense RNA, siRNA or miRNA molecules or compounds of formulas I-IV or Compound 1 as defined herein, can treat or prevent pulmonary fibrosis, including post-COVID fibrosis.

[0166] Using several pulmonary function parameters, an effective amount of the ROCK2 inhibitor of the present disclosure can be determined, i.e., an amount that reduces, stabilizes, or reverses the pathological reduction rate of one or more pulmonary function parameters in a patient suffering from a viral infection such as a coronavirus infection; or the response of a patient to ROCK2 inhibitor therapy can be monitored. These pulmonary function parameters may include vital capacity (VC), forced expiratory volume (FEV), forced vital capacity (FVC), and FVC%.

[0167] As used herein, the term "vital capacity" (VC) refers to the total volume of air that can enter and exit the lungs. VC is equal to the sum of the inspiratory reserve volume, the tidal volume, and the expiratory reserve volume.

[0168] As used herein, the term "forced expiratory volume" (FEV) refers to a measurement of how much air a subject can exhale during forced breathing. The amount of air exhaled can be measured at the 1st second (FEV1), 2nd second (FEV2), and / or 3rd second (FEV3) of forced breathing. For example, the FEV1 / FVC ratio refers to the ratio of the forced expiratory volume in one second to the forced vital capacity.

[0169] As used herein, the term "forced vital capacity" (FVC) refers to the vital capacity resulting from a maximal forced expiratory effort, i.e., the total amount of air exhaled and released by a subject during an FEV test.

[0170] As used herein, the term "FVC%" refers to the percent change in FVC of a subject over a period of time. The predicted FVC% is the measured FVC of the subject expressed as a percentage of the predicted FVC for the subject. As used herein, all FVC% predicted values are absolute values, not relative values.

[0171] Many of these pulmonary function parameters can be easily obtained using a spirometer. Residual volume can be obtained by indirect methods such as radiometric area measurement, body plethysmography, closed-circuit dilution methods (including helium dilution techniques), and nitrogen washout methods.

[0172] Vital capacity and related pulmonary function parameters naturally decline with aging. A number of studies have been conducted among normal populations to determine the rate of decline of vital capacity and other pulmonary function parameters. See Crapo et al. (1981) Am. Rev. Respir. Dis. 123:659-664. For example, a 65-year-old white male is expected to have a 0.03 liter decrease in FVC at age 66.

[0173] In contrast to the natural decline due to aging, subjects having a lung disease such as pulmonary fibrosis and fibrosis caused by COVID-19 have an abnormally rapid rate of decline, i.e., a "pathological rate of decline", in vital capacity or one or more lung function parameters. As used herein, a "pathological rate of decline" is a rate of decline in vital capacity or one or more lung function parameters that is at least 5% higher than the decline due to normal aging. In some embodiments, the pathological rate of decline is at least 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 200%, 300%, 400%, 500%, 600%, 700%, 800% or 1000% higher than the predicted rate of decline of a normal person of a similar and matched race or ethnicity, gender, age, height, and weight. The rate of decline can be expressed as a change from baseline per week, two weeks, four weeks, eight weeks, twelve weeks, twenty-four weeks, thirty-six weeks, forty-eight weeks, or twelve months. In some embodiments, the pathological rate of decline in vital capacity is a change in forced vital capacity (FVC) from baseline of at least about -0.05 liters, -0.10 liters, -0.15 liters, -0.20 liters or -0.25 liters per twelve months. In other embodiments, the pathological rate of decline is a change from the predicted baseline forced vital capacity percent (FVC%) of at least about -2%, -3%, -4%, -5%, -6%, -7%, -8% or -10% per twelve months.

[0174] Administration of an ROCK2 inhibitor of the present disclosure, including an antisense RNA, siRNA or miRNA molecule, or a compound of Formula I-IV as defined herein, or a compound such as Compound 1, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotopically labeled compound thereof, can result in an increase in FVC in a subject having fibrosis after treatment compared to before treatment. Treatment with an effective amount of an ROCK2 inhibitor can increase FVC by at least 0.5%, 1%, 1.5%, 2.0%, 2.5%, 3.0%, 4.0%, 5.0%, 6.0%, 7.0%, 8.0%, 9.0%, 10%, 15%, 20%, 30%, 40% or 50% compared to pre-treatment FVC. In some embodiments, treatment with an ROCK2 inhibitor is for at least 1 week, 2 weeks, 3 weeks, 6 weeks, 9 weeks, 12 weeks, 15 weeks, 18 weeks, 21 weeks, 24 weeks, 27 weeks, 30 weeks, 33 weeks, 36 weeks or 48 weeks. In some embodiments, the treatment is 2 weeks or less, 3 weeks or less, 6 weeks or less, 9 weeks or less, 12 weeks or less, 18 weeks or less, 24 weeks or less, 36 weeks or less, 48 weeks or less, 12 months or less, 16 months or less, 20 months or less, or 24 months or less from the start of treatment with the ROCK2 inhibitor. Kit

[0175] In one aspect, the present disclosure provides a kit or composition comprising a ROCK2 inhibitor, including an antisense RNA, siRNA or miRNA molecule, or a compound of Formula I-IV or Compound 1 as defined herein, or a pharmaceutically acceptable salt, pharmaceutically acceptable ester, stereoisomer, mixture of stereoisomers or tautomer thereof. In some embodiments, individual kits are provided that include a compound of Formula I-IV or Compound 1 or a pharmaceutically acceptable salt, pharmaceutically acceptable ester, stereoisomer, mixture of stereoisomers or tautomer thereof as defined herein. In one aspect, the kit includes a compound of Formula I-IV or Compound 1 or a pharmaceutically acceptable salt thereof. Each of the individual kits described herein may optionally include a label and / or instructions for use of the compound in the treatment of a disease or condition in a subject (e.g., a human) in need thereof. In some embodiments, the disease or condition is a coronavirus infection. In other embodiments, each separate kit may also optionally include instructions for use of an additional pharmaceutical agent in combination with a ROCK2 inhibitor of the present disclosure, including an antisense RNA, siRNA or miRNA molecule, or a compound of Formula I-IV or Compound 1 as defined herein, in the treatment of a disease or condition in a subject (e.g., a human) in need thereof. In some embodiments, the kit includes belumosudil. In some embodiments, the kit includes individual dosage units of a ROCK2 inhibitor, including a ROCK2 selective inhibitor as described herein. Examples of individual dosage units may include pills, tablets, capsules, prefilled syringes or syringe cartridges, IV bags, etc., each containing a therapeutically effective amount of the ROCK2 inhibitor in question, or a pharmaceutically acceptable salt, racemate, enantiomer, diastereomer, tautomer, polymorph, pseudopolymorph, amorphous form, hydrate or solvate thereof. In some embodiments, the kit may contain a single dosage unit, or alternatively, where multiple dosage units are present, may contain, for example, the number of dosage units required for a particular regimen or period.

