ROCK2 inhibitors for the treatment of viral infections

Abstract

The present disclosure provides compositions and methods comprising selective inhibitors of Rho-associated coiled-coil kinase 2 (ROCK2) for use in the treatment of viral infections, particularly coronavirus infections such as SARS-CoV-2, and for the treatment of sequelae of viral infections, including sequelae of coronavirus infections.

Description

[Technical field]

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

[0002] Viral infections are common between animals and humans. For example, the COVID-19 pandemic caused by the SARS-CoV-2 virus has become a global public health threat. Alarmingly, the pandemic has infected over 100 million people worldwide and killed over 2 million people by the end of 2020.

[0003] Coronaviruses are a large family of enveloped viruses with a positive-sense single-stranded RNA genome and belong to the order Nidovirales and family Coronaviridae. Coronavirus infections are primarily concentrated in the upper respiratory system and gastrointestinal tract, but the lower respiratory system may be involved in more severe infections. Depending on the specific virus and host cell type, the symptoms and pathological damage caused by coronavirus infections may be quite different. Some coronaviruses, such as HCoV-NL63, HCoV-229E, and HCoV-0C43, circulate continuously in the human population and cause mild symptoms similar to the common cold. Other coronaviruses, such as 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 has spread to several other countries. SARS-CoV-1 was first identified in China in 2002 and caused a global pandemic in 2002 and 2003. Other human coronaviruses include 222E (alphacoronavirus), NL63 (alphacoronavirus), OC43 (betacoronavirus), and HKU1 (betacoronavirus).

[0004] Survivors of coronavirus infections, including those with Covid-19, have reported persistent and diverse postviral symptoms, even if the initial disease course was mild. Currently, there is no cure for postviral syndrome, and treatment is directed at alleviating and managing symptoms. Furthermore, the economic impact of postviral syndrome can be significant, including lost productivity and employment, and increased need for disability benefits and financial support. There is a high and unmet need for therapeutics to treat coronavirus infection and to treat the sequelae of coronavirus infection.

[0005] Rho-associated coiled-coil kinase (ROCK) is a serine / threonine kinase of the AGC (PKA, PKG, and PKC) kinase family, and is composed of two isoforms, ROCK1 and ROCK2. These two isoforms are differentially expressed and regulated in certain tissues. For example, ROCK1 is ubiquitously expressed at relatively high levels, whereas ROCK2 is selectively expressed in certain tissues, such as the heart, brain, and skeletal muscle. ROCK is a target of the small GTPase Rho and is involved in various cellular activities carried out by phosphorylating downstream effector proteins (e.g., MLC, LIMK, ERM, MARCKS, CRMP-2, etc.). Studies have shown that various diseases (e.g., pulmonary fibrosis, cardio-cerebrovascular disease, neurological diseases, and cancer, etc.) are associated with pathways mediated by ROCK. Therefore, ROCK is considered an important target in new drug development.

[0006] The present disclosure relates to surprisingly potent and previously unrecognized antiviral effects of ROCK2 inhibitors and their use in treating viral infections, including coronavirus infections such as SARS-CoV-1, SARS-CoV-2, or MERS-CoV infections. Additionally, ROCK2 inhibition treats many secondary symptoms that can result from viral infections, including inflammation, fibrosis, and cytokine storm. Summary of the Invention

[0007] In one aspect, the disclosure provides an antiviral composition comprising a Rho-associated coiled-coil kinase (ROCK2) inhibitor. In one aspect, the disclosure provides a method of treating a subject by administering to the subject a therapeutically effective amount of a ROCK2 inhibitor, or a composition comprising a ROCK2 inhibitor. Such methods and compositions described herein may be useful for treating an individual suffering from or suspected of suffering from a viral infection. In an embodiment, the viral infection is a coronavirus infection. In an embodiment, the viral infection is a SARS-CoV-1 infection, a SARS-CoV-2 infection, or a MERS-CoV infection. In an embodiment, the viral infection is caused by SARS-CoV-2 variant delta and / or SARS-CoV-2 variant omicron. In an embodiment, the methods and compositions described herein are useful for treating and preventing sequelae from a viral infection, including sequelae from a coronavirus infection. In an embodiment, the methods and compositions described herein are useful for treating and preventing sequelae from a SARS-CoV-1 infection, a SARS-CoV-2 infection, or a MERS-CoV infection. In embodiments, the viral infection is caused by SARS-CoV-2 variant delta and / or SARS-CoV-2 variant omicron. In embodiments, the sequelae include one or more of the group consisting of fatigue, dyspnea (difficulty breathing), cough, arthralgia (pain in the joints), muscle pain, headache, chest pain, fever, palpitations, myocardial inflammation, ventricular dysfunction, stroke, pulmonary function abnormalities, fibrosis (such as pulmonary fibrosis), renal dysfunction rash, alopecia, smell and / or taste disorders, sleep regulation disorders, cognitive impairment changes, memory impairment, depression, anxiety, mood changes, and combinations thereof. In embodiments, the sequelae include fibrosis. In one aspect, the disclosure provides a method of treating a viral infection in a subject in need thereof, comprising administering to the human subject a therapeutically effective amount of a ROCK2 selective inhibitor, wherein the ROCK2 selective inhibitor is a compound having formulas I-XII, in particular (6-(4-((4-(1H-pyrazol-4-yl)phenyl)amino)pyrimidin-2-yl)-1-methyl-1H-indol-2-yl)(3,3-difluoroazetidin-1-yl)methanone.In one aspect, the disclosure provides a method of treating and preventing sequelae of a viral infection in a subject in need thereof, comprising administering to the human subject a therapeutically effective amount of a ROCK2 selective inhibitor, the ROCK2 selective inhibitor being a compound having formula I-XII, in particular (6-(4-((4-(1H-pyrazol-4-yl)phenyl)amino)pyrimidin-2-yl)-1-methyl-1H-indol-2-yl)(3,3-difluoroazetidin-1-yl)methanone.

[0008] In some embodiments, the method includes administering to the subject a therapeutically effective amount of at least one other therapeutic agent, which can be another antiviral agent, a corticosteroid, an anti-inflammatory signaling modulator, a beta2 adrenergic receptor agonist bronchodilator, an anticholinergic agent, a mucolytic agent, hypertonic saline, or a mixture thereof.

[0009] In one aspect, the disclosure provides a method of treating a viral infection in a subject in need thereof, comprising administering to a human subject a therapeutically effective amount of a ROCK2 selective inhibitor, the ROCK2 selective inhibitor being a compound having Formulas I-XII, in particular (6-(4-((4-(1H-pyrazol-4-yl)phenyl)amino)pyrimidin-2-yl)-1-methyl-1H-indol-2-yl)(3,3-difluoroazetidin-1-yl)methanone, and a therapeutically effective amount of at least one other antiviral agent. The at least one other antiviral agent may be a nucleoside or nucleotide analog, or a pharma-ceutically acceptable salt or prodrug thereof.

[0010] In an embodiment, a method of treating a viral infection in a subject in need of such treatment or a method of treating and preventing sequelae of a viral infection in a subject in need of such treatment and prevention is provided, comprising administering to the patient a ROCK2 inhibitor disclosed herein at a total dose of about 200 mg to about 500 mg per day. In one embodiment, the ROCK2 inhibitor is administered to the patient at a total dose of about 200 mg per day. In one embodiment, the ROCK2 inhibitor is administered to the patient at a total dose of about 300 mg per day. In one embodiment, the ROCK2 inhibitor is administered to the patient at a total dose of about 400 mg per day. In one embodiment, the ROCK2 inhibitor is administered to the patient at a total dose of about 500 mg per day. In one embodiment, the ROCK2 inhibitor is administered in one daily dose. In one embodiment, the ROCK2 inhibitor is administered in two daily doses. In embodiments, the ROCK2 inhibitor is administered to the patient within about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 24, about 28, about 72, about 96, or about 120 hours after the patient is first exposed to the virus causing the viral infection. In embodiments, the ROCK2 inhibitor is administered to the patient within about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 24, about 28, about 72, about 96, or about 120 hours after the patient develops symptoms caused by the viral infection. In embodiments, the ROCK2 inhibitor is administered to the patient for about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, or about 10 weeks after the symptoms have subsided. [Brief description of the drawings]

[0011] [Figure 1] Cytotoxicity: LDH assay. Immortalized human airway epithelial cells (Calu-3) were treated with the compounds of formula XII at the indicated concentrations for 2 hours at 37° C., and then challenged with SARS-COV-2 for 6 hours. After washing, cells were maintained for 48 hours and LDH assay was performed. [Diagram 2]Efficacy: SARS-COV-2 CPN in the supernatant. Immortalized human airway epithelial cells (Calu-3) were treated with the compound of formula XII at the indicated concentrations for 2 hours at 37°C and then challenged with SARS-COV-2 for 6 hours. After washing, cells were maintained for 48 hours and viral infection was analyzed by qPCR of the supernatant. Each data point represents the average of triplicate replicates. [Diagram 3] Efficacy: Classical dose-response curve. The dose-response curve for the experiment described in Example 1 and Figure 2 is shown. Based on the best fit, the EC50 was calculated as 124 nM. [Figure 4] Efficacy: Median-effect model analysis. Median-effect model analysis for the experiments described in Example 1 and Figure 2. Fa = proportion affected, Fu = proportion unaffected, D = dose (μM). [Diagram 5] Efficacy: predicted values ​​from EC50 to EC97. Predicted values ​​from EC50 to EC97 in the experiments described in Example 1 and FIG. [Figure 6A] Efficacy: viral load in lung lysates. Formula XII reduces spread of SARS-CoV2 in mouse lungs in vivo. Viral copies per mg of lung at an initial viral load of 100 pfu of SARS-CoV2 are shown. Groups from left to right: vehicle, Formula XII (300 mg / kg), Remdesivir (RDV). Inset: Formula XII (300 mg / kg) (left), Remdesivir (RDV) (right). [Figure 6B] Efficacy: viral load in lung lysates. Formula XII reduces spread of SARS-CoV2 in mouse lungs in vivo. Viral copies per mg of lung at an initial viral load of 1000 pfu of SARS-CoV2 are shown. Groups from left to right: vehicle, Formula XII (300 mg / kg), remdesivir (RDV). Inset: Formula XII (300 mg / kg) (left), remdesivir (RDV) (right). [Figure 6C]Efficacy: viral load in lung lysates. Formula XII reduces spread of SARS-CoV2 in mouse lungs in vivo. Viral copies per mg of lung at an initial viral load of 10,000 pfu of SARS-CoV2 are shown. Groups from left to right: vehicle, Formula XII (300 mg / kg), remdesivir (RDV). Inset: Formula XII (300 mg / kg) (left), remdesivir (RDV) (right). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] The compounds, compositions and methods described herein provide selective inhibitors of Rho-associated coiled-coil kinase 2 (ROCK2) for use in the treatment of viral infections, particularly coronavirus infections such as SARS-CoV-1 infection, SARS-CoV-2 infection, or MERS-CoV infection, and in the treatment and prevention of sequelae of viral infections, including sequelae of coronavirus infection.

[0013] ROCK2 inhibitors The compounds used in the methods and compositions disclosed herein are ROCK inhibitors, particularly selective inhibitors of ROCK 2. These compounds provide excellent inhibitory activity of ROCK (preferably ROCK 2), good selectivity (higher selectivity for ROCK 2 compared to ROCK 1), good physicochemical properties (e.g., solubility, physical and / or chemical stability), improved pharmacokinetic properties (e.g., improved bioavailability, suitable half-life and duration of action), improved safety (low toxicity and / or fewer side effects, broad therapeutic range), and the like.

[0014] As provided herein, ROCK2 inhibitors have surprisingly potent antiviral effects that have not been previously recognized. Without being bound by theory, it is believed that ROCK2 inhibitors, such as compounds of formulas I-XII, interfere with one or more of the following: (1) the pathways that viruses use to enter cells; (2) the cytoskeleton that viruses use as a pathway for movement and spread; (3) the pathways that viruses use to upregulate cellular energy metabolism; and (4) the pathways that viruses use to spread to other cells. Inhibitors of ROCK2 interfere with the interaction of viruses with cytoskeletal actin filaments, microtubules, and / or intermediate filaments, which are deeply involved in the life cycle and pathological damage caused by viruses, particularly coronaviruses. In addition, ROCK2 inhibitors exert potent effects on many secondary symptoms caused by viral infections, such as inflammation, fibrosis, cytokines, and bradykinin storms.