[0176] Also provided are the ROCK2 inhibitors of the present disclosure, including ROCK2 selective inhibitors such as antisense RNA, siRNA or miRNA molecules or compounds of Formulas I-IV or Compound 1 as defined herein or pharmaceutically acceptable salts, pharmaceutically acceptable esters, stereoisomers, mixtures of stereoisomers or tautomers thereof; and products comprising a container. In one aspect, the product comprises a compound of Formulas I-IV or Compound 1 or a pharmaceutically acceptable salt thereof, and a container. In some embodiments, the product comprises velmosdil. In some embodiments, the container of the product is a vial, jar, ampoule, pre-filled syringe, blister package, tin can, can, bottle, box, or intravenous bag. Exemplary embodiments

[0177] Certain embodiments of the invention are described in the following numbered paragraphs. 1. A method for treating viral infection in a subject in need of treatment for viral infection, the method comprising administering to the subject a therapeutically effective amount of a ROCK2 inhibitor. 2. A method for preventing viral infection in a subject at risk of viral infection, the method comprising administering to the subject a prophylactically effective amount of a ROCK2 inhibitor. 3. A method for treating Kawasaki disease, PMIS, or PIMS-TS associated with viral infection in a subject in need of treatment for Kawasaki disease, PMIS, or PIMS-TS associated with viral infection, the method comprising administering to the patient a therapeutically effective amount of a ROCK2 inhibitor. 4. A method for treating or preventing sequelae resulting from viral infection in a subject in need of treatment or prevention of sequelae resulting from viral infection, the method comprising administering to the patient a therapeutically effective amount of a ROCK2 inhibitor. 5. The method according to any one of paragraphs 1-4, wherein the ROCK2 inhibitor is a ROCK2 selective inhibitor. 6. The method according to any one of paragraphs 1-5, wherein the ROCK2 inhibitor is a ribonucleic acid (RNA). 7. The method according to paragraph 6, wherein the ROCK2 inhibitor is antisense RNA, small interfering RNA or microRNA. 8. The ROCK2 inhibitor has the structure of Formula I:

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Examples

[0178] The following examples are representative of embodiments of the compositions and methods described herein and are not meant to be limiting in any way. Example 1 in vitro SARS-CoV-2 prevention model

[0179] A 50 mM stock solution of Compound 2 (in DMSO) was diluted to concentrations of 100, 75, 50, 25, 10, 5, 1, 0.3, 0.1, and 0.03 μM using PBS. Approximately 30,000 immortalized human airway epithelial cells (Calu-3) (see Figures 1-10) or primary human airway epithelial cells (Normal Human Small Airway Epithelial Cells; Sigma) (see Figures 11-12) were grown to approximately 100,000 cells per well in a 96-well format. The cells were seeded in triplicate at 10 2 or 10 3 PFU of SARS-CoV-2 (standard CDC SARS-CoV-2 strain), incubated at 37 °C for 6 hours, and then washed. The cells were then treated with Compound 2 or the negative control by adding, in triplicate, 10 μL of a solution of Compound 2 (0.03 - 100 μM) or the negative control solution (DMSO in PBS) and incubating at 37 °C for 2 hours. The positive control consisted of remdesivir (RDV; 10 μM). Additional controls (3 wells) were treated with the highest concentration of Compound 2 but not infected with the virus. The cells were maintained until the second endpoint readout at 48 hours (the first was performed 24 hours post-infection). In some examples, the cell culture medium was replaced at 24 hours post-infection with either untreated culture medium (“unconditioned medium”) or culture medium containing the positive control (RDV) or Compound 2 (“conditioned medium”). See Figure 10.

[0180] Viral infection / viral load was analyzed by qPCR (qRT-PCR). Cellular RNA was extracted using TRIzol reagent (Thermo Fisher) according to the manufacturer's instructions. Viral or host RNA levels in the supernatant were determined using the following primers: Forward: 5’-GACCCCAAAATCAGCGAAAT-3’; Reverse: 5’-TCTGGTTACTGCCAGTTGAATCTG-3’ on a CFX Connect Real-Time System (Bio-Rad) instrument using TaqPath TM 1-Step RT-qPCR Master Mix (Thermo Fisher).

[0181] The toxicity of compound 2 was measured at concentrations of 0.1, 0.3, 1, 10, 25, 50, 75, and 100 μM in Calu-3 cells by LDH (lactate dehydrogenase) assay (Clontech) according to the manufacturer's instructions. The positive control consisted of saponin (0.5%). The negative control consisted of DMSO (0.2%).

[0182] The dataset was analyzed and plotted in GraphPad Prism (version 9.0.2). Analysis simulations of dose-response curves using the 50% effective principle and the law of mass action were performed using CompuSyn (https: / / www.combosyn.com / ). The results of the in vitro tests are provided in Figures 1 - 12, showing the ability of compound 2 to substantially reduce the viral titer in immortalized human airway epithelial cells infected with SARS-CoV-2, as well as its low cytotoxicity.

[0183] For example, FIGS. 1-3 show the in vitro efficacy of compound 2 (10 μM) in inhibiting virus replication 24 and 48 hours (FIG. 1), 24 hours (FIG. 2), and 48 hours (FIG. 3) after comparison with the control, indicating that the efficacy correlates with the administration time. FIGS. 4 and 5 show the low cytotoxicity of compound 2 at various concentrations (0.03, 0.1, 0.3, 1, 10, 25, 50, 75, and 100 μM) measured by LDH. FIGS. 6 and 7 show the in vitro efficacy of compound 2 (10 μM) in inhibiting virus replication 24 hours (FIG. 6) and 48 hours (FIG. 7) after comparison with the negative control (0.1% DMSO) and positive control (RDV). FIGS. 8A-8F show that the ability of various doses (1 μM, 5 μM, 10 μM, and 25 μM) of compound 2 to inhibit virus replication 24 hours after infection correlates with the administration time after infection - 0 hours (FIG. 8A); 1 hour (FIG. 8B); 2 hours (FIG. 8C); 4 hours (FIG. 8D); 8 hours (FIG. 8E) and 10 hours (FIG. 8F). FIGS. 9A-9F show that the ability of various doses (1 μM, 5 μM, 10 μM, and 25 μM) of compound 2 to inhibit virus replication 48 hours after infection correlates with the administration time after infection - 0 hours (FIG. 9A); 1 hour (FIG. 9B); 2 hours (FIG. 9C); 4 hours (FIG. 9D); 8 hours (FIG. 9E) and 10 hours (FIG. 9F). Dose responses were observed at all time points. FIG. 10 shows that repeated dosing of compound 2 further reduced the virus concentration. FIGS. 11 and 12 show that the efficacy of compound 2 at 5 μM and 25 μM concentrations in the treatment of SARS-CoV-2 infection in primary human airway cells 24 hours (FIG. 11) and 48 hours (FIG. 12) after infection correlates with the administration time after infection. Example 2 Compound 2 reduces the spread of SARS-CoV-2 in the lungs in vivo.