[0015] According to one aspect, the disclosure provides a method of treating a subject by administering to the subject a therapeutically effective amount of a selective ROCK2 inhibitor, or a composition comprising a selective ROCK2 inhibitor, wherein the ROCK2 selective inhibitor has the structure of Formula I: [ka] or a pharma- ceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotopically labeled compound thereof, wherein X and Y each independently represent a direct bond, C(=O), O, or 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 3~10 Cyclic hydrocarbyl, saturated or partially unsaturated 3-10 membered heterocyclyl, C 6~10 Aryl, 5-14 membered heteroaryl and C 6~12aralkyl, wherein up to two ring members of the cyclic hydrocarbyl and heterocyclyl are C(=O); Ring A and ring B are each independently a saturated or partially unsaturated C 3~10 Hydrocarbon ring, saturated or partially unsaturated 3-10 membered heterocyclic ring, C 6~10 and 5-14 membered heteroaromatic rings, wherein up to two ring members of the hydrocarbon ring and the 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 via a nitrogen atom; Ring C is saturated or partially unsaturated C 3~10 Hydrocarbon ring, saturated or partially unsaturated 3-10 membered heterocyclic ring, C 6~10 selected from the group consisting of aromatic rings and 5-14 membered heteroaromatic rings, wherein up to two ring members of the hydrocarbon ring and the heterocyclic ring are C(=O); Ring D is absent or saturated or partially unsaturated C 3~10 Hydrocarbon ring, saturated or partially unsaturated 3-10 membered heterocyclic ring, C 6~10 selected from the group consisting of aromatic rings and 5-14 membered heteroaromatic rings, wherein up to two ring members of the hydrocarbon ring and the heterocyclic ring are C(=O); Ring E is selected from the group consisting of: [ka] Ring F is a saturated or partially unsaturated C 3~10 Hydrocarbon ring, saturated or partially unsaturated 3-10 membered heterocyclic ring, C 6~10 selected from the group consisting of aromatic rings and 5-14 membered heteroaromatic rings, wherein up to two ring members of the hydrocarbon ring and the heterocyclic ring are C(=O); R 1 is H, -NH2, C 1~6 Alkyl, C 6~10 Aryl, 5-14 membered heteroaryl, N-methylpyrrolidinyl, N-methylpiperidinyl, [ka] Acetyl, [ka] -C(=O)-(C 1~6 alkylene)n-CF3, -C(=O)-(C 1~6 Alkylene)CN, -C(=O)-(saturated or partially unsaturated C 3~10 Cyclic hydrocarbyl), -NHC(=O)-(saturated or partially unsaturated C 3~10 -C(=O)-(saturated or partially unsaturated 3-10 membered heterocyclyl), -C(=O)-C 1~6 Alkylene-(saturated or partially unsaturated 3-10 membered heterocyclyl), -C(=O)-(5-14 membered heteroaryl), -C(=O)-C 1~6 Alkylene-NH(C 1~6 alkyl), -C(=O)-C 1~6 Alkylene-N(C 1~6 Alkyl)2, N-methylpiperazine substituted acetyl, -S(=O)2R 1a , -P(=O)R 1a R 1b , [ka] is selected from the group consisting of However, R 1 and R 10 One of them is C 1~6 Alkyl and the other is H or C 3~10 When it is a cyclic hydrocarbyl, at least one of X and Y is a direct bond, ring C is not a 5-membered heteroaromatic ring, and R 1 and R 10 If one of is H and the other is: [ka] Ring C is not a 5-membered heteroaromatic ring, but R 1 and R 10 are both H, ring A contains at least one nitrogen atom and is not a 5- or 6-membered ring, and R 1 and R 10 If one of is H and the other is: [ka] Ring C is not a 5-membered heteroaromatic ring, but R 1 and R 10 is H and the other is H or acetyl, then ring D is absent; R 1a and R 1b are each independently H, halogen, amino, cyano, nitro, or C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~10 Cyclic hydrocarbyl, 3-10 membered heterocyclyl, C 6~10 Aryl, 5-14 membered heteroaryl, C 6~12 Aralkyl, -C(=O)R 5 , -OC(=O)R 5 , -C(=O)OR 5 , -OR 5 , -SR 5 , -S(=O)R 5 , -S(=O)2R 5 , -S(=O)2NR 5 R 6 , -NR 5 R 6 , -C(=O)NR 5 R 6 , -NR 5 -C(=O)R 6 , -NR 5 -C(=O)OR 6 , N.R. 5 -S(=O)2-R 6 , -NR 5 -C(=O)-NR 5 R 6 , -C 1~6 Alkylene-NR 5 R 6 , -C 1~6 Alkylene-OR 5 and -OC 1~6 Alkylene-NR 5 R 6 where R 1a and R 1b is n-propyl, the other is not H, or R 1a and R 1bform a 3- to 12-membered heterocyclic or heteroaromatic ring together with the atom to which they are attached; R 2 , R 3 , R 4 , R 7 , R 8 , R 9 and R 10 is represented by H, halogen, amino, cyano, nitro, C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~10 Cyclic hydrocarbyl, 3-10 membered heterocyclyl, C 6~10 Aryl, 5-14 membered heteroaryl, C 6~12 Aralkyl, -C(=O)R 5 ,OC(=O)R 5 , -C(=O)OR 5 , OR 5 , S.R. 5 , -S(=O)R 5 , -S(=O)2R 5 , -S(=O)2NR 5 R 6 , -NR 5 R 6 , -C(=O)NR 5 R 6 , -NR 5 -C(=O)R 6 , -NR 5 -C(=O)OR 6 , -NR 5 -S(=O)2-R 6 , -NR 5 -C(=O)-NR 5 R 6 , -C 1~6 Alkylene-NR 5 R 6 , -C 1~6 Alkylene -O(P=O)(OH)2 and -OC 1~6 Alkylene-NR 5 R 6 are each independently selected from the group consisting of: The above alkyl, alkylene, alkenyl, alkynyl, cyclic hydrocarbyl, hydrocarbon ring, heterocyclyl, heterocycle, aryl, aromatic ring, heteroaryl, heteroaromatic ring and aralkyl are each independently selected from the group consisting of halogen, hydroxyl, oxo, amino, cyano, nitro, C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~6 Cyclic hydrocarbyl, 3-10 membered heterocyclyl, C 6~10 Aryl, 5-14 membered heteroaryl, C 6~12 Aralkyl, =N-OR 5 , -C(=NH)NH2, -C(=O)R 5 , -OC(=O)R 5 , -C(=O)OR 5 , -OR 5 , -SR 5 , -S(=O)R 5 , -S(=O)2R 5 , -S(=O)2NR 5 R 6 , -NR 5 R 6 , -C(=O)NR 5 R 6 , -NR 5 -C(=O)R 6 , -NR 5 -C(=O)OR 6 , -NR 5 -S(=O)2-R 6 , -NR 5 -C(=O)-NR 5 R 6 , -C 1~6 Alkylene-NR 5 R 6 and -OC 1~6 Alkylene-NR 5 R 6 wherein the alkyl, cyclic hydrocarbyl, heterocyclyl, aryl, heteroaryl and aralkyl are each optionally substituted with one or more substituents independently selected from the group consisting of halogen, hydroxyl, oxo, amino, cyano, nitro, C 1~6 Alkyl, C 3~6 Cyclic hydrocarbyl, 3-10 membered heterocyclyl, C 6~10Aryl, 5-14 membered heteroaryl and C 6~12 and optionally further substituted with one or more substituents independently selected from the group consisting of aralkyl; R 5 and R 6 is H, alkyl, C 3~10 Cyclic hydrocarbyl, 3-10 membered heterocyclyl, C 6~10 Aryl, 5-14 membered heteroaryl and C 6~12 aralkyl, m, in each occurrence, is independently an integer of 0, 1, 2, or 3; n is an integer of 0, 1, or 2; i is an integer of 0, 1, or 2; g is an integer of 0, 1, 2, 3, or 4.

[0016] In one embodiment, the present disclosure provides compounds of formula II-IX: [ka] [ka] [ka] or a pharma- ceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotopically labeled compound thereof, 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 above.

[0017] In one embodiment, the present disclosure provides a compound of formula (X) or formula (XI): [ka] or a pharma- ceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotopically labeled compound thereof, wherein: R is H and C 1~6 is selected from the group consisting of alkyl, Ring D is a saturated or partially unsaturated 3- to 10-membered heterocycle, C 6~10 aryl, or a 5- to 10-membered heteroaromatic ring, preferably [ka] A phenyl ring, an N-methylpyrrole ring, a furan ring, or a thiophene ring. R 2 H and C 1~6 is selected from the group consisting of alkyl, R 3 , R 4 , R 7 , R 7’ and R 8 are H, halogen, -NH2, -OH, C at each occurrence. 1~6 Alkyl and -OR 5 are each independently selected from the group consisting of: R 9 and R 10 are H, halogen, and C in each occurrence. 1~6 Alkyl, C 2~6 Alkenyl, C 3~10 Cyclic hydrocarbyl, 3-10 membered heterocyclyl, C 6~10 Aryl, 5-14 membered heteroaryl, C 6~12 Aralkyl, -C(=O)R 5 , and -C 1~6 alkylene-O(P=O)(OH), The above alkyl, alkenyl, cyclic hydrocarbyl, heterocyclyl, aryl, heteroaryl, heteroaromatic and aralkyl are each independently selected from the group consisting of halogen, C 1~6 Alkyl and -OR 5 each optionally substituted with one or more substituents independently selected from the group consisting of: R 5 and R 6 In each occurrence, H, C 1~6 Alkyl, C 3~10 Cyclic hydrocarbyl, 3-10 membered heterocyclyl, C 6~10 Aryl, 5-14 membered heteroaryl and C 6~12 aralkyl, m, in each occurrence, is independently an integer of 0, 1, 2, or 3; n is an integer of 0, 1, or 2.

[0018] In a preferred embodiment, R 5 and R 6 is each occurrence independently selected from the group consisting of H, methyl and ethyl.

[0019] In a preferred embodiment, R 3 , R 4 , R 7 , R 7’ and R 8 is each occurrence independently selected from the group consisting of H, F, Cl, Br, -NH2, -OH, methyl, trifluoromethyl, -CH2-Ph, methoxy, ethoxy, and -CH2OCH3.

[0020] In a preferred embodiment, R 9 and R 10 stands for, at each occurrence, H, F, Cl, Br, methyl, ethyl, n-propyl, isopropyl, vinyl, cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl, monofluoromethyl, difluoromethyl, trifluoromethyl, acetyl, -OCH2CHF2, CH2OH, -CH2OCH3, -CH2CH2OCH3, -CH2-O(P=O)(OH)2, [ka] are each independently selected from the group consisting of:

[0021] In one embodiment, the present disclosure provides a compound of formula XII: [ka] or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotopically labeled compound thereof.

[0022] Compounds of formulas I-XII, in particular (6-(4-((4-(1H-pyrazol-4-yl)phenyl)amino)pyrimidin-2-yl)-1-methyl-1H-indol-2-yl)(3,3-difluoroazetidin-1-yl)methanone, can be prepared according to the methods disclosed in US2019 / 0276440, the entirety of which is incorporated herein by reference.

[0023] As used herein, the term "alkylene" refers to a saturated divalent hydrocarbyl, preferably having 1, 2, 3, 4, 5 or 6 carbon atoms, such as methylene (-CH-), ethylene (-CHCH-), propylene or butylene.

[0024] As used herein, the term "alkyl" is defined as a straight or branched chain saturated aliphatic hydrocarbon. In some embodiments, an alkyl has 1 to 12, e.g., 1 to 6, carbon atoms. For example, as used herein, "C 1~6The term "alkyl" refers to a straight or branched chain group having 1 to 6 carbon atoms (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, or n-hexyl, etc.), optionally substituted with one or more (e.g., 1 to 3) suitable substituents such as halogen (in which case the group is referred to as a "haloalkyl") (e.g., CH2F, CHF2, CF3, CCl3, C2F5, C2Cl5, CH2CF3, CH2Cl, or CH2CH2CF3, etc.). 1~4 The term "alkyl" refers to a straight 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).

[0025] As used herein, the term "alkenyl" refers to a straight or branched chain monovalent hydrocarbyl having a double bond and 2 to 6 carbon atoms ("C 2~6 Alkenyl refers to, 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 alkenylene group, the compounds may exist as pure E (entgegen) forms, pure Z (zusammen) forms, or any mixture thereof.

[0026] The term "alkynyl" as used herein 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.

[0027] As used herein, the term "cycloalkyl" refers to a saturated monocyclic or polycyclic (e.g., bicyclic) hydrocarbon ring (e.g., monocyclic, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, or cyclononyl, or bicyclic, including spiro, fused, or bridged ring systems (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.)), optionally substituted with one or more (e.g., 1 to 3) suitable substituents. Cycloalkyl has 3 to 15 carbon atoms. For example, "C 3~6 The term "cycloalkyl" refers to a saturated monocyclic or polycyclic (e.g., bicyclic) hydrocarbon ring having 3 to 6 ring-forming carbon atoms (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl), optionally substituted with one or more (e.g., 1 to 3) suitable substituents, such as methyl-substituted cyclopropyl.

[0028] As used herein, the terms "cyclic hydrocarbylene", "cyclic hydrocarbyl" and "hydrocarbon ring" refer to a monocyclic or polycyclic hydrocarbon ring that may be saturated (i.e., "cycloalkylene" and "cycloalkyl") or unsaturated (i.e., having one or more double and / or triple bonds within the ring), for example having 3 to 10 (suitably 3 to 8, more suitably 3 to 6) ring carbon atoms, including, but not limited to, cyclopropyl (cyclopropylene) (ring), cyclobutyl (cyclobutylene) (ring), cyclopentyl (cyclopentylene) (ring), cyclohexyl (cyclohexylene) (ring), cycloheptyl (cycloheptylene) (ring), cyclooctyl (cyclooctylene) (ring), cyclononyl (cyclononylene) (ring), cyclohexenyl (cyclohexenylene) (ring), and the like.

[0029] As used herein, the terms “heterocyclyl”, “heterocyclylene”, and “heterocycle” refer to, for example, a saturated (i.e., heterocycloalkyl) or partially unsaturated (having one or more double and / or triple bonds within the ring) cyclic group having 3 to 10 ring atoms (suitably having 3 to 8, more suitably 3 to 6), in which at least one ring atom is a heteroatom selected from the group consisting of N, O, and S, and the remaining ring atoms are C. For example, the “3- to 10-membered heterocyclyl(heterocyclylene)” of a “3- to 10-membered heterocycle” refers to a saturated or partially unsaturated heterocyclyl(heterocyclylene) 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 (oxiranylene), aziridinyl (aziridinylene), azetidinyl (azetidinylene), oxetanyl (oxetanylene), tetrahydrofuranyl (tetrahydrofuranylene), dioxolinyl (dioxolinylene), pyrrolidinyl (pyrrolidinylene), and pyrrolidonyl (pyrrolidonylene). , imidazolidinyl (imidazolidinylene), pyrazolidinyl (pyrazolidinylene), pyrrolinyl (pyrrolinylene), tetrahydropyranyl (tetrahydropyranylene), piperidinyl (piperidinylene), morpholinyl (morpholinylene), dithianyl (dithianylene), thiomorpholinyl (thiomorpholinylene), piperazinyl (piperazinylene) or trithianyl (trithianylene). The group also includes 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 heterocycles may be optionally substituted with one or more (e.g., 1, 2, 3, or 4) suitable substituents.

[0030] As used herein, the terms "aryl(arylene)" and "aromatic ring" refer to an all-carbon monocyclic or fused-ring polycyclic aromatic group having a conjugated pi-electron system. For example, as used herein, "C 6~10 Aryl(arylene)" and "C 6~10 The term "aromatic ring" refers to an aromatic group containing 6 to 10 carbon atoms, such as phenyl (phenylene) (benzene ring) or naphthyl (naphthylene) (naphthalene ring). The aryl (arylene) or aromatic ring may optionally contain one or more (e.g., 1 to 3) suitable substituents (e.g., halogen, -OH, -CN, -NO2, and C 1~6 It is substituted with alkyl.