[0184] Human ACE2 transgenic mice (n = 5 or n = 8 per group) from Jackson Laboratories were challenged at t = 0 with 100 or 1,000 pfu of the initial virus dose of the SARS-CoV-2 2019-nCoV / USA-WA1 / 2020 strain (or the Delta or Omicron variant). The WA1 / 2020 strain was isolated from an oropharyngeal swab from a patient with respiratory disease who developed clinical illness (COVID-19) in January 2020 in Washington, US (BEI Resources, NR-52281). A virus dose of 10,000 pfu is considered to be significantly higher than the corresponding virus dose that a human subject would naturally encounter. Infected mice were dosed daily by forced oral gavage with (1) 0.5% sodium carboxymethylcellulose (CMC-Na) (negative control), (2) 100 and / or 300 mg / kg of Compound 2 in 0.5% CMC-Na (test groups), or (3) remdesivir (RDV). RDV, an antiviral agent that acts as a nucleoside analog and inhibits the RNA-dependent RNA polymerase (RdRp) of coronaviruses, including SARS-CoV-2, was used as a positive control. RDV was dissolved at 2.5 mg / mL in a vehicle containing 12% sulfobutyl ether-β-cyclodextrin sodium salt in water at pH 5.0 (containing HCl / NaOH) (25 mg / kg subcutaneous) and continued daily until the end of the study. Animals were monitored daily. Two animals were sacrificed at the indicated times (X-axis in Figures 13, 14, 15A, and 15B). The amount of virus in the lungs was determined by qPCR detecting viral RNA in lung lysates. Lung fibrosis was assayed using hematoxylin and eosin (H&E) staining (Figure 16).

[0185] As shown in FIGS. 13 and 14, treatment with Compound 2 resulted in a significant reduction in virus amount compared to both the negative control and the positive control. These data indicate that Compound 2 prevented the cellular entry of SARS-CoV-2 that requires the actin cytoskeleton. And as shown in FIGS. 15A and 15B, the effect was observed against both the Delta (FIG. 15A) and Omicron (FIG. 15B) variants of SARS-CoV-2. Furthermore, as shown in FIG. 16, treatment with Compound 2 had an additional benefit of reducing pulmonary fibrosis, which may help prevent long COVID. Example 3 Clinical treatment of COVID-19 using a ROCK2 inhibitor

[0186] A 7-day clinical trial was conducted to evaluate the efficacy of Compound 2 in the treatment of patients with mild to moderate COVID-19. After receiving a positive test result for the SARS-CoV-2 virus, participants were selected for the trial and received their first dose of placebo or Compound 2 (200 mg or 400 mg) within less than 72 hours after their first positive test. In total, 90 patients participated in the trial and were divided into three equally sized test groups (i.e., 30 patients / group): control, experimental group A, and experimental group B. Patients in the control group received standard treatment for mild to moderate COVID-19 and placebo once daily for 7 days. Patients in experimental group A received standard treatment for mild to moderate COVID-19 and Compound 2 (200 mg) once daily for 7 days. Patients in experimental group B received standard treatment for mild to moderate COVID-19 and Compound 2 (400 mg) once daily for 7 days. The virus amount in patients was measured daily to evaluate the time to virus clearance, clinical symptoms were observed and scored accordingly to determine the percentage of participants with resolved clinical symptoms. The results of the clinical trial are provided in FIGS. 17-20, which show the clinical efficacy of Compound 2 in the treatment of mild to moderate COVID-19.

[0187] Specifically, FIG. 17 shows that treatment with 400 mg of Compound 2 once daily (short dashed line - Experimental Group B) resulted in a substantial improvement in the sustained clinical symptom remission rate compared to treatment regimens including only standard treatment (solid line (control)) or standard treatment combined with daily administration of lower doses of Compound 2 (long dashed line - Experimental Group A). FIG. 18 shows that treatment with 200 mg (long dashed line - Experimental Group A) or 400 mg (short dashed line - Experimental Group B) of Compound 2 once daily resulted in a lower viral load in patients as measured by the negative conversion rate of SARS-CoV-2 nucleic acid detection compared to the control (solid line). FIGS. 19 and 20 show that treatment with Compound 2 (circles, long dashed line (Experimental Group A - 200 mg / day)) (squares, short dashed line (Experimental Group B - 400 mg / day)) reduced the level of inflammation in patients as measured by quantifying the levels of interleukin 6 (IL-6) and C-reactive protein (CRP), respectively. Example 4 Treatment of COVID-19 with ROCK2 Inhibitors

[0188] The test is conducted in men and women suspected of having or having SARS-CoV-2 infection. The test uses (2-(3-(4-((1H-indazol-5-yl)amino)quinazolin-2-yl)phenoxy)-N-isopropylacetamide) (Compound 1) and orally administers it at 100 - 400 mg daily for a treatment period of 2 - 3 weeks. Different amounts of Compound 1 are administered at specified intervals.

[0189] The subject is observed for about 2 weeks to about 6 months. Coronavirus testing is performed daily, every 2 days, every 3 days, or at other intervals during the treatment by collecting samples from the patient using a nasopharyngeal swab, an oropharyngeal swab, a nasal middle turbinate swab, or an anterior nares swab. Then, unless a point-of-care test for coronavirus is used, the swab is immediately placed into a sterile transport tube containing either about 2 - 3 mL of viral transport medium (VTM), Amies transport medium, or sterile saline. The samples are tested using CDC-approved testing methods. Different doses of Compound 1 administered at 100 mg / day, 200 mg / day, 300 mg / day, or 400 mg / day are also tested in patients according to the same protocol, and the dose-response relationship for the degree of virus inhibition is recorded. Example 5 Treatment of COVID-19 Using a ROCK2 Inhibitor and Remdesivir

[0190] The trial is conducted on male and female subjects suspected of having or having SARS-CoV-2 infection. The trial uses Compound 1, which is orally administered at 100 - 400 mg daily for a treatment period of 2 - 3 weeks. In parallel, the following schedule for adult and pediatric patients weighing at least 40 kg: on day 1, a loading dose of 200 mg; on days 2 - 10, remdesivir is administered by intravenous infusion in a total volume of up to 250 mL of 0.9% saline over 30 - 120 minutes according to a once-daily dose of about 100 mg for adult patients.