[0031] As used herein, the terms "heteroaryl(arylene)" and "heteroaromatic ring" refer to monocyclic, bicyclic or tricyclic aromatic ring systems, especially having 5, 6, 8, 9, 10, 11, 12, 13 or 14 ring atoms, especially 1 or 2 or 3 or 4 or 5 or 6 or 9 or 10 carbon atoms, containing at least one heteroatom (such as O, N or S), which may be the same or different. Furthermore, in any case, they may be benzo-fused. In particular, the "heteroaryl(heteroarylene)" or "heteroaromatic ring" is selected from the group consisting of thienyl (thienylene), furyl (furylene), pyrrolyl (pyrrolylene), oxazolyl (oxazolylene), thiazolyl (thiazolylene), imidazolyl (imidazolylene), pyrazolyl (pyrazolylene), isoxazolyl (isoxazolylene), isothiazolyl (isothiazolylene), oxadiazolyl (oxadiazolylene), triazolyl (triazolylene), thiadiazolyl (thiadiazolylene), and the like, and benzo derivatives thereof, or pyridinyl (pyridinylene), pyridazinyl (pyridazinylene), pyrimidinyl (pyrimidinylene), pyrazinyl (pyrazinylene), triazinyl (triazinylene), and the like, and benzo derivatives thereof.

[0032] As used herein, the term "aralkyl" refers to an alkyl substituted with, preferably, an aryl or heteroaryl, where aryl, heteroaryl and alkyl are as defined herein. Typically, 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. Examples of aralkyl groups include, but are not limited to, benzyl, phenylethyl, phenylpropyl, and phenylbutyl.

[0033] The term “halo” or “halogen,” as used herein, is defined to include F, Cl, Br, or I.

[0034] As used herein, the term "nitrogen-containing heterocycle" refers to a saturated or unsaturated monocyclic or bicyclic group containing 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13 carbon atoms and at least one nitrogen atom in the ring, and may further optionally 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. The nitrogen-containing heterocycle is bonded to the remainder of the molecule through the nitrogen atom and any other ring atom of the nitrogen-containing heterocycle. The nitrogen-containing heterocycle is optionally benzo-fused, and is preferably bonded to the remainder of the molecule through the nitrogen atom of the nitrogen-containing heterocycle and any carbon atom of the fused benzene ring.

[0035] The term "substituted" means that one or more (e.g., 1, 2, 3, or 4) hydrogens on the specified atom are replaced with one selected from the specified group, provided that the replacement does not exceed the normal valence under the existing circumstances of the specified atom and that the substitution results in a stable compound. Combinations of substituents and / or variables are permissible only if such combinations result in stable compounds.

[0036] When a substituent is described as "optionally substituted," the substituent can be (1) unsubstituted or (2) substituted. When a carbon of a substituent is described as being optionally substituted with one or more of a set of substituents, one or more hydrogens on that carbon (if present) can be individually and / or simultaneously replaced with any independently selected substituents. When a nitrogen of a substituent is described as being optionally substituted with one or more of a set of substituents, one or more hydrogens on that nitrogen (if present) can each be replaced with any independently selected substituents.

[0037] When substituents are described as "independently selected" from a group, each substituent is selected independently of the other substituent(s). Thus, each substituent can be the same or different from the other substituent(s).

[0038] As used herein, the term "one or more" means one or more (eg, 2, 3, 4, 5 or 10), where appropriate.

[0039] As used herein, unless otherwise specified, the point of attachment of a substituent may be at any suitable position on that substituent.

[0040] When a bond to a substituent is shown as crossing a bond connecting two atoms in a ring, then such substituent may be bonded to any of the substitutable ring-forming atoms in that ring.

[0041] Compounds used in the methods provided herein include pharma- ceutically acceptable isotopically labeled compounds that are the same as those of Formulas I-XII except that one or more atoms have been replaced with an atom having the same atomic number, but the atomic mass or mass number is different from the atomic mass or mass number that predominates in nature. Examples of isotopes suitable for inclusion in the compounds include, but are not limited to, hydrogen isotopes, e.g. 2 H, 3 H, carbon isotopes, e.g. 11 C.13 C, and 14 Carbon isotopes such as C, chlorine isotopes, e.g. 36 Cl, fluorine isotopes, e.g. 18 F, iodine isotopes, e.g. 123 I and 125 I, nitrogen isotopes, e.g. 13 N and 15 N, oxygen isotopes, e.g. 15 O. 17 O and 18 O, phosphorus isotopes, e.g. 32 P, as well as sulfur isotopes, e.g. 35 Certain isotopically labeled compounds herein, for example those incorporating a radioactive isotope, are useful in drug and / or substrate tissue distribution studies (e.g., assays). 3 H), and carbon-14 (i.e. 14 C) are particularly useful for this purpose because of their ease of incorporation and rapidity of detection. 11 C. 18 F, 15 O and 13 Substitution with positron-emitting isotopes such as N can be useful in positron emission tomography (PET) studies to examine substrate receptor occupancy. Isotopically labeled compounds of the present disclosure can generally be prepared by processes similar to those described in the accompanying schemes and / or examples and preparations, by substituting the appropriate isotopically labeled reagent for the previously used non-isotopically labeled reagent. Pharmaceutically acceptable solvates according to the present disclosure include those in which the solvent of crystallization is isotopically substituted, for example, DO, acetone-d6, or DMSO-d6.

[0042] The term "stereoisomer" refers to an isomer 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 can exist as geometric isomers (cis / trans). Similarly, compounds provided herein can exist as mixtures of two or more structurally distinct forms (commonly referred to as tautomers) that are in rapid equilibrium. Representative examples of tautomers include keto-enol tautomers, phenol-keto tautomers, nitroso-oxime tautomers, imine-enamine tautomers, and the like. It will be understood that the use of all such isomers and mixtures thereof in any percentage (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, and 99%) are encompassed within the scope of the present disclosure.

[0043] The present disclosure includes the use of the crystalline forms or polymorphs of the compounds disclosed herein, either as a single polymorph or as a mixture of multiple polymorphs in any ratio.

[0044] It should also be understood that certain compounds provided herein can be used for treatment in free form or, if necessary, 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, which can directly or indirectly give the compounds of the present disclosure or their metabolites or residues after being administered to a patient in need of treatment.Therefore, compounds provided herein include various derivative forms of the above-mentioned compounds.

[0045] 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 pharma- ceutically 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, hybenate, hydrochloride / chloride, hydrobromide / bromide, hydroiodide / iodide, and the like. The salts include, isethionate, lactate, malate, maleate, malonate, mesylate, methylsulfate, naphthylate, 2-napsylate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate / hydrogenphosphate / dihydrogenphosphate, pyroglutamate, saccharate, stearate, succinate, tannate, tartrate, tosylate, trifluoroacetate and xinofoate. Suitable base addition salts are formed from bases which form pharma- ceutically acceptable salts. Specific non-limiting examples include aluminum, arginine, benzathine, calcium, choline, diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, potassium, sodium, tromethamine and zinc salts.

[0046] For a general discussion of suitable pharma- ceutical acceptable salts, see “Handbook of Pharmaceutical Salts: Properties, Selection, and Use” by Stahl and Wermuth (Wiley-VCH, 2002).

[0047] As used herein, the term "ester" refers to those derived from compounds of the various formulas provided herein, and includes physiologically hydrolyzable esters (which can be hydrolyzed under physiological conditions to release compounds of the present disclosure in the form of a free acid or alcohol).

[0048] The compounds for use in the methods of the present disclosure may exist as solvates (preferably hydrates), in which case the compounds contain a polar solvent, particularly water, methanol or ethanol, as a structural element of the compound's crystal lattice. The amount of polar solvent, particularly water, may be present in a stoichiometric or non-stoichiometric ratio.

[0049] As those skilled in the art will understand, not all nitrogen-containing heterocycles can form N-oxides, since nitrogen requires an available lone pair of electrons for oxidation to oxide. 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 preparing N-oxides of heterocycles and tertiary amines are well known to those skilled in the art, and include oxidizing heterocycles and tertiary amines with peracids 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 preparing N-oxides have been widely described and reviewed in the literature, see for example TLGilchrist, Comprehensive Organic Synthesis, vol. 7, pp 748-750, ARKatritzky and AJBoulton, Eds., Academic Press, and GWH Heeseman and ESG Werstiuk, Advances in Heterocyclic Chemistry, vol. 22, pp 390-392, ARKatritzky and AJBoulton, Eds., Academic Press.

[0050] The compounds described herein can be administered in the form of prodrugs, where certain derivatives of the compounds, which may themselves have little or no pharmacological activity, can be converted to compounds with the desired activity when administered into or onto the body, for example by hydrolytic cleavage. In general, such prodrugs are functional derivatives of compounds that are easily converted in vivo into compounds with 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). Prodrugs can be generated, for example, 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," for example as described in "Design of Prodrugs" by H. Bundgaard (Elsevier, 1985).

[0051] In some embodiments, the compounds of Formulae I-XII are selective inhibitors of Rho-associated coiled-coil kinase 2 (ROCK2) in human cells. The compounds of Formulae I-XII, e.g., as pharmaceutical compositions comprising the compounds, are used to treat (i.e., cure or reduce the severity of) viral infections, particularly coronavirus infections such as SARS-CoV-1, SARS-CoV-2, and MERS-CoV, and to treat or prevent sequelae 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 a MERS-CoV infection.

[0052] Disclosed herein are methods for determining kinase inhibition. 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 uses a fluorescently tagged peptide substrate. Phosphorylation of the tagged peptide by the kinase of interest promotes binding of the peptide to a trivalent metal-based nanoparticle via a specific high affinity interaction between the phosphate group and the trivalent metal. Approach to the nanoparticle results in an increase in fluorescence polarization. Inhibition of the kinase by a kinase inhibitor prevents phosphorylation of the substrate, thereby limiting binding of the fluorescently tagged substrate to the nanoparticle. Such an assay is compatible with a microwell assay format and allows for the determination of IC of multiple compounds. 50 can be measured simultaneously.

[0053] Pharmaceutical Compositions

[0054] In one aspect, the disclosure provides a pharma- ceutically acceptable composition for use in treating a viral disease, the composition comprising a therapeutically effective amount of one or more compounds of Formulas I-XII in combination with one or more pharma- ceutically acceptable carriers. As described in detail below, the pharmaceutical compositions of the present disclosure can be specifically formulated for administration in solid or liquid form, including those adapted for the following modes of administration: (1) oral administration, e.g., drenches (aqueous or non-aqueous solutions or suspensions), tablets (e.g., buccal, sublingual, and those intended for systemic absorption, boluses, powders, granules, pastes for application to the tongue, etc.; (2) parenteral administration, e.g., by subcutaneous, intramuscular, intravenous, or epidural injection, e.g., as a sterile solution or suspension, or as a sustained release formulation; (3) topical application, e.g., as a cream, ointment, or controlled release patch or spray applied to the skin; (4) vaginal or rectal administration, e.g., as a suppository, pessary, cream, or foam; (5) sublingual administration; (6) intraocular administration; (7) transdermal administration; or (8) nasal administration.

[0055] The phrase "pharmacologically acceptable" is used herein to refer to compounds, materials, compositions, and / or dosage forms that are suitable for use in contact with the tissues of human beings and animals, within the scope of sound medical judgment, without toxicity, irritation, allergic response, or other problem or complication commensurate with a reasonable benefit / risk ratio.

[0056] As used herein, the phrase "pharmaceutically acceptable carrier" refers to a pharma- ceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, manufacturing aid (e.g., lubricants, talc, magnesium, calcium, or zinc stearate, or stearic acid), solvent, or solvent encapsulating material involved in carrying or transporting a compound of interest from one organ or part of the body to another. Each carrier must be compatible with the other ingredients of the formulation and not deleterious to the patient. Some examples of materials that can function as pharma- ceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and cellulose derivatives, such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository wax; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; and (10) propylene glycol. (11) polyols, such as glycerin, sorbitol, mannitol, and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers, 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 buffered solutions; (21) polyesters, polycarbonates, and / or polyanhydrides; and (22) other non-toxic compatible substances used in pharmaceutical formulations.

[0057] The compounds of the present disclosure can be formulated using conventional carriers and excipients that can be selected according to ordinary practice. Tablets can include excipients, glidants, fillers, binders, and the like. Aqueous formulations can be prepared in a sterile form and generally be isotonic if intended for delivery by other than oral administration. Any formulation can optionally include excipients such as those described in the "Handbook of Pharmaceutical Excipients" (1986). Excipients include ascorbic acid and other antioxidants, chelating agents such as EDTA, carbohydrates such as dextran, hydroxyalkylcellulose, hydroxyalkylmethylcellulose, stearic acid, and the like.

[0058] While it is possible for the ROCK2 inhibitors disclosed herein (herein referred to as "active ingredients") to be administered alone, it may be preferable to present them as pharmaceutical formulations. The formulations of the present disclosure, for both veterinary and human use, comprise at least one active ingredient, as set forth above, together with one or more acceptable carriers for formulation and optionally other therapeutic ingredients, particularly the additional therapeutic ingredients discussed herein.

[0059] Formulations include those suitable for the routes of administration provided herein. Such formulations may conveniently be provided in unit dosage form and may be prepared by any method well known in the art of pharmacy. Methods and formulations are generally described in Remington's Pharmaceutical Sciences (Mack Publishing Co., Easton, Pa.). Such methods include the step of bringing into association the active ingredient with the carrier which constitutes one or more accessory ingredients. In general, formulations are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product.

[0060] Formulations of the present disclosure suitable for oral administration may be presented as discrete units such as capsules, cachets or tablets, each containing a predetermined amount of the active ingredient, as a powder or granules, as a solution or suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil emulsion. The active ingredient may also be administered as a bolus, electuary, or paste.

[0061] 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 the active ingredient in a free-flowing form, such as a powder or granules, optionally mixed with a binder, lubricant, inert diluent, preservative, surfactant or dispersant. Molded tablets can be prepared by molding in a suitable machine a mixture of powdered active ingredient moistened with an inert liquid diluent. Tablets can be optionally coated or scored, and are optionally formulated to provide slow or controlled release of the active ingredient from the tablet.

[0062] For infections of the eye or other external tissues, such as the mouth and skin, the formulations are preferably applied as topical solutions, ointments or creams containing the active ingredient(s). The active ingredient may be present in an amount of, for example, 0.075-20% w / w (including 0.1%-20% active ingredient(s) in the range of 0.1% w / w increments, such as, for example, 0.6% w / w, 0.7% w / w), preferably 0.2-15% w / w, most preferably 0.5-10% w / w. When formulated in an ointment, the active ingredient can be combined with either a paraffinic or water-miscible ointment base. Alternatively, the active ingredient can be formulated in a cream with an oil-in-water cream base.