[0191] The subject is observed for about 2 weeks to about 6 months. Coronavirus testing is performed daily, every 2 days, every 3 days, or at other intervals during the treatment by collecting samples from the patient using a nasopharyngeal swab, an oropharyngeal swab, a nasal middle turbinate swab, or an anterior naris swab. Then, unless a point-of-care test for coronavirus is used, the swab is immediately placed into a sterile transport tube containing either about 2 - 3 mL of viral transport medium (VTM), Amies transport medium, or sterile saline. The samples are tested using CDC-approved testing methods. Different doses of Compound 1 administered at 100 mg / day, 200 mg / day, 300 mg / day, and 400 mg / day are also tested in patients according to the same protocol, and the dose-response relationship regarding the degree of virus inhibition is recorded. Example 6 Treatment of fibrosis using a ROCK2 inhibitor

[0192] Trials are conducted on men and women suspected of having or having SARS-CoV-2 infection and suspected of having or having fibrosis, in the same manner as described in Example 1. In addition to examining the degree of viral infection, respiratory symptoms, pulmonary function tests (PFT), and / or high-resolution computed tomography (HRCT) are performed on the patients at selected intervals, such as daily, every 2 days, every 3 days, every 4 days, or every 5 days, weekly, every 2 weeks, or monthly, etc., to monitor the progression of fibrosis. If clinically approved, X-ray examinations and / or CT scans may be performed. Further, the period during which Compound 1 is administered may be extended in this trial beyond 2 weeks at various dose levels.

[0193] To evaluate the lung function of each subject after treatment with Compound 1, the subjects are evaluated using a forced vital capacity (FVC) test. The forced expiratory volume in 1 second (FEV1), which is the amount of air forcefully exhaled in the first second of the FVC test, is measured for each subject on either the 1st, 3rd, 7th, 14th day of treatment, monthly, or every 2 months. Example 7 Treatment of fibrosis using a ROCK2 inhibitor and remdesivir

[0194] As in Example 2, conduct a study of 50 men and women suspected of having or having SARS-CoV-2 infection and suspected of having or having fibrosis. In addition to examining the degree of viral infection, respiratory symptoms, pulmonary function tests (PFTs), and / or high-resolution computed tomography (HRCT) are performed on the patients at selected intervals, such as daily, every 2 days, every 3 days, every 4 days, or every 5 days, weekly, every 2 weeks, or monthly, etc., to monitor the progression of fibrosis. If clinically approved, X-ray examinations and / or CT scans may be performed. Further, the period during which Compound 1 is administered may be extended in this study beyond 2 weeks at various dosage levels.

[0195] To evaluate the lung function of each subject after treatment with Compound 1, the subjects are evaluated using a forced vital capacity (FVC) test. The forced expiratory volume in 1 second (FEV1), which is the amount of air forcefully exhaled during the first second of the FVC test, is measured for each subject on day 1, day 3, day 7, day 14, monthly, or every 2 months of the treatment. Example 8 Treatment of Kawasaki disease-like diseases using a ROCK2 inhibitor

[0196] Conduct a study on pediatric patients suspected of having or having SARS-CoV-2 infection and suspected of having or having Kawasaki disease-like diseases, or showing pediatric multisystem inflammatory syndrome (PMIS) or pediatric inflammatory multisystem syndrome temporally associated with SARS-CoV-2 (PIMS-TS). The study uses Compound 1 and orally administers it at 100 mg / day, 200 mg / day, 300 mg / day, and 400 mg / day. Various amounts of Compound 1 are administered at specified intervals.

[0197] The subject is observed for about 2 weeks to about 6 months. Coronavirus testing is performed daily, every 2 days, every 3 days, or at other intervals during the treatment by collecting samples from the patient using a nasopharyngeal swab, an oropharyngeal swab, a nasal middle turbinate swab, or an anterior nares swab. Then, unless a point-of-care test for coronavirus is used, the swab is immediately placed into a sterile transport tube containing either about 2 - 3 mL of viral transport medium (VTM), Amies transport medium, or sterile saline. The samples are tested using CDC-approved testing methods. Different dosages of Compound 1 administered at 50 mg / day, 100 mg / day, 150 mg / day, 200 mg / day, 250 mg / day, 300 mg / day, 350 mg / day, 400 mg / day, 450 mg / day, 500 mg / day, 550 mg / day, 600 mg / day, 650 mg / day, 700 mg / day, 750 mg / day, or 800 mg / day are also tested in patients according to the same protocol, and the dose-response relationship is recorded for the degree of virus inhibition.

[0198] In addition to testing the degree of viral infection, for example, vasculitis tests such as blood tests (for C-reactive protein, complete blood count, amount of antineutrophil cytoplasmic antibody, or other biomarkers), urine tests (for red blood cell or protein content), and / or imaging tests (e.g., X-ray, ultrasound, computed tomography (CT), magnetic resonance imaging (MRI), and positron emission tomography (PET)) are performed on the patient at selected intervals such as daily, every 2 days, every 3 days, every 4 days, or every 5 days, weekly, every 2 weeks, or monthly to monitor the progression of Kawasaki-like disease, PMIS, or PIMS-TS. Further, the period during which Compound 1 is administered may be extended in this study beyond 2 weeks at various dosage levels.

[0199] To evaluate the progression of the disease, a two-dimensional echocardiogram is performed on the patient to assess coronary artery lesions (CAL) at 1 month of the illness. Measurements for each patient include the diameters of the proximal and middle portions of the left main coronary artery (LMCA), left anterior descending artery (LAD), left circumflex artery (LCX), and right coronary artery (RCA). The Z-score for each coronary artery is calculated. See Journal of the American Society of Echocardiography, 2011, 24(1). CAL can be defined as a z of at least 2 for any coronary artery of the LMCA, LAD, LCX, and proximal and middle portions of the RCA. Example 9 Treatment of Kawasaki disease-like disease using a ROCK2 inhibitor and remdesivir

[0200] Conduct a study of 20 pediatric patients suspected of having or having SARS-CoV-2 infection and suspected of having or having Kawasaki disease-like disease, or showing pediatric multisystem inflammatory syndrome (PMIS) or pediatric inflammatory multisystem syndrome temporally associated with SARS-CoV-2 (PIMS-TS). Administer Compound 1 orally at 100 - 400 mg daily for 2 - 3 weeks of treatment. Administer a predetermined amount of Compound 1 at specified intervals. In parallel, for pediatric patients with a body weight between 3.5 kg and 40 kg, a single loading dose of 5 mg / kg on Day 1; and remdesivir is administered by intravenous infusion over 30 - 120 minutes in a total volume of up to 125 mL of 0.9% saline at a daily loading dose of 2.5 mg / kg on Days 2 - 10.

[0201] The subject is observed for about 2 weeks to about 6 months. Coronavirus testing is performed daily, every 2 days, every 3 days, or at other intervals during the treatment by collecting samples from the patient using a nasopharyngeal swab, an oropharyngeal swab, a nasal middle turbinate swab, or an anterior naris swab. Then, unless a point-of-care test for coronavirus is used, the swab is immediately placed into a sterile transport tube containing either about 2 - 3 mL of viral transport medium (VTM), Amies transport medium, or sterile saline. The samples are tested using CDC-approved test methods. Different doses of Compound 1 administered at 50 mg / day, 100 mg / day, 150 mg / day, 200 mg / day, 250 mg / day, 300 mg / day, 350 mg / day, 400 mg / day, 450 mg / day, 500 mg / day, 550 mg / day, 600 mg / day, 650 mg / day, 700 mg / day, 750 mg / day, or 800 mg / day are also tested in patients according to the same protocol, and the dose-response relationship is recorded for the degree of virus inhibition.

[0202] In addition to testing the degree of viral infection, for example, perform vasculitis tests such as blood tests (for C-reactive protein, complete blood count, amount of antineutrophil cytoplasmic antibody, or other biomarkers), urine tests (for red blood cell or protein content), and / or imaging tests (e.g., X-ray, ultrasound, computed tomography (CT), magnetic resonance imaging (MRI), and positron emission tomography (PET)) on the patient at selected intervals such as daily, every two days, every three days, every four days, or every five days, weekly, every two weeks, or monthly to monitor the progression of Kawasaki-like disease, PMIS, or PIMS-TS. Further, the period during which Compound 1 is administered may be extended in this study beyond two weeks at various dosage levels. To evaluate the progression of the disease, perform two-dimensional echocardiography on the patient to evaluate coronary artery lesions (CAL) at one month of the disease. Measurements for each patient include the diameters of the proximal and middle portions of the left main coronary artery (LMCA), left anterior descending branch (LAD), left circumflex coronary artery (LCX), and right coronary artery (RCA). Calculate the Z-score for each coronary artery. See Renee Margossian et al., Journal of the American Society of Echocardiography, 2011, 24(1): 53-59. CAL can be defined as having a z of at least 2 in any of the coronary arteries of the LMCA, LAD, LCX, and the proximal and middle portions of the RCA.

Claims

1. A method for treating viral infection in a subject in need of treatment for viral infection, the method comprising administering to the subject a therapeutically effective amount of a ROCK2 inhibitor.

2. A method for preventing viral infection in a subject at risk of viral infection, the method comprising administering to the subject a prophylactically effective amount of a ROCK2 inhibitor.

3. A method for treating Kawasaki disease, PMIS, or PIMS-TS associated with viral infection in a subject in need of treatment for Kawasaki disease, PMIS, or PIMS-TS associated with viral infection, the method comprising administering to the patient a therapeutically effective amount of a ROCK2 inhibitor.

4. A method for treating or preventing sequelae resulting from viral infection in a subject in need of treatment or prevention of sequelae resulting from viral infection, the method comprising administering to the patient a therapeutically effective amount of a ROCK2 inhibitor.