[0063] Optionally, the aqueous phase of the cream base may contain at least 30% w / w of a polyhydric alcohol, i.e., an alcohol having two or more hydroxyl groups, such as propylene glycol, butane-1,3-diol, mannitol, sorbitol, glycerol, polyethylene glycol (including PEG 400), and mixtures thereof. Topical formulations may optionally contain a compound that enhances absorption or penetration of the active ingredient through the skin or other affected areas. Examples of such skin penetration enhancers include dimethylsulfoxide and related analogues.

[0064] The oily phase of the emulsion of the present invention can be composed of known ingredients in a known manner. This phase may only comprise an emulsifier (also known as an opacifier), but it is preferable to comprise a mixture of at least one emulsifier with fat or oil, or with both fat and oil. Preferably, a hydrophilic emulsifier is comprised together with a lipophilic emulsifier that acts as a stabilizer. It is also preferable to comprise both oil and fat. Taken together, the emulsifier(s) with or without stabilizer(s) constitute the so-called emulsifying wax, which together with oil and fat forms the oily dispersed phase of cream formulations, constitutes the so-called emulsifying ointment base.

[0065] 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. Additional emulsifiers and emulsion stabilizers suitable for use in the formulations of the present disclosure include Tween® 80.

[0066] The selection of suitable oils or fats for the formulation is based on whether the desired properties are achieved. The cream should preferably be a non-greasy, non-staining, washable product and should have an appropriate consistency to avoid leakage from tubes or other containers. Linear or branched monobasic or dibasic alkyl esters can be used, such as di-isoadipate, isocetyl stearate, propylene glycol diester of coconut fatty acid, isopropyl myristate, decyl oleate, isopropyl palmitate, butyl stearate, 2-ethylhexyl palmitate, or a blend of branched esters known as Crodamol CAP, the last three being the preferred esters. These can be used alone or in combination depending on the properties required. Alternatively, high melting point lipids such as white petrolatum and / or liquid paraffin, or other mineral oils, can also be used.

[0067] A pharmaceutical preparation according to the present disclosure includes a combination according to the present disclosure together with one or more pharma- ceutically acceptable carriers or excipients, and optionally other therapeutic agents. Pharmaceutical preparations containing active ingredients can 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. Compositions intended for oral use can be prepared according to any method known in the art to the manufacturer of pharmaceutical compositions, and such compositions may contain one or more agents, including sweeteners, flavoring agents, coloring agents, and preservatives, to provide a palatable preparation. Tablets containing the active ingredient in a mixture with non-toxic pharma- ceutically acceptable excipients suitable for the manufacture of tablets are acceptable. These excipients may be, for example, inert diluents such as calcium or sodium carbonate, lactose, calcium or sodium phosphate, granulating and disintegrating agents such as corn starch or alginic acid, binding agents such as starch, gelatin or acacia, and lubricants such as magnesium stearate, stearic acid or talc. The tablets may be uncoated or coated by known techniques, including microencapsulation to delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained action over a longer period. For example, a time delay material such as glyceryl monostearate or glyceryl distearate may be employed alone or with a wax.

[0068] Formulations for oral use may be presented as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent, such as starch, mannitol, calcium phosphate, or kaolin, or as soft gelatin capsules in which the active ingredient is mixed with water or an oil medium, such as peanut oil, liquid paraffin, or olive oil.

[0069] The aqueous suspensions of the present disclosure contain the active material mixed with excipients suitable for the manufacture of aqueous suspensions. Such excipients include suspending agents such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth, and gum acacia, dispersing or wetting agents such as naturally occurring phosphatides (e.g., lecithin), condensation products of alkylene oxides with fatty acids (e.g., polyoxyethylene stearates), condensation products of ethylene oxide with long chain aliphatic alcohols (e.g., heptadecaethyleneoxycetanol), condensation products of ethylene oxide with fatty acids and partial esters derived from hexitol anhydrides (e.g., polyethylene sorbitan monooleate). The aqueous suspensions may also contain one or more preservatives, for example, ethyl or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, such as sucrose or saccharin. Further non-limiting examples of suspending agents include cyclodextrins and Captisol (=sulfobutyl ether beta-cyclodextrin; SEB-β-CD).

[0070] Oily suspensions may be formulated by suspending the active ingredient in a vegetable oil, such as peanut oil, olive oil, sesame oil, or coconut oil, or in a mineral oil, such as liquid paraffin. Oral suspensions may also contain a thickening agent, such as beeswax, hard paraffin, or cetyl alcohol. Sweetening agents and flavoring agents, such as those described above, may be added to provide a palatable oral preparation. These compositions may be preserved by the addition of an antioxidant, such as ascorbic acid.

[0071] Dispersible powders and granules of the present disclosure suitable for preparing aqueous suspension by adding water provide active ingredient mixed with dispersing or wetting agent, suspending agent, and one or more preservatives.Suitable dispersing or wetting agent and suspending agent examples include those disclosed above.Additional excipients, such as sweeteners, flavoring agents, and coloring agents, may also be present.

[0072] The pharmaceutical composition of the present invention may be in the form of an oil-in-water emulsion. The oil phase may be a vegetable oil, such as olive oil or peanut oil, or a mineral oil, such as liquid paraffin, or a mixture thereof. Suitable emulsifiers may be naturally occurring gums, such as gum acacia and gum tragacanth, naturally occurring phosphatides, such as soybean lecithin, esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan monooleate, and the condensation products of these partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. The emulsion may also contain sweeteners and flavoring agents. Syrups and elixirs may be formulated with sweeteners, such as glycerol, sorbitol, or sucrose. Such formulations may also contain demulcents, preservatives, flavorings, or coloring agents.

[0073] The pharmaceutical compositions of the present disclosure may be in the form of a sterile injectable preparation, for example, a sterile injectable aqueous or oleaginous suspension. This suspension can be formulated according to known techniques using those suitable dispersing or wetting agents and suspending agents mentioned above. Sterile injectable preparations can also be prepared as sterile injectable solutions or suspensions in non-toxic parenterally acceptable diluents or solvents, such as solutions in 1,3-butanediol, or as lyophilized powders. Acceptable vehicles and solvents that can be used include water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile fixed oils are conventionally used as solvents or suspending media. For this purpose, any non-irritating fixed oil can be used, including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid can be used in the preparation of injectables as well. Acceptable vehicles and solvents that can be used include water, Ringer's solution, isotonic sodium chloride solution, and hypertonic sodium chloride solution.

[0074] The amount of active ingredient that can be combined with carrier materials to produce a single dosage form can vary depending on the host treated and the particular mode of administration. For example, a sustained release formulation intended for oral administration to humans can contain about 1-1000 mg of active ingredient, compounded with an appropriate and convenient amount of carrier material, which can vary from about 5 to about 95% (weight:weight) of the total composition. Pharmaceutical compositions can be prepared to provide easily measurable amounts for administration. For example, an aqueous solution intended for intravenous infusion can contain about 3-500 μg of active ingredient per milliliter of solution to result in infusion of a suitable volume at a rate of about 30 mL / hour.

[0075] Formulations suitable for topical administration to the eye also include eye drops in which the active ingredient is dissolved or suspended in a suitable carrier, especially an aqueous solvent for the active ingredient. The active ingredient may be present in such formulations in a concentration of 0.5 to 20% w / w, advantageously 0.5 to 10% w / w, especially about 1.5% w / w.

[0076] Formulations suitable for topical administration in the mouth include lozenges comprising the active ingredient in a flavored base, usually sucrose and acacia or tragacanth, pastilles comprising the active ingredient in an inert base such as gelatin and glycerin or sucrose and acacia, and mouthwashes comprising the active ingredient in a suitable liquid carrier.

[0077] Formulations for rectal administration may be presented as a suppository with a suitable base comprising, for example, cocoa butter or a salicylate.

[0078] Formulations suitable for pulmonary or nasal administration have, for example, particle sizes in the range of 0.1 to 500 microns (0.5, 1, 30, 35, etc.), which are administered by rapid inhalation through the nostrils to reach the alveolar sacs, or by inhalation through the mouth. Suitable formulations include aqueous or oily solutions of the active ingredient. Formulations suitable for aerosol or dry powder administration can be prepared according to conventional methods and can be delivered with other therapeutic agents, such as compounds.

[0079] Formulations suitable for vaginal administration may be presented as suppositories, pessaries, tampons, creams, gels, pastes, foams or spray formulations containing in addition to the active ingredient such carriers as are known in the art to be appropriate.

[0080] Formulations suitable for parenteral administration include aqueous and non-aqueous isotonic sterile injection solutions which may contain antioxidants, buffers, bacteriostats, and solutes which render the formulation isotonic with the blood of the intended recipient, and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents.

[0081] The formulations may be presented in unit-dose or multi-dose containers, for example, sealed ampoules and vials, and may be stored in a freeze-dried (lyophilized) condition for injection, requiring only the addition of a sterile liquid carrier, for example, water, immediately prior to use. Extemporaneous injection solutions and suspensions are prepared from sterile powders, granules and tablets of the kind previously described. Preferred unit dosage formulations are those containing a daily dose or unit daily sub-dose, as herein above recited, of the active ingredient, or an appropriate fraction thereof.

[0082] The disclosure further provides a veterinary composition comprising at least one active ingredient, as defined above, together with a veterinary carrier for the composition.

[0083] Veterinary carriers are materials useful for the purpose of administration of the compositions and may be otherwise inert or solid, liquid, or gaseous materials that are acceptable in the veterinary art and compatible with the active ingredient. These veterinary compositions may be administered orally, parenterally, or by any other desired route.

[0084] The compounds of the present disclosure are used to provide sustained release pharmaceutical formulations ("sustained release formulations") containing one or more compounds of the present disclosure as active ingredients, in which the release of the active ingredient is controlled and regulated to allow less frequent dosing or to improve the pharmacokinetic or toxicity profile of a particular active ingredient.

[0085] Combination with other activators

[0086] The ROCK2 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 signal transduction modulator, a β2-adrenergic receptor agonist bronchodilator, an anticholinergic agent, a mucolytic agent, a hypertonic saline solution, or a mixture thereof.

[0087] The compounds having Formulae I-XII can be administered in combination with one or more additional antiviral therapies and / or antiviral agents. The at least one other antiviral agent can be a nucleoside or nucleotide analog, or a pharma- ceutically 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, brivudine, acyclovir, ganciclovir, valacyclovir, cidofovir, valganciclovir, penciclovir, famciclovir, zidovudine, didanosine, zalcitabine, stavudine, abacavir, lamivudine, emtricitabine, tenofovir disoproxil fumarate, tenofovir alafenamide fumarate, adefovir, entecavir, telbivudine, sovosbuvir, and combinations or mixtures thereof.

[0088] Any compound of the present disclosure can be combined with one or more additional therapeutic agents as a single dosage form for simultaneous or sequential administration to patients.Combined therapy can be administered simultaneously or as a sequential regimen.When administered sequentially, the combination can be administered in two or more doses.

[0089] Co-administration of a compound of the present disclosure with one or more other active therapeutic agents generally refers to the simultaneous or sequential administration of a compound of the present disclosure and one or more other active therapeutic agents such that therapeutically effective amounts of both the compound of the present disclosure and the one or more other active therapeutic agents are present in the patient's body.

[0090] Concurrent administration includes administration of a unit dose of a compound of the present disclosure before or after administration of a unit dose of one or more additional therapeutic agents, for example, administration of a compound of the present disclosure within seconds, minutes, or hours of administration of one or more additional therapeutic agents. For example, a unit dose of a compound of the present disclosure can be administered within seconds or minutes after first administering a unit dose of one or more additional therapeutic agents. Alternatively, a unit dose of a compound of the present disclosure can be administered within seconds or minutes after first administering a unit dose of one or more additional therapeutic agents. In some cases, it may be desirable to administer a unit dose of a compound of the present disclosure a few hours (e.g., 1-12 hours) after first administering a unit dose of one or more additional therapeutic agents. In other cases, it may be desirable to administer a unit dose of a compound of the present disclosure a few hours (e.g., 1-12 hours) after first administering a unit dose of one or more additional therapeutic agents.

[0091] Combination therapy can provide "synergistic effects" and "synergistic effects", i.e., the effect obtained when each active ingredient is used together is greater than the sum of the effects resulting from using each compound separately. Synergistic effects can be obtained when each active ingredient is (1) co-formulated in a combined formulation and administered or delivered simultaneously, (2) delivered alternately or in parallel as separate formulations, or (3) by some other regimen. When delivered in alternation therapy, synergistic effects can be obtained when each compound is administered or delivered sequentially, for example, in separate tablets, pills, or capsules, or by different injection solutions in separate syringes. In general, in alternation therapy, an effective dose of each active ingredient is administered sequentially, i.e., sequentially, whereas in combination therapy, effective doses of two or more active ingredients are administered together. A synergistic antiviral effect refers to an antiviral effect that is greater than the expected purely additive effect of the individual compounds of the combination.

[0092] How to treat viral infections

[0093] As used herein, the terms "patient" and "subject" are used interchangeably.

[0094] In one aspect, the disclosure provides methods for treating viral infections, particularly coronavirus infections. In one aspect, the disclosure provides methods for treating and preventing sequelae of viral infections, including sequelae of coronavirus infections. In one aspect, the disclosure provides methods for preventing viral infections in subjects at risk for viral infections, including at risk for coronavirus infections.

[0095] In embodiments, the viral infection is 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, Arbovirus, Adenoviridae, Arenaviridae, Arterivirus, Astroviridae, Baculoviridae, Bimaviridae, Bunyaviridae, Caliciviridae, Calimoviridae, Circoviridae, Coronaviridae, Cystoviridae, EBV, Deltaviridae, Filoviridae, Flaviviridae, Iridoviridae, The viral infection is caused by a virus selected from the group consisting of mononegavirus, Myoviridae, Papillomavirus, Papovaviridae, Paramyxoviridae, prion, Parvoviridae, Phycodnaviridae, Poxviridae, Potyviridae, Reoviridae, Retroviridae, Rhabdoviridae, Tectiviridae, Togaviridae, smallpox, papilloma, influenza, Sendai virus (SeV), Sindbis virus (SINV), vaccinia virus, West Nile virus, hantavirus, viruses that cause the common cold, and any combination thereof. In an embodiment, the viral infection is caused by a coronavirus, such as SARS-CoV-1, SARS-CoV-2, and MERS-CoV. In an embodiment, the viral infection is caused by SARS-CoV-1. In an embodiment, the viral infection is caused by SARS-CoV-2. In embodiments, the viral infection is caused by one or more SARS-CoV-2 variants selected from the group consisting of alpha (B.1.1.7 and Q lineages), beta (B.1.351 and its descendant lineages), gamma (P.1 and its descendant lineages), epsilon (B.1.427 and B.1.429), eta (B.1.525), iota (B.1.526), ​​kappa (B.1.617.1), 1.617.3, mu (B.1.621, B.1.621.1), zeta (P.2), delta (B.1.617.2 and AY lineages), and omicron (B.1.1.529 and BA lineages).In an embodiment, the viral infection is caused by SARS-CoV-2 variant delta and / or SARS-CoV-2 variant omicron.

[0096] In an embodiment, the viral infection is caused by MERS-CoV.

[0097] In embodiments, the disclosure provides a method for treating or preventing one or more sequelae of COVID-19, comprising administering to a subject in need of treatment or prevention of one or more sequelae of COVID-19 a therapeutically effective amount of a compound of Formula I-XII. In embodiments, the disclosure provides a method for treating or preventing one or more sequelae of COVID-19, comprising administering to a subject in need of treatment or prevention of one or more sequelae of COVID-19 a therapeutically effective amount of a compound of Formula XII.

[0098] The sequelae of coronavirus infections, particularly SARS-CoV-1, SARS-CoV-2, and MERS-CoV, may result from one or more phenomena, such as organ damage from the acute infection phase, the development of a persistent hyperinflammatory state, continued viral activity associated with the host viral reservoir, or inappropriate antibody responses. COVID-19 sequelae include fatigue, dyspnea (difficulty breathing), cough, arthralgia (pain in the joints), and chest pain. Additional sequelae include cognitive impairment, depression, myalgia, headache, fever, and palpitations. COVID-19 sequelae may be cardiovascular (e.g., myocardial inflammation, ventricular dysfunction, stroke), respiratory (e.g., pulmonary function abnormalities, fibrosis), renal (e.g., acute kidney injury), dermatological (e.g., rash, alopecia), nervous (e.g., olfactory and gustatory dysfunction, sleep regulation disorders, cognitive changes, memory impairment), and / or psychiatric (e.g., depression, anxiety, mood changes) disorders.

[0099] Without wishing to be bound by theory, ROCK2 inhibitors, such as, for example, compounds of Formulas I-XII, are believed to block one or more of: (1) the pathways that viruses use to enter cells; (2) the cytoskeleton that viruses use to move and spread; (3) the pathways that viruses use to upregulate cellular energy metabolism; and (4) the pathways that viruses use to spread to other cells.

[0100] The ROCK2 inhibitors provided herein are believed to disrupt virus interaction with the host cell cytoskeleton, thereby inhibiting virus entry, replication, and / or spread. The cytoskeleton is a complex network of eukaryotic cells composed of three major types of cytoskeletal polymers, including actin filaments, microtubules, and intermediate filaments, that allows cells to perform multiple functions in a unified manner, such as connecting to the external environment, coordinating forces for movement and deformation, transporting vesicles through the cytoplasm, and spatially organizing contents. Most viruses hijack one or more aspects of the cytoskeletal network to facilitate their own infection. Viral interactions with host cell actin filaments, microtubules, and intermediate filaments play a key role in the life cycle of viruses, especially coronaviruses.

[0101] The ROCK2 inhibitors provided herein can interfere with the entry of viruses into target cells. After binding to target cells, viruses are believed to move to a suitable site for entry. It has been observed that when viruses reach the entry site, actin filaments contract and concentrate around the cell membrane. Pharmacological stabilization of the actin cortex can interfere with the entry of viruses into host cells. Viruses also utilize signaling pathways that regulate the cytoskeleton as part of the infection process.

[0102] Coronaviruses appear to utilize three cytoskeletal networks to complete the viral transport process: short-distance transport to and from the cell periphery is mediated by actin and its motor proteins, such as myosin, whereas long-distance transport is mediated by microtubules and the motor proteins dynein and kinesin. After entering the host cell, coronavirus-containing vesicles move along microtubules from the plasma membrane towards the replication site.

[0103] Cells infected with SARS-CoV-2 can fuse with neighboring cells to form actin-regulated syncytia. During the infection stage, SARS-CoV-2 migrates along filopodia on the host membrane to the site of cell entry and uses intermediate filament proteins to aid and utilize angiotensin-converting enzyme 2 (ACE2) receptors to enter target cells. In a mouse SARS model, SARS-CoV spike protein binds to ACE2 receptors and reduces ACE2 expression, resulting in severe lung injury. By binding to ACE2 receptors to gain cell entry, SARS-CoV-2 effectively inhibits ACE2 activity and the conversion of AT-II to angiotensin, leading to vasoconstriction, increased vascular permeability, adverse myocardial remodeling, and acute lung injury. Reduced expression of ACE2 and disruption of normal feedback loops may also lead to a lethal cytokine storm, as commonly seen in moderate to severe cases of COVID-19. Excessive inflammatory responses to SARS-CoV-2 are a major factor in severe illness and death in COVID-19 patients.

[0104] When ROCK2 is inhibited, ACE2 activity is restored, AT-II conversion continues, and proinflammatory cytokines (i.e., IL-1β, IL-6, and TNF-α), TGF-β1, and profibrotic markers (i.e., COL1A1, COL3A1, α-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 rearrangements, including migration, migration, survival, cell death, and formation of viral syncytia.

[0105] Furthermore, neurodevelopmental disorders and airway damage caused by coronaviruses are at least in part dependent on microtubules. Structural damage to the airway epithelium and abnormalities in cilia function are typical pathological symptoms of coronavirus infections. Cilia are composite structures composed of microtubules present on the cell surface. Coronaviruses that cause severe respiratory damage are thought to cause damage through the loss of cilia in the upper airways and lungs. Furthermore, microtubule disruption is thought to be associated with neurodegenerative diseases.

[0106] Based on their multifaceted mechanisms of action, ROCK2 inhibitors of formulae I-XII inhibit or prevent viral cell entry, for example by inhibiting syncytia, thus inhibiting viral spread. Secondly, ROCK2 inhibition also reduces the excessive immune response known as the "cytokine storm," as well as fibrotic changes in vascular, cardiac, and lung tissues that have led to long-term sequelae in some COVID-19 patients.

[0107] In some embodiments, the ROCK2 inhibitor (a) inhibits viral entry of a viral infection and / or (b) modulates viral entry of a viral infection.

[0108] In some embodiments, the ROCK2 inhibitor (a) inhibits viral spread of a viral infection and / or (b) modulates viral spread of a viral infection.

[0109] In some embodiments, ROCK2 inhibitor targets the target cell.Since ROCK2 inhibitor interacts with host cell target and not with viral target, the development of drug resistance to the effect of ROCK2 inhibitor in virus is expected to be delayed or substantially non-existent, when compared with other antiviral agents or compositions.In some embodiments, the use of ROCK2 inhibitor in the treatment of viral disease is characterized by the absence of drug resistance.

[0110] In some embodiments, a ROCK2 inhibitor inhibits and / or modulates the formation of stress fibers.

[0111] In some embodiments, the ROCK2 inhibitor inhibits actin filament dynamics and / or modulates actin filament dynamics.

[0112] In some embodiments, the ROCK2 inhibitor inhibits and / or modulates the mammalian target of rapamycin (mTOR) metabolic pathway.

[0113] In some embodiments, the ROCK2 inhibitor inhibits and / or modulates the AKT (serine / threonine protein kinase B) metabolic pathway.

[0114] In some embodiments, a ROCK2 inhibitor inhibits and / or modulates a membrane fusion pathway.

[0115] In some embodiments, the human subject is an adult patient. In embodiments, the human subject has or is at risk of having pulmonary fibrosis or scarring. In some embodiments, the human subject is a pediatric patient. In embodiments, the human subject has or is at risk of having Kawasaki disease or pulmonary fibrosis or scarring.

[0116] In embodiments, the ROCK2 inhibitor at least partially reverses and / or inhibits the level of fibrosis, at least partially inhibits excessive deposition of extracellular matrix in the lung, or improves blood supply to the lung. 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 lung pathology severity score associated with pulmonary fibrosis, (c) at least a 5% reduction in the expression of proinflammatory proteins, or (d) at least a 5% reduction in the expression of fibrogenic proteins.

[0117] In some embodiments, the human subject exhibits 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 antagonist to the human subject for at least two weeks. In some embodiments, the human subject does not exhibit a decrease in forced vital capacity after administration of the ROCK2 antagonist to the human subject for at least two weeks. In some embodiments, the human subject exhibits 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 a ROCK2 antagonist to the human subject for at least two weeks.

[0118] In some embodiments, the human subject shows a reduction in the incidence of coronary artery lesions in the first month of disease 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 administering the ROCK2 antagonist to the human subject for at least 2 weeks. In some embodiments, the ROCK2 antagonist at least partially reverses or reduces the vasculitis syndrome, or at least partially reverses or reduces the rash, redness of the eyes, lips, or tongue, swelling of the hands and feet, redness of the hands and feet, or swelling of the neck. In some embodiments, the ROCK2 antagonist rebalances the immune response in the human subject.

[0119] In one aspect, the disclosure provides a method of treating a patient suffering from a viral disease, comprising administering to a patient in need of such treatment a therapeutically effective amount of a compound of Formulas I-XII. As used herein, the phrase "therapeutically effective amount" refers to an amount of a compound, material, or composition comprising a compound of the disclosure effective to produce some desired therapeutic effect in at least a subpopulation of cells of a subject.

[0120] One or more compounds 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 appreciated that preferred routes may vary, for example, depending on the condition of the recipient.An advantage of the compounds of the present disclosure is that they are orally bioavailable and can be administered orally.

[0121] The effective dose of the compounds of the present disclosure depends at least on the nature of the condition being treated, the toxicity, whether the compound is used prophylactically or against an active viral infection, the delivery method, and the pharmaceutical formulation, and can be determined by the clinician using conventional dose escalation studies. In an embodiment, the dose of the compounds of the present disclosure ranges from about 0.1 to about 50 mg / kg of body weight per day. The daily dose for an adult can range from 1 mg to 1000 mg, for example, from about 5 mg to about 800 mg, or from about 50 mg to 500 mg, and can be in the form of a single or multiple doses per day. In an embodiment, the daily dose of a compound of formula I-XII for treating a viral infection or for treating or preventing one or more sequelae of an 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 may be administered as a single daily dose or divided into two, three or more daily doses. In an embodiment, the dose is about 200 mg total, administered once a day. In an embodiment, the dose is about 200 mg total, administered twice a day. In an embodiment, the dose is about 300 mg total administered once a day. In an embodiment, the dose is about 300 mg total administered twice a day. In an embodiment, the dose is about 400 mg total administered once a day. In an embodiment, the dose is about 400 mg total administered twice a day.

[0122] In embodiments, the compound of the present disclosure is first administered to the subject within about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 24, about 28, about 72, about 96, or about 120 hours after the subject is exposed to the virus. In embodiments, the compound of the present disclosure is first administered to the subject within about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 24, about 48, or about 72 hours after the subject develops symptoms in response to exposure to the virus. In embodiments, the compound of the present disclosure is first administered to the subject within about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 12, about 11, about 12, about 13, or about 14 days after the subject develops symptoms in response to exposure to the virus. In embodiments, the compounds of the present disclosure are administered prophylactically.

[0123] In embodiments, the compounds of the present disclosure are administered to a subject daily, every other day, every second day, every third day, once a week, twice a week, three times a week, once every two weeks, or once a month.

[0124] In embodiments, the disclosed compound is administered to the subject for about 1, about 2, about 3, about 4, about 5, about 6, or about 7 days. In embodiments, the disclosed compound is administered to the subject for about 1, about 2, about 3, about 4, about 5, about 6, or about 7, about 8, about 9, or about 10 weeks. In embodiments, the disclosed compound is administered to the subject for about 1 month, about 1.5 months, about 2 months, about 2.5 months, about 3 months, about 4 months, about 5 months, about 6 months, or more.

[0125] In embodiments, the compound of the present disclosure is administered to the subject about 1, about 2, about 3, about 4, about 5, about 6, or about 7 days after the subject is first exposed to the virus. In embodiments, the compound of the present disclosure is administered to the subject about 1, about 2, about 3, about 4, about 5, about 6, or about 7, about 8, about 9, or about 10 weeks after the subject is first exposed to the virus. In embodiments, the compound of the present disclosure is administered to the subject about 1 month, about 1.5 months, about 2 months, about 2.5 months, about 3 months, about 4 months, about 5 months, about 6 months, or more after the subject is first exposed to the virus.

[0126] In embodiments, the compound of the present disclosure is administered to the subject about 1, about 2, about 3, about 4, about 5, about 6, or about 7 days after the subject develops symptoms in response to exposure to the virus. In embodiments, the compound of the present disclosure is administered to the subject about 1, about 2, about 3, about 4, about 5, about 6, or about 7, about 8, about 9, or about 10 weeks after the subject develops symptoms in response to exposure to the virus. In embodiments, the compound of the present disclosure is administered to the subject about 1 month, about 1.5 months, about 2 months, about 2.5 months, about 3 months, about 4 months, about 5 months, about 6 months, or more after the subject develops symptoms in response to exposure to the virus.

[0127] In embodiments, the compound of the present disclosure is administered to the subject about 1, about 2, about 3, about 4, about 5, about 6, or about 7 days after the subject has recovered from symptoms in response to exposure to the virus. In embodiments, the compound of the present disclosure is administered to the subject about 1, about 2, about 3, about 4, about 5, about 6, or about 7, about 8, about 9, or about 10 weeks after the subject has recovered from symptoms in response to exposure to the virus. In embodiments, the compound of the present disclosure is administered to the subject about 1 month, about 1.5 months, about 2 months, about 2.5 months, about 3 months, about 4 months, about 5 months, about 6 months, or more after the subject has recovered from symptoms in response to exposure to the virus.

[0128] Treatment of Kawasaki disease / viral infection-related symptoms

[0129] Some pediatric patients with COVID-19 present with pediatric inflammatory multisystem 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 vasculitis syndromes or inflammatory diseases that cause inflammation of blood vessels and in some cases swelling throughout the body, including Kawasaki disease-like diseases such as PMIS or PIMS-TS. Symptoms of Kawasaki disease may include a high fever (>101°F) lasting at least four days, as well as a rash, redness of the eyes, lips / tongue, swelling and redness of the hands and feet, and swelling of the neck. As used herein, the term "Kawasaki disease shock syndrome" or KDSS refers to Kawasaki disease patients who exhibit a greater than 20% decrease in systolic blood pressure compared to age-matched healthy subjects or who exhibit peripheral blood circulation disorders. KDSS may be due to high levels of circulating inflammatory factors.