5. The method according to any one of claims 1 to 4, wherein the ROCK2 inhibitor is a ROCK2-selective inhibitor.

6. The method according to any one of claims 1 to 5, wherein the ROCK2 inhibitor is ribonucleic acid (RNA).

7. The method according to claim 6, wherein the ROCK2 inhibitor is antisense RNA, small interfering RNA or microRNA.

8. The ROCK2 inhibitor is a compound having the structure of formula I: 【Chemical 24】 or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotopically labeled compound thereof, wherein R １ is selected from the group consisting of -O-(CH ２ ), ｒ CO ２ R １２ , -O-(CH ２ ), ｙ , -C(=O)NR １３ R １４ , -O-(CH ２ ), ｙ , -heteroaryl, -O-(CH ２ ), ｙ , -cycloalkyl, -O-C(=O)-(CH ２ ), ｙ , -NR １３ R １４ , -O-(CH ２ ), ｚ -NR １３ R １４ , -NH-C(=O)-(CH ２ ), ｙ , -NR １３ R １４ , -NH-C(=O)-X-R １５ , and -NH-(CH ２ ), ｙ , -NR １３ R １４ ; R １２ is C １ to C ６ alkyl, -(C １ to C ６ alkyl)-O-(C １ to C ６ alkyl), -(C １ to C ６ alkyl)-NR １６ R １７ , -(C １ to C ６ alkyl)-C(=O)NR １６ R １７ , -(C １ to C ６ alkyl)-O-(C １ to C ６ alkyl)-O-(C １ to C ６ alkyl), aryl, aralkyl, heteroaryl, C ３ to C ７ cycloalkyl, selected from the group consisting of 3- to 10-membered heterocyclic rings containing up to 3 heteroatoms, which may each be optionally substituted at one or more carbon atoms by 1 to 3 substituents independently selected from halo, C １ to C ６ alkoxy, hydroxy, amino, cyano and C １ to C ３ perfluoroalkyl; R １３ and R １４ are each independently selected from the group consisting of H, C １ ~C ８ alkyl, C ２ ~C ８ alkenyl, C ２ ~C ８ alkynyl, -(C １ ~C ６ alkyl)-O-(C １ ~C ６ alkyl), -(C １ ~C ６ alkyl)-NR １６ R １７ , -(C １ ~C ６ alkyl)-C(=O)NR １６ R １７ , aryl, aralkyl, heteroaryl, C ３ ~C ７ cycloalkyl, and are each independently selected from the group consisting of 3- to 10-membered heterocyclic rings containing up to 3 heteroatoms, and each of them is optionally substituted by 1 to 3 substituents independently selected from halo, C １ ~C ６ alkyl, C ２ ~C ６ alkenyl, C ３ ~C ７ cycloalkyl, C １ ~C ６ alkoxy, hydroxy, amino, cyano and C １ ~C ３ perfluoroalkyl; or R １３ and R １４ together form a halo, C １ to C ６ alkyl, C ２ to C ６ alkenyl, C １ to C ６ alkoxy, C ３ to C ７ cycloalkyl, oxo, hydroxy, amino, cyano and C １ to C ３ optionally substituted by 1 to 3 substituents independently selected from perfluoroalkyl, and may form a 3- to 10-membered heterocyclic ring having up to 3 heteroatoms; X is absent or is selected from -O, NH, and C １ ~C ６ alkyl; R １５ is selected from the group consisting of heteroaryl, C ３ -C ７ -cycloalkyl, and 3- to 10-membered heterocyclic rings containing up to 3 heteroatoms, each of which is independently selected from halo, C １ -C ６ -alkyl, C ２ -C ６ -alkenyl, C １ -C ６ -alkoxy, hydroxy, amino, cyano and C １ -C ３ -perfluoroalkyl, and may be optionally substituted by 1 to 3 substituents independently selected therefrom; or R １５ is -(C １ ~C ６ alkyl)-O-(C １ ~C ６ alkyl), -(C １ ~C ６ alkyl)-NR １６ R １７ , -CO ２ R １８ , -O-(CH ２ ), ｘ -CO ２ R １８ , and -C(=O)NR １６ R １７ is selected from; R １６ and R １７ are each independently selected from the group consisting of H, C １ to C ８ alkyl, C ２ to C ８ alkenyl, C ２ to C ８ alkynyl, -(C １ to C ６ alkyl)-O-(C １ to C ６ alkyl), aryl, aralkyl, heteroaryl, C ３ to C ７ cycloalkyl, and 3- to 10-membered heterocyclic rings containing up to 3 heteroatoms, and each of them may optionally be substituted by 1 to 3 substituents independently selected from halo, C １ to C ６ alkyl, C ２ to C ６ alkenyl, C １ to C ６ alkoxy, hydroxy, amino, cyano and C １ to C ３ perfluoroalkyl; or R １６ and R １７ together form a halo, C １ -C ６ alkyl, C ２ -C ６ alkenyl, C １ -C ６ alkoxy, oxo, hydroxy, amino, cyano and C １ -C ３ optionally substituted by 1 to 3 substituents independently selected from perfluoroalkyl, and may form a 3- to 10-membered heterocyclic ring having up to 3 heteroatoms; R １８ is selected from the group consisting of H, aryl, aralkyl, heteroaryl, C １ to C ６ alkyl, -(C １ to C ６ alkyl)-O-(C １ to C ６ alkyl), -(C １ to C ６ alkyl)-NR １６ R １７ , -(C １ to C ６ alkyl)-O-(C １ to C ６ alkyl)-O-(C １ to C ６ alkyl), and may be optionally substituted by 1 to 3 substituents independently selected from halo, C １ to C ６ alkoxy, hydroxy, amino, cyano and C １ to C ３ perfluoroalkyl; x is selected from 0 to 6; y is selected from 0 to 6; z is selected from 2 to 6; Each R ２ is independently selected from the group consisting of lower alkyl, CN, halo, hydroxy, lower alkoxy, amino, and perfluoro lower alkyl; Each R ３ is independently selected from the group consisting of lower alkyl, CN, halo, hydroxy, lower alkoxy, amino, and perfluoro lower alkyl; R ４ is selected from H, -(CH ２ ) ａ -NR ４３ R ４４ , -Y-R ４２ , -O-(CH ２ ) ａ -COR ２ R ４２ , -O-(CH ２ ) ａ -C(=O)NR ４３ R ４４ , -O-(CH ２ ) ａ -heteroaryl, -O-(CH ２ ) ａ -cycloalkyl, -O-C(=O)-(CH ２ ) ａ -NR ４３ R ４４ , -O-(CH ２ ) ｃ -N ４３ R ４４ , -NH-C(=O)-Y-R ４５ , -NH-C(=O)-(CH ２ ) ａ -NR ４３ R ４４ ; R ４２ is selected from the group consisting of C １ to C ６ alkyl, -(C １ to C ６ alkyl)-O-(C １ to C ６ alkyl), -(C １ to C ６ alkyl)-NR ４６ R ４７ , -(C １ to C ６ alkyl)-C(=O)NR ４６ R ４７ , -(C １ to C ６ alkyl)-O-(C １ to C ６ alkyl)-O-(C １ to C ６ alkyl), and may be optionally substituted at one or more carbon atoms by one to three substituents independently selected from halo, C １ to C ６ alkoxy, hydroxy, amino, cyano, and C １ to C ３ perfluoroalkyl; R ４３ and R ４４ are each independently selected from the group consisting of H, C １ ~C ８ alkyl, C ２ ~C ８ alkenyl, C ２ ~C ８ alkynyl, -(C １ ~C ６ alkyl)-O-(C １ ~C ６ alkyl), -(C １ ~C ６ alkyl)-NR ４６ R ４７ , -(C １ ~C ６ alkyl)-C(=O)NR ４６ R ４７ , aryl, aralkyl, heteroaryl, C ３ ~C ７ cycloalkyl, and are each independently selected from the group consisting of 3- to 10-membered heterocyclic rings containing up to 3 heteroatoms, and they are each optionally substituted by 1 to 3 substituents independently selected from halo, C １ ~C ６ alkyl, C ２ ~C ６ alkenyl, C ３ ~C ７ cycloalkyl, C １ ~C ６ alkoxy, hydroxy, amino, cyano and C １ ~C ３ perfluoroalkyl; or R ４３ and R ４４ together form a halo, C １ to C ６ alkyl, C ２ to C ６ alkenyl, C １ to C ６ alkoxy, oxo, hydroxy, amino, cyano and C １ to C ３ optionally substituted by 1 to 3 substituents independently selected from perfluoroalkyl, and may form a 3- to 10-membered heterocyclic ring having up to 3 heteroatoms; Y is absent or is selected from O, NH, and C １ -C ６ -alkyl; R ４５ is selected from the group consisting of H, aryl, -(C １ ~C ６ alkyl)-O-(C １ ~C ６ alkyl), -(C １ ~C ６ alkyl)-NR ４６ R ４７ , -CO ２ R ４８ , -O-(CH ２ ) ６ -CO ２ R ４８ , and -C(=O)NR ４６ R ４７ ; R ４６ and R ４７ are each independently selected from the group consisting of H, C １ ~C ８ alkyl, C ２ ~C ８ alkenyl, C ２ ~C ８ alkynyl, -(C １ ~C ６ alkyl)-O-(C １ ~C ６ alkyl), aryl, aralkyl, heteroaryl, C ３ ~C ７ cycloalkyl, and 3- to 10-membered heterocyclic rings containing up to 3 heteroatoms, and are each optionally substituted by 1 to 3 substituents independently selected from halo, C １ ~C ６ alkyl, C ２ ~C ６ alkenyl, C １ ~C ６ alkoxy, hydroxy, amino, cyano and C １ ~C ３ perfluoroalkyl; or R ４６ and R ４７ together form a halo, C １ -C ６ alkyl, C ２ -C ６ alkenyl, C １ -C ６ alkoxy, oxo, hydroxy, amino, cyano and C １ -C ３ optionally substituted by 1 to 3 substituents independently selected from perfluoroalkyl and having a 3- to 10-membered heterocyclic ring having up to 3 heteroatoms; R ４８ is selected from the group consisting of H, aryl, aralkyl, heteroaryl, C １ to C ６ alkyl, -(C １ to C ６ alkyl)-O-(C １ to C ６ alkyl), -(C １ to C ６ alkyl)-NR ４６ R ４７ , -(C １ to C ６ alkyl)-O-(C １ to C ６ alkyl)-O-(C １ to C ６ alkyl), and may be optionally substituted by 1 to 3 substituents independently selected from halo, C １ to C ６ alkoxy, hydroxy, amino, cyano, and C １ to C ３ perfluoroalkyl; a is selected from 0 to 6; b is selected from 0 to 6; c is selected from 2 to 6; R ５ is selected from the group consisting of H, C １ ~C ６ alkyl, -(CH ２ ) ｄ -C(=O)-NR ５３ R ５４ , -C(=O)-(CH ２ ) ｄ -NR ５３ R ５４ , and -C(=O)-X-R ５５ ; R ５３ and R ５４ are each independently selected from the group consisting of H, C １ ~C ８ alkyl, C ２ ~C ８ alkenyl, C ２ ~C ８ alkynyl, -(C １ ~C ６ alkyl)-O-(C １ ~C ６ alkyl), -(C １ ~C ６ alkyl)-NR ５６ R ５７ , -(C １ ~C ６ alkyl)-C(=O)NR ５６ R ５７ , aryl, aralkyl, heteroaryl, C ３ ~C ７ cycloalkyl, and are each independently selected from the group consisting of 3- to 10-membered heterocyclic rings containing up to 3 heteroatoms, and each of them is optionally substituted by 1 to 3 substituents independently selected from halo, C １ ~C ６ alkyl, C ２ ~C ６ alkenyl, C ３ ~C ７ cycloalkyl, C １ ~C ６ alkoxy, hydroxy, amino, cyano and C １ ~C ３ perfluoroalkyl; or R ５３ and R ５４ together form a halo, C １ -C ６ alkyl, C ２ -C ６ alkenyl, C １ -C ６ alkoxy, C ３ -C ７ cycloalkyl, oxo, hydroxy, amino, cyano and C １ -C ３ optionally substituted by 1 to 3 substituents independently selected from perfluoroalkyl, and may form a 3- to 10-membered heterocyclic ring having up to 3 heteroatoms; R ５５ is selected from the group consisting of H, aryl, -(C １ ~C ６ alkyl)-O-(C １ ~C ６ alkyl), -(C １ ~C ６ alkyl)-NR ５６ R ５７ , -CO ２ R ５８ , -O-(CH ２ ) ｅ , -CO ２ R ５８ , and -C(=O)NR ５６ R ５７ ; R ５６ and R ５７ are each independently selected from the group consisting of H, C １ to C ８ alkyl, C ２ to C ８ alkenyl, C ２ to C ８ alkynyl, -(C １ to C ６ alkyl)-O-(C １ to C ６ alkyl), aryl, aralkyl, heteroaryl, C ３ to C ７ cycloalkyl, and 3- to 10-membered heterocyclic rings containing up to 3 heteroatoms, and each of them is optionally substituted by 1 to 3 substituents independently selected from halo, C １ to C ６ alkyl, C ２ to C ６ alkenyl, C １ to C ６ alkoxy, hydroxy, amino, cyano and C １ to C ３ perfluoroalkyl; or R ５６ and R ５７ together form a halo, C １ -C ６ alkyl, C ２ -C ６ alkenyl, C １ -C ６ alkoxy, oxo, hydroxy, amino, cyano and C １ -C ３ optionally substituted by 1 to 3 substituents independently selected from perfluoroalkyl, and may form a 3- to 10-membered heterocyclic ring having up to 3 heteroatoms; R ５８ is selected from the group consisting of H, aryl, aralkyl, heteroaryl, C １ to C ６ alkyl, -(C １ to C ６ alkyl)-O-(C １ to C ６ alkyl), -(C １ to C ６ alkyl)-NR ５６ R ５７ , -(C １ to C ６ alkyl)-O-(C １ to C ６ alkyl)-O-(C １ to C ６ alkyl), and may be optionally substituted by 1 to 3 substituents independently selected from halo, C １ to C ６ alkoxy, hydroxy, amino, cyano and C １ to C ３ perfluoroalkyl; d is selected from 0 to 6; e is selected from 0 to 6; R ６ is selected from the group consisting of H, C １ ~C ６ alkyl, -(CH ２ ) ｒ -C(=O)-NR ６３ R ６４ , -C(=O)-(CH ２ ) ｒ -NR ６３ R ６４ , and -C(=O)-X-R ６５ ; R ６３ and R ６４ are each independently selected from the group consisting of H, C １ ~C ８ alkyl, C ２ ~C ８ alkenyl, C ２ ~C ８ alkynyl, -(C １ ~C ６ alkyl)-O-(C １ ~C ６ alkyl), -(C １ ~C ６ alkyl)-NR ６６ R ６７ , -(C １ ~C ６ alkyl)-C(=O)NR ６６ R ６７ , aryl, aralkyl, heteroaryl, C ３ ~C ７ cycloalkyl, and are each independently selected from the group consisting of 3- to 10-membered heterocyclic rings containing up to 3 heteroatoms, and each of them is optionally substituted with 1 to 3 substituents independently selected from halo, C １ ~C ６ alkyl, C ２ ~C ６ alkenyl, C ３ ~C ７ cycloalkyl, C １ ~C ６ alkoxy, hydroxy, amino, cyano and C １ ~C ３ perfluoroalkyl; or R ６３ and R ６４ together form a 3- to 10-membered heterocyclic ring having up to 3 heteroatoms, optionally substituted by 1 to 3 substituents independently selected from halo, C １ to C ６ alkyl, C ２ to C ６ alkenyl, C １ to C ６ alkoxy, C ３ to C ７ cycloalkyl, oxo, hydroxy, amino, cyano and C １ to C ３ perfluoroalkyl; R ６５ is selected from the group consisting of H, aryl, -(C １ ~C ６ alkyl)-O-(C １ ~C ６ alkyl), -(C １ ~C ６ alkyl)-NR ６６ R ６７ , -CO ２ R, -O-(CH ２ ), -CO ２ R ６８ , and -C(=O)NR ６６ R ６７ ; R ６６ and R ６７ are each independently selected from the group consisting of H, C １ to C ５ alkyl, C ２ to C ８ alkenyl, C ２ to C ８ alkynyl, -(C １ to C ６ alkyl)-O-(C １ to C ６ alkyl), aryl, aralkyl, heteroaryl, C ３ to C ７ cycloalkyl, a 3- to 10-membered heterocyclic ring containing up to 3 heteroatoms, and are each optionally substituted by 1 to 3 substituents independently selected from halo, C １ to C ６ alkyl, C ２ to C ６ alkenyl, C １ to C ６ alkoxy, hydroxy, amino, cyano and C １ to C ３ perfluoroalkyl; or R ６６ and R ６７ together form a halo, C １ -C ６ alkyl, C ２ -C ６ alkenyl, C １ -C ６ alkoxy, oxo, hydroxy, amino, cyano and C １ -C ３ optionally substituted by 1 to 3 substituents independently selected from perfluoroalkyl, and may form a 3- to 10-membered heterocyclic ring having up to 3 heteroatoms; R ６８ is selected from the group consisting of H, aryl, aralkyl, heteroaryl, C １ to C ６ alkyl, -(C １ to C ６ alkyl)-O-(C １ to C ６ alkyl), -(C １ to C ６ alkyl)-NR ６６ R ６７ , -(C １ to C ６ alkyl)-O-(C １ to C ６ alkyl)-O-(C １ to C ６ alkyl), and may be optionally substituted by 1 to 3 substituents independently selected from halo, C １ to C ６ alkoxy, hydroxy, amino, cyano and C １ to C ３ perfluoroalkyl; r is selected from 0 to 6; s is selected from 0 to 6; n is selected from 0 to 4; m is selected from 0 to 3; p is selected from 0 and 1, the method according to any one of claims 1 to 5.