[0130] Pediatric patients with Kawasaki disease associated with COVID-19, PMIS, and PIMS-TS may have increased inflammatory markers, such as neutrophil-predominant leukocytosis, C-reactive protein (CRP), procalcitonin (PCT), and IL-6. The development of COVID-19-associated Kawasaki disease or PMIS may be caused by immunological responses following viral infection, such as antibody or immune complex-mediated responses.

[0131] Kawasaki disease, a type of systemic vasculitis, is thought to involve small to medium-sized blood vessels. The most serious complications or sequelae are myocardial infarction, coronary artery fistula, coronary artery dilatation, and the formation of coronary artery lesions (CALs), such as coronary artery aneurysms, which may subsequently lead to long-term sequelae such as stenosis or occlusion, and myocardial infarction. Therefore, a reduction in the incidence of coronary artery aneurysms in patients with Kawasaki disease is thought to be an indicator of the effectiveness of treatment.

[0132] ROCK2 inhibitors of Formulas I-XII can reduce peripheral blood levels of proinflammatory cytokines such as IL-17 and IL-23. Additionally, these ROCK2 inhibitors are believed to rebalance the immune response in pediatric patients with Kawasaki Disease, PMIS, or PIMS-TS following infection with COVID-19 or other coronaviruses.

[0133] Method for treating fibrosis caused by viral infection - Patents.com

[0134] COVID-19 patients may develop scarring in the lungs during the acute phase of the disease or after recovery from the disease. The lung scarring may be fibrotic or fibrotic scarring. The underlying causes and clinical course of lung scarring in COVID-19 patients may be the same as or different from patients with interstitial lung disease and pulmonary fibrosis such as idiopathic pulmonary fibrosis (IPF) or rheumatoid arthritis.

[0135] Fibrosis is the hyperproliferation, hardening, and / or scarring of various tissues and is believed to result from excessive deposition of extracellular matrix components such as collagen. Fibrosis is believed to be the result of chronic inflammatory responses triggered by a variety of stimuli including, for example, persistent infection, autoimmune responses, allergic responses, chemical insults, radiation, and tissue damage.

[0136] COVID-19 infection can cause a variety of respiratory diseases, ranging from atypical pneumonia to acute respiratory distress syndrome (ARDS). Many patients infected with the virus may share characteristics with patients suffering from idiopathic pulmonary fibrosis (IPF). In IPF patients, lung function inevitably declines, which can lead to respiratory failure and death. A possible cause of lung damage is cytokine release syndrome triggered by viral antigens.

[0137] The ROCK2 signaling pathway controls cell motility and shape. In addition, ROCK2 appears to regulate cytokine secretion in T cells, for example promoting proinflammatory cytokines such as IL-17 and IL-21, whereas the secretion of anti-inflammatory cytokines IL-2 and IL-10 is negatively regulated by ROCK2 under Th17 skewing activation. ROCK2 also contributes to the control of IFN-γ secretion in T cells from rheumatoid arthritis patients in disease but not in steady state. Therefore, ROCK2 signaling is a key pathway in the regulation of T cell-mediated immune responses, highlighting the therapeutic potential of targeted inhibition of ROCK2 in autoimmunity. Thus, ROCK2 inhibitors can reduce the secretion of IL-21 and IL-17 in human T cells via a STAT3-dependent mechanism.

[0138] ROCK2 inhibitors of formulas I-XII can reduce peripheral blood levels of IL-17 and IL-23, two pro-inflammatory cytokines. In addition, these ROCK2 inhibitors can simultaneously increase the immunosuppressive cytokine IL-10 and increase the percentage of Foxp3+CD4 T cells in the blood, which is believed to reduce the immune inflammatory response. Therefore, ROCK2 inhibitors with formulas I-XII can rebalance the immune response in patients suffering from fibrosis after infection with SARS-CoV-2 or other coronaviruses.

[0139] Many pulmonary function parameters can be used to determine the effective amount of a ROCK2 inhibitor, i.e., the amount that reduces, stabilizes, or reverses the rate of pathological decline in one or more pulmonary function parameters in patients suffering from a viral infection, such as a coronavirus infection, or to monitor the patient's response to ROCK2 inhibitor therapy. These pulmonary function parameters include vital capacity (VC), forced expiratory volume (FEV), forced vital capacity (FVC), and FVC%.

[0140] As used herein, the term "vital capacity" (VC) refers to the total volume of air that can enter and leave the lungs. VC is equal to the combined inspiratory reserve, tidal volume, and expiratory reserve volume.

[0141] As used herein, the term "forced expiratory volume" (FEV) refers to measuring the amount of air a subject can exhale during forced breathing. The amount of exhaled air can be measured during the first second (FEV1), second (FEV2), and / or third (FEV3) of forced breathing. For example, the FEV1 / FVC ratio refers to the ratio of forced expiratory volume in 1 second to forced vital capacity.

[0142] As used herein, the term "forced vital capacity" (FVC) refers to maximal expiratory effort vital capacity, i.e., the total volume of air exhaled by a subject during an FEV test.

[0143] As used herein, the term "FVC%" refers to the rate of change of a subject's FVC over a period of time.Predicted FVC% is the measured FVC of a subject expressed as a percentage of the predicted FVC of the subject.As used herein, all FVC% predicted values ​​are absolute values, not relative values.

[0144] Many of these lung function parameters can be readily obtained using a spirometer. Residual volume can be obtained by indirect methods such as x-ray planimetry, body plethysmography, closed-circuit dilution (including helium dilution), and nitrogen sparging.

[0145] Lung volumes and related pulmonary function parameters naturally decline with age. A number of studies have been conducted in normal populations to measure the rate of decline in lung volumes 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 can expect to experience a decline in FVC of 0.03 liters at age 66.

[0146] In contrast to the natural decline with age, subjects with lung diseases such as pulmonary fibrosis and fibrosis due to COVID-19 infection experience an abnormally steep rate of decline in lung volume or one or more lung function parameters, a so-called "pathological rate of decline." As used herein, a "pathological rate of decline" is a rate of decline in lung volume or one or more lung function parameters that is at least 5% greater than the normal decline with age. 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% greater than the expected rate of decline in a normal person similarly matched for race or ethnicity, sex, age, height, and weight. The rate of decline can be expressed as a change from baseline per week, 2 weeks, 4 weeks, 8 weeks, 12 weeks, 24 weeks, 36 weeks, 48 ​​weeks, or 12 months. In certain embodiments, the pathological rate of decline in lung volume 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 12 months. In other embodiments, the pathological rate of decline is a change from baseline percent predicted forced vital capacity (FVC%) of at least about -2%, -3%, -4%, -5%, -6%, -7%, -8%, or -10% per 12 months.

[0147] Administration of a ROCK2 inhibitor of formula I-XII can result in an increase in FVC in subjects with fibrosis after treatment compared to before treatment. Treatment with an effective amount of a 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 FVC before treatment. In some embodiments, treatment with a ROCK2 inhibitor lasts 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 extends for 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.

[0148] Methods for Treating Arenavirus Infections

[0149] In embodiments, the present application provides a method of treating an Arenaviridae virus infection in a human, comprising administering to a patient in need of treatment for an Arenaviridae virus infection a therapeutically effective amount of a ROCK2 inhibitor disclosed herein, or a pharma- ceutically acceptable salt, solvate, and / or ester thereof, In embodiments, at least one additional active therapeutic agent is administered to the patient.

[0150] Also provided is the use of a ROCK2 inhibitor, or a pharma- ceutically acceptable salt, solvate, and / or ester thereof, in the preparation of a medicament for use in the treatment of an Arenaviridae infection in a human.

[0151] Arenaviruses are single-stranded negative-sense RNA viruses that typically infect primates. Arenaviruses can grow in virtually all cell types. Based on studies in non-human primates infected with Lassa virus, the first cells to be infected appear to be dendritic cells in lymphoid tissues. Infection progresses to infection of Kupffer cells in the liver, parenchymal cells in the liver and adrenal gland, endothelial cells in various tissues including neural tissue, and ultimately infection of the epithelium. Evidence of human liver infection causing hepatitis has also been described in the literature (Hensley, L., 2011, Virology Journal; Yun, NE, 2012 Viruses).

[0152] Arenaviruses include Alpawayovirus (ALLV), Amaparivirus (AMAV), Bear Canyon virus (BCNV), Catalinavirus, Chaparevirus, Cupicivirus (CPXV), Dandenong virus, Flexar virus (FLEV), Guanaritovirus (GTOV), Yippee virus (IPPYV), Junin virus (JUNV), Codcovirus, Lassa virus (LASV: 6 strains, Josiah, NL, z148, Macenta, AV, and CSF), Latinovirus (LATV), and lymphocytic pulse virus (LVV). Thirty genera have been identified: cholangiocarcinoma virus (LCMV), Rujo virus, Machupo virus (MACV), Mobara virus (MOBV), Morogoro virus, Mopeia virus (MOPV), Oliveros virus (OLVV), Parana virus (PARV), Pichinde virus (PICV), Pinar virus, Piritar virus (PIRV), Sabia virus (SABV), Skinner tank virus, Tacaribe virus (TCRV), Tamiami virus (TAMV), or Whitewater Arroyo virus (WWAV).

[0153] In some embodiments, the compound of the present disclosure is administered in combination with an additional therapeutic agent. In the treatment of Arenaviridae virus infection, the additional therapeutic agent is preferably active against Arenaviridae virus infection, particularly Lassa virus infection and Junin virus infection. Non-limiting examples of the additional therapeutic agent include ribavirin, favipiravir (also known as T-705 or Avigan), T-705 monophosphate, T-705 diphosphate, T-705 triphosphate, ST-193, and mixtures thereof. The compounds and compositions of the present disclosure are also intended for use with common treatments given to patients with Arenaviridae virus infections, including parenteral fluids (including dextrose saline and lactated Ringer's solution) and nutrition, antibiotics (including metronidazole and cephalosporin antibiotics such as ceftriaxone and cefuroxime) and / or antifungal prophylaxis, antipyretic and analgesic drugs, antiemetics (such as metoclopramide) and / or antidiarrheal drugs, vitamin and mineral supplements (including vitamin K and zinc sulfate), anti-inflammatory drugs (such as ibuprofen), analgesics, and drugs to treat other common illnesses in the patient population, such as antimalarials (including artemether and artesunate-lumefantrine combination therapy), typhoid fever (including quinolone antibiotics such as ciprofloxacin, macrolide antibiotics such as azithromycin, cephalosporin antibiotics such as ceftriaxone, or aminopenicillins such as ampicillin), or shigellosis.

[0154] The compounds of the present disclosure are useful for the treatment or prevention of Arenaviridae infections in animals or humans.

[0155] However, when screening for compounds capable of inhibiting Arenaviridae, it is important to keep in mind that the results of enzyme assays may not correlate with those of cell culture assays, and therefore cell-based assays should be the primary screening tool.

[0156] In another aspect, the application provides a method of treating an Arenaviridae virus infection in a human, comprising administering to a patient a therapeutically effective amount of a ROCK2 inhibitor disclosed herein, or a pharma- ceutically acceptable salt, solvate, and / or ester thereof. In some embodiments, the Arenaviridae infection is caused by a virus of the Arenaviridae family. In some embodiments, the Arenaviridae infection is caused by Junin virus. In some embodiments, the Arenaviridae infection is caused by Lassa virus strain Josiah, NL, z148, Macenta, AV, or CSF.

[0157] The compounds of the present disclosure can be used to treat humans already suffering from Arenaviridae infection or can be administered prophylactically to reduce or prevent the possibility of Arenaviridae infection. A post-fever physical examination of patients infected with Arenaviruses often reveals purulent pharyngitis, bilateral conjunctival hemorrhages, facial edema, and generalized abdominal tenderness. Gross pathological changes include pleural effusion, pulmonary edema, ascites, and hemorrhagic symptoms of the gastrointestinal mucosa. Mortality in hospitalized patients varies from 5-10%.

[0158] kit

[0159] In one aspect, the disclosure provides a kit or composition comprising a compound of Formula I-XII, or a pharma- ceutically acceptable salt, pharma- ceutically acceptable ester, stereoisomer, mixture of stereoisomers, or tautomers thereof. In another embodiment, separate kits are provided comprising a compound of Formula I-XII, or a pharma- ceutically acceptable salt, pharma- ceutically acceptable ester, stereoisomer, mixture of stereoisomers, or tautomers thereof. In one aspect, the kit comprises a compound of Formula I-XII, or a pharma- ceutically acceptable salt thereof. Each of the separate kits described herein can include a label and / or instructions for use of the compound in treating 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 some embodiments, the disease or condition is a human Arenaviridae virus infection, including a Lassa virus infection or a Junin virus infection. In other embodiments, each separate kit can include instructions for use of an additional agent in combination with a compound of Formula I-XII in treating a disease or condition in a subject (e.g., a human) in need thereof. In some embodiments, the kit comprises individual dosage units of the compounds described herein.Examples of individual dosage units include pills, tablets, capsules, pre-filled syringes or syringe cartridges, IV bags, etc., each of which comprises a therapeutically effective amount of the compound, or its pharma- ceutically acceptable salts, racemates, enantiomers, diastereomers, tautomers, polymorphs, pseudopolymorphs, amorphous forms, hydrates or solvates.In some embodiments, the kit may comprise a single dosage unit, and in other cases, there are multiple dosage units, such as the number of dosage units required for a particular regimen or period.

[0160] Also provided is an article of manufacture comprising a compound of Formula I-XII, or a pharma- ceutically acceptable salt, pharma- ceutically acceptable ester, stereoisomer, mixture of stereoisomers, or tautomers thereof, and a container. In one aspect, the article of manufacture comprises a compound of Formula I-XII, or a pharma- ceutically acceptable salt thereof, and a container. In some embodiments, the article of manufacture container is a vial, jar, ampoule, pre-filled syringe, blister package, tin, can, bottle, box, or intravenous bag. EXAMPLES

[0161] Example 1: In vitro SARS-CoV-2 prophylactic model A 50 mM stock solution of (6-(4-((4-(1H-pyrazol-4-yl)phenyl)amino)pyrimidin-2-yl)-1-methyl-1H-indol-2-yl)(3,3-difluoroazetidin-1-yl)methanone (Formula XII) in DMSO was diluted with PBS to concentrations of 100 μM, 10 μM, 1 μM, 0.3 μM, 0.1 μM, and 0.03 μM of Formula XII. Approximately 30,000 immortalized human airway epithelial cells (Calu-3) were grown in a 96-well format to approximately 100,000 cells per well. Cells were treated with a compound of Formula XII or a control by adding 10 μl of Formula XII solution (0.03-100 μM) or control solution (DMSO in PBS) in triplicate and incubating at 37° C. for 2 hours. Cells were then incubated for 2 hours at 37° C. ... 2 Cells were challenged with PFU of SARS-CoV-2 (strain 2019-nCoV / USA-WA1 / 2020) and incubated for 6 h at 37 °C, followed by washing. Positive controls consisted of alisporivir (5 μM). Further controls (3 wells) were treated with the highest concentration of Formula XII, but were not challenged with virus. Cells were maintained until the 48-h endpoint.

[0162] Viral infection was analyzed by qPCR (qRT-PCR). RNA from cells was extracted using TRIzol reagent (Thermo Fisher) according to the manufacturer's instructions. Levels of viral or host RNA in the supernatants were measured using TaqPath™ 1-Step RT-qPCR Master Mix (Thermo Fisher) on a CFX Connect Real-Time System (Bio-Rad) instrument with the following primers: forward: 5'-GACCCCAAAATCAGCGAAAT-3', reverse: 5'-TCTGGTTACTGCCAGTTGAATCTG-3'.

[0163] Cytotoxicity was measured by LDH (lactate dehydrogenase) assay (Clontech) according to the manufacturer's instructions. The positive control consisted of saponin (0.5%).

[0164] Each data set was analyzed and plotted with GraphPad Prism (version 9.0.2). Simulations of dose-response curve analysis using the median effect principle and mass action law were performed using CompuSyn (https: / / www.combosyn.com / ). The results of the in vitro studies are shown in Figures 1-5.

[0165] Example 2. Formula XII reduces the spread of SARS-CoV-2 in the lungs in vivo Human ACE2 transgenic mice (n=5 per group) obtained from Jackson Laboratories were challenged at t=0 with an initial viral load of 100 PFU, 1,000, or 10,000 pfu of the 2019-nCoV / USA-WA1 / 2020 strain of SARS-CoV-2. The strain was isolated from an oropharyngeal swab of a patient with respiratory disease who developed clinical disease (COVID-19) in Washington State, USA in January 2020 (BEI Resources, NR-52281). A viral load of 10,000 pfu is considered to be significantly higher than the corresponding viral load occurring naturally in human subjects. Infected mice were administered daily by oral gavage with (1) 0.5% sodium carboxymethylcellulose (CMC-Na) (negative control), (2) 300 mg / kg Formula XII in 0.5% CMC-Na (test group), or (3) Remdesivir (RDV). RDV, an antiviral reagent that acts as a nucleoside analogue and inhibits the RNA-dependent RNA polymerase (RdRp) of coronaviruses, including SARS-CoV-2, was used as a positive control. RDV was solubilized at 2.5 mg / mL in a vehicle containing 12% sulfobutylether-β-cyclodextrin sodium salt (with HCl / NaOH) in water, pH 5.0 (25 mg / kg subcutaneously) and continued daily until the end of the study. Animals were monitored daily. Two animals were sacrificed at the indicated times (Figure 6). Viral load in the lungs was measured by qPCR detecting viral RNA in lung lysates.

[0166] As shown in FIG. 6, treatment with Formula XII resulted in a significant reduction in viral load compared to both the negative and positive controls.

[0167] Example 3. Treatment of COVID-19 with ROCK2 inhibitors The study will be conducted in men and women suspected of having or with SARS-CoV-2 infection. The study will use (6-(4-((4-(1H-pyrazol-4-yl)phenyl)amino)pyrimidin-2-yl)-1-methyl-1H-indol-2-yl)(3,3-difluoroazetidin-1-yl)methanone (Formula XII) administered orally at 100-400 mg daily for a treatment period of 2-3 weeks. Different amounts of Formula XII will be administered at specified intervals.

[0168] Subjects are observed for about 2 weeks to about 6 months. Coronavirus testing is performed daily, every 2 days, every 3 days, or other intervals during the treatment period by collecting samples from the patient using a nasopharyngeal swab, oropharyngeal swab, mid-nasal turbinate swab, or anterior nares swab. The swab is then immediately placed in a sterile transport tube containing about 2-3 mL of either viral transport medium (VTM), Amies transport medium, or sterile saline, unless a coronavirus point-of-care test is used. The sample is tested using a CDC-approved testing method. Different doses of Formula XII administered at 100 mg / day, 200 mg / day, 300 mg / day, or 400 mg / day are also tested in patients following the same protocol, and the dose-response relationship is recorded for the degree of viral inhibition.

[0169] Example 4. Treating COVID-19 with a ROCK2 inhibitor and remdesivir The study will be conducted in men and women suspected of having or with SARS-CoV-2 infection. The study will use Formula XII, administered orally at 100-400 mg daily for a treatment period of 2-3 weeks. In parallel, remdesivir will be administered to adult patients by intravenous infusion over 30-120 minutes in a total volume of 250 mL of 0.9% saline according to the following schedule for adult and pediatric patients weighing 40 kg or more: a loading dose of 200 mg on day 1, followed by approximately 100 mg once daily on days 2-10.

[0170] Subjects are observed for about 2 weeks to about 6 months. Coronavirus testing is performed daily, every 2 days, every 3 days, or other intervals during the treatment period by collecting samples from the patient using a nasopharyngeal swab, oropharyngeal swab, mid-nasal turbinate swab, or anterior nares swab. The swab is then immediately placed in a sterile transport tube containing about 2-3 mL of either viral transport medium (VTM), Amies transport medium, or sterile saline, unless a coronavirus point-of-care test is used. The sample is tested using a CDC-approved testing method. Different doses of Formula XII administered at 100 mg / day, 200 mg / day, 300 mg / day, and 400 mg / day are also tested in patients following the same protocol, and the dose-response relationship is recorded for the degree of viral inhibition.

[0171] Example 5. Treatment of fibrosis using ROCK2 inhibitors The study is conducted in 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 2. In addition to examining the extent 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, 3, 4, or 5 days, weekly, biweekly, or monthly, to monitor the progression of fibrosis. If clinically approved, X-rays and / or CT scans can be performed. Furthermore, the period of administering Formula XII can be extended beyond 2 weeks at different dose levels in this study.

[0172] To assess each subject's pulmonary function following treatment with Formula XII, 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 days 1, 3, 7, 14, monthly, or bimonthly of treatment.

[0173] Example 6. Treatment of fibrosis using ROCK2 inhibitors and remdesivir The study is conducted on 50 men and women suspected of having SARS-CoV-2 infection or having SARS-CoV-2 infection and suspected of having or having fibrosis in the same manner as described in Example 2. In addition to examining the extent 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, 3, 4, or 5 days, weekly, biweekly, or monthly, to monitor the progression of fibrosis. If clinically approved, X-rays and / or CT scans can be performed. Furthermore, the period of administering Formula XII can be extended beyond 2 weeks at different dose levels in this study.

[0174] To assess each subject's pulmonary function following treatment with Formula XII, 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 days 1, 3, 7, 14, monthly, or bimonthly of treatment.

[0175] Example 7. Treatment of Kawasaki disease-related disorders using ROCK2 inhibitors The study will be conducted on pediatric patients suspected of having SARS-CoV-2 infection or suspected of having Kawasaki disease-like disease or with pediatric multisystem inflammatory syndrome (PMIS) or pediatric inflammatory multisystem syndrome temporally associated with SARS-CoV-2 (PIMS-TS). The study will use Formula XII, which will be administered orally at 100 mg / day, 200 mg / day, 300 mg / day, and 400 mg / day. Different amounts of Formula XII will be administered at specified intervals.

[0176] Subjects are observed for approximately 2 weeks to approximately 6 months. Coronavirus testing is performed daily, every 2 days, every 3 days, or other intervals during the treatment period by collecting specimens from patients using a nasopharyngeal swab, oropharyngeal swab, mid-nasal turbinate swab, or anterior nares swab. The swab is then immediately placed into a sterile transport tube containing approximately 2-3 mL of either viral transport medium (VTM), Amies transport medium, or sterile saline, unless a coronavirus point-of-care test is used. Samples are tested using a CDC-approved testing method. Different doses of Formula XII 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 following the same protocol and the dose-response relationship for the degree of viral inhibition is recorded.

[0177] In addition to testing the extent of viral infection, vasculitis tests, such as blood tests (for C-reactive protein, complete blood count, amount of antineutrophil cytoplasmic antibodies, or other biomarkers), urinalysis (red blood cell or urinary protein content), and 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, 3, 4, or 5 days, weekly, biweekly, or monthly, to monitor the progression of Kawasaki-like disease, PMIS, or PIMS-TS. Furthermore, the period of administration of Formula XII can be extended beyond 2 weeks at different dose levels in this study.

[0178] To assess disease progression, patients undergo a 2-dimensional echocardiogram one month after the disease to evaluate coronary artery lesions (CAL). Measurements for each patient include the diameters of the left main coronary artery (LMCA), left anterior descending artery (LAD), left circumflex artery (LCX), and proximal and mid-right coronary artery (RCA). A Z-score is calculated for each coronary artery. See Journal of the American Society of Echocardiography, 2011, 24(1). CAL can be defined as a z of at least 2 in the proximal and mid-proximal coronary arteries of the LMCA, LAD, LCX, and RCA.

[0179] Example 8. Treatment of Kawasaki disease-related diseases using ROCK2 inhibitors and remdesivir The study will be conducted on 20 pediatric patients suspected of having SARS-CoV-2 infection or suspected of having SARS-CoV-2 infection and Kawasaki disease-related disease or pediatric multisystem inflammatory syndrome (PMIS) or pediatric inflammatory multisystem syndrome temporally associated with SARS-CoV-2 (PIMS-TS). Formula XII will be administered orally at 100-400 mg daily for a treatment period of 2-3 weeks. A prescribed amount of Formula XII will be administered at specified intervals. In parallel, remdesivir will be administered intravenously over 30-120 minutes in a total volume of 125 mL or less of 0.9% saline to pediatric patients weighing 3.5 kg to 40 kg, with a loading dose of 5 mg / kg on day 1 and a daily loading dose of 2.5 mg / kg on days 2-10.

[0180] Subjects are observed for approximately 2 weeks to approximately 6 months. Coronavirus testing is performed daily, every 2 days, every 3 days, or other intervals during the treatment period by collecting specimens from patients using a nasopharyngeal swab, oropharyngeal swab, mid-nasal turbinate swab, or anterior nares swab. The swab is then immediately placed into a sterile transport tube containing approximately 2-3 mL of either viral transport medium (VTM), Amies transport medium, or sterile saline, unless a coronavirus point-of-care test is used. Samples are tested using a CDC-approved testing method. Different doses of Formula XII 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 following the same protocol and the dose-response relationship for the degree of viral inhibition is recorded.

[0181] In addition to testing the extent of viral infection, vasculitis tests, such as blood tests (for C-reactive protein, complete blood count, amount of antineutrophil cytoplasmic antibodies, or other biomarkers), urinalysis (red blood cell or urinary protein content), and 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, 3, 4, or 5 days, weekly, biweekly, or monthly, to monitor the progression of Kawasaki-like disease, PMIS, or PIMS-TS. Furthermore, the period of administration of Formula XII can be extended beyond 2 weeks at different dose levels in this study.

[0182] To assess disease progression, patients undergo a 2-dimensional echocardiogram one month after the disease to evaluate coronary artery lesions (CAL). Measurements for each patient include the diameters of the left main coronary artery (LMCA), left anterior descending artery (LAD), left circumflex artery (LCX), and proximal and mid-right coronary artery (RCA). A Z-score is calculated 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 a z of at least 2 in the proximal and mid-proximal coronary arteries of the LMCA, LAD, LCX, and RCA.