9. The ROCK2 inhibitor has the structure of formula II: 【Chemical 25】 A compound having or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotopically labeled compound thereof, wherein R １ , R ２ , R ３ , R ５ , R ６ , m and n are as defined in claim 8, the method according to claim 8.

10. The ROCK2 inhibitor has the structure of formula III: 【Chemical 26】 or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotopically labeled compound thereof, R １３ and R １４ are each independently selected from the group consisting of H, C １ ~C ８ alkyl, C ２ ~C ８ alkenyl, C ２ ~C ８ alkynyl, -(C １ ~C ６ alkyl)-O-(C １ ~C ６ alkyl), -(C １ ~C ６ alkyl)-NR １６ R １７ , -(C １ ~C ６ alkyl)-C(=O)NR １６ R １７ , aryl, aralkyl, heteroaryl, C ３ ~C ７ cycloalkyl, and a 3- to 10-membered heterocyclic ring containing up to 3 heteroatoms, and are each optionally substituted by 1 to 3 substituents independently selected from halo, C １ ~C ６ alkyl, C ２ ~C ６ alkenyl, C ３ ~C ７ cycloalkyl, C １ ~C ６ alkoxy, hydroxy, amino, cyano and C １ ~C ３ perfluoroalkyl; Or R １３ And R １４ Together may form a 3- to 10-membered heterocyclic ring having up to 3 heteroatoms optionally substituted by 1 to 3 substituents independently selected from halo, C １ To C ６ Alkyl, C ２ To C ６ Alkenyl, C １ To C ６ Alkoxy, C ３ To C ７ Cycloalkyl, oxo, hydroxy, amino, cyano and C １ To C ３ Perfluoroalkyl; R １６ and R １７ are each independently selected from the group consisting of H, C １ to C ８ alkyl, C ２ to C ８ alkenyl, C ２ to C ８ alkynyl, -(C １ to C ６ alkyl)-O-(C １ to C ６ alkyl), aryl, aralkyl, heteroaryl, C ３ to C ７ cycloalkyl, and 3- to 10-membered heterocyclic rings containing up to 3 heteroatoms, and each of them may optionally be substituted by 1 to 3 substituents independently selected from halo, C １ to C ６ alkyl, C ２ to C ６ alkenyl, C １ to C ６ alkoxy, hydroxy, amino, cyano and C １ to C ３ perfluoroalkyl; or R １６ and R １７ together form a halo, C １ -C ６ alkyl, C ２ -C ６ alkenyl, C １ -C ６ alkoxy, oxo, hydroxy, amino, cyano and C １ -C ３ optionally substituted by 1 to 3 substituents independently selected from perfluoroalkyl, and may form a 3- to 10-membered heterocyclic ring having up to 3 heteroatoms; R ２ 、R ３ 、R ５ 、R ６ 、m and n are each as defined in claim 8, the method according to claim 9.

11. The ROCK2 inhibitor has the structure of formula IV: 【Chemical 27】 A compound having the same or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotopically labeled compound thereof, wherein R １３ and R １４ are as defined in claim 10, the method according to claim 10.

12. The ROCK2 inhibitor is a compound having the chemical structure of Compound 1 (2-(3-(4-((1H-indazol-5-yl)amino)quinazolin-2-yl)phenoxy)-N-isopropylacetamide): 【Chemical 28】 Or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotopically labeled compound thereof, the method according to claim 11.

13. The method according to any one of claims 1 to 12, wherein the viral infection is caused by a coronavirus.

14. The method according to any one of claims 1 to 13, wherein the viral infection is caused by SARS-CoV-1, SARS-CoV-2, or MERS-CoV.

15. The method according to any one of claims 1 to 14, wherein the viral infection is caused by SARS-CoV-2.

16. The method according to claim 15, wherein the viral infection is caused by the Delta or Omicron variant of SARS-CoV-2.

17. The sequelae resulting from the viral infection include one or more of the group consisting of fatigue, dyspnea, cough, arthralgia, myalgia, headache, chest pain, fever, palpitations, myocarditis, ventricular dysfunction, stroke, abnormal pulmonary function, pulmonary fibrosis, renal insufficiency, rash, alopecia, olfactory and / or gustatory dysfunction, sleep regulation disorder, cognitive function disorder change, memory function disorder, depression, anxiety, mood swings, and combinations thereof. The method according to any one of claims 4 to 16.

18. The method according to any one of claims 4 to 17, wherein the sequelae is pulmonary fibrosis.

19. The method according to any one of claims 1 to 18, further comprising administering to the subject a therapeutically effective amount of at least one other therapeutic agent.

20. The method according to claim 19, wherein the at least one other therapeutic agent is selected from the group consisting of antiviral agents, corticosteroids, anti-inflammatory signal transduction modulators, β2-adrenergic receptor agonists bronchodilators, anticholinergics, mucolytics, hypertonic saline, and combinations thereof.

21. The method according to claim 19, wherein the at least one other therapeutic agent comprises remdesivir.

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