Claims

1. 1. A pharmaceutical composition for treating a viral infection in a patient in need thereof, comprising a compound of formula I: 【Chemistry 1】 or a pharma- ceutical acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotopically labeled compound thereof, X and Y are each independently a direct bond, C(=O), O, or 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 ３～１０ Cyclic hydrocarbyl, saturated or partially unsaturated 3- to 10-membered heterocyclyl, C ６～１０ Aryl, 5-14 membered heteroaryl and C ６～１２ aralkyl, wherein up to two ring members of the cyclic hydrocarbyl and heterocyclyl are C(=O); Ring A and ring B each independently represent a saturated or partially unsaturated C ３～１０ Hydrocarbon ring, saturated or partially unsaturated 3- to 10-membered heterocyclic ring, C ６～１０ and 5-14 membered heteroaromatic rings, wherein up to two ring members of said hydrocarbon rings and heterocyclic rings are C(=O), with the proviso that when ring B is a heterocyclic ring containing a nitrogen atom, ring B is not bonded to X via said nitrogen atom; Ring C is a saturated or partially unsaturated C ３～１０ Hydrocarbon ring, saturated or partially unsaturated 3- to 10-membered heterocyclic ring, C ６～１０ selected from the group consisting of aromatic rings and 5-14 membered heteroaromatic rings, wherein up to two ring members of said hydrocarbon rings and heterocyclic rings are C(=O); Ring D is absent or is saturated or partially unsaturated C ３～１０ Hydrocarbon ring, saturated or partially unsaturated 3- to 10-membered heterocyclic ring, C ６～１０ selected from the group consisting of aromatic rings and 5-14 membered heteroaromatic rings, wherein up to two ring members of said hydrocarbon rings and heterocyclic rings are C(=O); Ring E is selected from the group consisting of: 【Chemistry 2】 Ring F is a saturated or partially unsaturated C ３～１０ Hydrocarbon ring, saturated or partially unsaturated 3- to 10-membered heterocyclic ring, C ６～１０ selected from the group consisting of aromatic rings and 5-14 membered heteroaromatic rings, wherein up to two ring members of said hydrocarbon rings and heterocyclic rings are C(=O); R １ is H, -NH ２ , C １～６ Alkyl, C ６～１０ aryl, 5- to 14-membered heteroaryl, N-methylpyrrolidinyl, N-methylpiperidinyl, 【Chemistry 3】 Acetyl, 【Chemistry 4】 -C(=O)-(C １～６ (alkylene) n-CF ３ , -C(=O)-(C １～６ alkylene)CN, -C(=O)-(saturated or partially unsaturated C ３～１０ Cyclic hydrocarbyl), -NHC(=O)-(saturated or partially unsaturated C ３～１０ cyclic hydrocarbyl), -C(=O)-(saturated or partially unsaturated 3- to 10-membered heterocyclyl), -C(=O)-C １～６ alkylene-(saturated or partially unsaturated 3- to 10-membered heterocyclyl), -C(=O)-(5- to 14-membered heteroaryl), -C(=O)-C １～６ Alkylene-NH(C １～６ alkyl), -C(=O)-C １～６ Alkylene-N(C １～６ Alkyl) ２ , N-methylpiperazine substituted acetyl, -S(=O) ２ R １ａ , -P(=O)R １ａ R １ｂ , 【Chemistry 5】 is selected from the group consisting of However, R １ and R １０ One of them is C １～６ alkyl and the other is H or C ３～１０ When it is a cyclic hydrocarbyl, at least one of X and Y is a direct bond, ring C is not a 5-membered heteroaromatic ring, and R １ and R １０ is H and the other is: 【Chemistry 6】 Ring C is not a 5-membered heteroaromatic ring, １ and R １０ are both H, ring A contains at least one nitrogen atom and is not a 5- or 6-membered ring, １ and R １０ is H and the other is: 【Chemistry 7】 Ring C is not a 5-membered heteroaromatic ring, １ and R １０ is H and the other is H or acetyl, then ring D is absent; R １ａ and R １ｂ are each independently H, halogen, amino, cyano, nitro, C １～６ Alkyl, C ２～６ Alkenyl, C ２～６ Alkynyl, C ３～１０ Cyclic hydrocarbyl, 3-10 membered heterocyclyl, C ６～１０ Aryl, 5-14 membered heteroaryl, C ６～１２ Aralkyl, -C(=O)R ５ , -OC(=O)R ５ , -C(=O)OR ５ , -OR ５ , -SR ５ , -S(=O)R ５ , -S(=O) ２ R ５ , -S(=O) ２ N.R. ５ R ６ , -NR ５ R ６ , -C(=O)NR ５ R ６ , -NR ５ -C(=O)R ６ , -NR ５ -C(=O)OR ６ , N.R. ５ -S(=O) ２ -R ６ , -NR ５ -C(=O)-NR ５ R ６ , -C １～６ Alkylene -NR ５ R ６ , -C １～６ Alkylene-OR ５ and -O-C １～６ Alkylene -NR ５ R ６ where R is selected from the group consisting of １ａ and R １ｂ is n-propyl, the other is not H, or R １ａ and R １ｂ form, together with the atoms to which they are attached, a 3- to 12-membered heterocyclic or heteroaromatic ring; R ２ , R ３ , R ４ , R ７ , R ８ , R ９ and R １０ is, at each occurrence, H, halogen, amino, cyano, nitro, C １～６ Alkyl, C ２～６ Alkenyl, C ２～６ Alkynyl, C ３～１０ Cyclic hydrocarbyl, 3-10 membered heterocyclyl, C ６～１０ Aryl, 5-14 membered heteroaryl, C ６～１２ Aralkyl, -C(=O)R ５ , OC(=O)R ５ , -C(=O)OR ５ , OR ５ , S.R. ５ , -S(=O)R ５ , -S(=O) ２ R ５ , -S(=O)2NR ５ R ６ , -NR ５ R ６ , -C(=O)NR ５ R ６ , -NR ５ -C(=O)R ６ , -NR ５ -C(=O)OR ６ , -NR ５ -S(=O) ２ -R ６ , -NR ５ -C(=O)-NR ５ R ６ , -C １～６ Alkylene -NR ５ R ６ , -C １～６ Alkylene-O(P=O)(OH) ２ and -O-C １～６ Alkylene -NR ５ R ６ are each independently selected from the group consisting of: The above alkyl, alkylene, alkenyl, alkynyl, cyclic hydrocarbyl, hydrocarbon ring, heterocyclyl, heterocycle, aryl, aromatic ring, heteroaryl, heteroaromatic ring and aralkyl are each independently selected from the group consisting of halogen, hydroxyl, oxo, amino, cyano, nitro, C １～６ Alkyl, C ２～６ Alkenyl, C ２～６ Alkynyl, C ３～６ Cyclic hydrocarbyl, 3-10 membered heterocyclyl, C ６～１０ Aryl, 5-14 membered heteroaryl, C ６～１２ Aralkyl, =N-OR ５ , -C(=NH)NH ２ , -C(=O)R ５ , -OC(=O)R ５ , -C(=O)OR ５ , -OR ５ , -SR ５ , -S(=O)R ５ , -S(=O) ２ R ５ , -S(=O) ２ N.R. ５ R ６ , -NR ５ R ６ , -C(=O)NR ５ R ６ , -NR ５ -C(=O)R ６ , -NR ５ -C(=O)OR ６ , -NR ５ -S(=O) ２ -R ６ , -NR ５ -C(=O)-NR ５ R ６ , -C １～６ Alkylene -NR ５ R ６ and -O-C １～６ Alkylene -NR ５ R ６ wherein the alkyl, cyclic hydrocarbyl, heterocyclyl, aryl, heteroaryl and aralkyl are each optionally substituted with one or more substituents independently selected from the group consisting of halogen, hydroxyl, oxo, amino, cyano, nitro, C １～６ Alkyl, C ３～６ Cyclic hydrocarbyl, 3-10 membered heterocyclyl, C ６～１０ Aryl, 5-14 membered heteroaryl and C ６～１２ aryl, arylalkyl, R ５ and R ６ is H, alkyl, C ３～１０ Cyclic hydrocarbyl, 3-10 membered heterocyclyl, C ６～１０ Aryl, 5-14 membered heteroaryl and C ６～１２ aralkyl, m, in each occurrence, is independently an integer of 0, 1, 2, or 3; n is an integer of 0, 1, or 2; i is an integer of 0, 1, or 2; and g is an integer of 0, 1, 2, 3, or 4.

2. The ROCK2 inhibitor has the formula II-IX: 【Chemistry 8-1】 【Chemistry 8-2】 【Chemistry 8-3】 or a pharma- ceutical acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotopically labeled compound thereof, 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 above.

3. The ROCK2 inhibitor has formula X or formula XI: 【Chemistry 9】 or a pharma- ceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotopically labeled compound thereof, wherein R is H and C １～６ is selected from the group consisting of alkyl, Ring D is a saturated or partially unsaturated 3- to 10-membered heterocycle, C ６～１０ aryl, or a 5- to 10-membered heteroaromatic ring, preferably 【Chemistry 10】 a phenyl ring, an N-methylpyrrole ring, a furan ring, or a thiophene ring; R ２ is H and C １～６ is selected from the group consisting of alkyl, R ３ , R ４ , R ７ , R ７’ and R ８ is H, halogen, -NH at each occurrence. ２ , -OH, C １～６ Alkyl and -OR ５ are each independently selected from the group consisting of: R ９ and R １０ are H, halogen, and C at each occurrence. １～６ Alkyl, C ２～６ Alkenyl, C ３～１０ Cyclic hydrocarbyl, 3-10 membered heterocyclyl, C ６～１０ Aryl, 5-14 membered heteroaryl, C ６～１２ Aralkyl, -C(=O)R ５ , and -C １～６ Alkylene-O(P=O)(OH) ２ are each independently selected from the group consisting of: The above alkyl, alkenyl, cyclic hydrocarbyl, heterocyclyl, aryl, heteroaryl, heteroaromatic ring and aralkyl are each independently selected from the group consisting of halogen, C １～６ Alkyl and -OR ５ each optionally substituted with one or more substituents independently selected from the group consisting of: R ５ and R ６ In each occurrence, H, C １～６ Alkyl, C ３～１０ Cyclic hydrocarbyl, 3-10 membered heterocyclyl, C ６～１０ Aryl, 5-14 membered heteroaryl and C ６～１２ aralkyl, m, in each occurrence, is independently an integer of 0, 1, 2, or 3; and n is an integer of 0, 1, or 2.

4. R ５ and R ６ The pharmaceutical composition of any one of claims 1 to 3, wherein each occurrence is independently selected from the group consisting of H, methyl and ethyl.

5. R ３ , R ４ , R ７ , R ７’ and R ８ are H, F, Cl, Br, -NH ２ , —OH, methyl, trifluoromethyl, —CH ２ -Ph, methoxy, ethoxy and -CH ２ OCH ３ The pharmaceutical composition according to any one of claims 1 to 4, each independently selected from the group consisting of:

6. R ９ and R １０ is each occurrence H, F, Cl, Br, methyl, ethyl, n-propyl, isopropyl, vinyl, cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl, monofluoromethyl, difluoromethyl, trifluoromethyl, acetyl, -OCH ２ CHF ２ , C.H. ２ OH, -CH ２ OCH ３ , -CH ２ CH ２ OCH ３ , -CH ２ -O(P=O)(OH) ２ , 【Chemistry 11】 The pharmaceutical composition according to any one of claims 1 to 5, each independently selected from the group consisting of:

7. The ROCK2 inhibitor has formula XII: 【Chemistry 12】 2. The pharmaceutical composition of claim 1, which is the compound (6-(4-((4-(1H-pyrazol-4-yl)phenyl)amino)pyrimidin-2-yl)-1-methyl-1H-indol-2-yl)(3,3-difluoroazetidin-1-yl)methanone having the formula:

8. The pharmaceutical composition according to any one of claims 1 to 7, wherein the viral infection is caused by a coronavirus.

9. 1. A pharmaceutical composition for treating or preventing sequelae of a viral infection in a patient in need thereof, comprising a compound of formula I: 【Chemistry 13】 or a pharma- ceutical acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotopically labeled compound thereof, X and Y are each independently a direct bond, C(=O), O, or 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 ３～１０ Cyclic hydrocarbyl, saturated or partially unsaturated 3- to 10-membered heterocyclyl, C ６～１０ Aryl, 5-14 membered heteroaryl and C ６～１２ aralkyl, wherein up to two ring members of the cyclic hydrocarbyl and heterocyclyl are C(=O); Ring A and ring B each independently represent a saturated or partially unsaturated C ３～１０ Hydrocarbon ring, saturated or partially unsaturated 3- to 10-membered heterocyclic ring, C ６～１０ and 5-14 membered heteroaromatic rings, wherein up to two ring members of said hydrocarbon rings and heterocyclic rings are C(=O), with the proviso that when ring B is a heterocyclic ring containing a nitrogen atom, ring B is not bonded to X via said nitrogen atom; Ring C is a saturated or partially unsaturated C ３～１０ Hydrocarbon ring, saturated or partially unsaturated 3- to 10-membered heterocyclic ring, C ６～１０ selected from the group consisting of aromatic rings and 5-14 membered heteroaromatic rings, wherein up to two ring members of said hydrocarbon rings and heterocyclic rings are C(=O); Ring D is absent or is saturated or partially unsaturated C ３～１０ Hydrocarbon ring, saturated or partially unsaturated 3- to 10-membered heterocyclic ring, C ６～１０ selected from the group consisting of aromatic rings and 5-14 membered heteroaromatic rings, wherein up to two ring members of said hydrocarbon rings and heterocyclic rings are C(=O); Ring E is selected from the group consisting of: 【Chemistry 14】 Ring F is a saturated or partially unsaturated C ３～１０ Hydrocarbon ring, saturated or partially unsaturated 3- to 10-membered heterocyclic ring, C ６～１０ selected from the group consisting of aromatic rings and 5-14 membered heteroaromatic rings, wherein up to two ring members of said hydrocarbon rings and heterocyclic rings are C(=O); R １ is H, -NH ２ , C １～６ Alkyl, C ６～１０ aryl, 5- to 14-membered heteroaryl, N-methylpyrrolidinyl, N-methylpiperidinyl, 【Chemistry 15】 Acetyl, 【Chemistry 16】 -C(=O)-(C １～６ (alkylene) n-CF ３ , -C(=O)-(C １～６ alkylene)CN, -C(=O)-(saturated or partially unsaturated C ３～１０ Cyclic hydrocarbyl), -NHC(=O)-(saturated or partially unsaturated C ３～１０ cyclic hydrocarbyl), -C(=O)-(saturated or partially unsaturated 3- to 10-membered heterocyclyl), -C(=O)-C １～６ alkylene-(saturated or partially unsaturated 3- to 10-membered heterocyclyl), -C(=O)-(5- to 14-membered heteroaryl), -C(=O)-C １～６ Alkylene-NH(C １～６ alkyl), -C(=O)-C １～６ Alkylene-N(C １～６ Alkyl) ２ , N-methylpiperazine substituted acetyl, -S(=O) ２ R １ａ , -P(=O)R １ａ R １ｂ , 【Chemistry 17】 is selected from the group consisting of However, R １ and R １０ One of them is C １～６ alkyl and the other is H or C ３～１０ When it is a cyclic hydrocarbyl, at least one of X and Y is a direct bond, ring C is not a 5-membered heteroaromatic ring, and R １ and R １０ is H and the other is: 【Chemistry 18】 Ring C is not a 5-membered heteroaromatic ring, １ and R １０ are both H, ring A contains at least one nitrogen atom and is not a 5- or 6-membered ring, １ and R １０ is H and the other is: 【Chemistry 19】 Ring C is not a 5-membered heteroaromatic ring, １ and R １０ is H and the other is H or acetyl, then ring D is absent; R １ａ and R １ｂ are each independently H, halogen, amino, cyano, nitro, C １～６ Alkyl, C ２～６ Alkenyl, C ２～６ Alkynyl, C ３～１０ Cyclic hydrocarbyl, 3-10 membered heterocyclyl, C ６～１０ Aryl, 5-14 membered heteroaryl, C ６～１２ Aralkyl, -C(=O)R ５ , -OC(=O)R ５ , -C(=O)OR ５ , -OR ５ , -SR ５ , -S(=O)R ５ , -S(=O) ２ R ５ , -S(=O) ２ N.R. ５ R ６ , -NR ５ R ６ , -C(=O)NR ５ R ６ , -NR ５ -C(=O)R ６ , -NR ５ -C(=O)OR ６ , N.R. ５ -S(=O) ２ -R ６ , -NR ５ -C(=O)-NR ５ R ６ , -C １～６ Alkylene -NR ５ R ６ , -C １～６ Alkylene-OR ５ and -O-C １～６ Alkylene -NR ５ R ６ where R is selected from the group consisting of １ａ and R １ｂ is n-propyl, the other is not H, or R １ａ and R １ｂ form, together with the atoms to which they are attached, a 3- to 12-membered heterocyclic or heteroaromatic ring; R ２ , R ３ , R ４ , R ７ , R ８ , R ９ and R １０ is, at each occurrence, H, halogen, amino, cyano, nitro, C １～６ Alkyl, C ２～６ Alkenyl, C ２～６ Alkynyl, C ３～１０ Cyclic hydrocarbyl, 3-10 membered heterocyclyl, C ６～１０ Aryl, 5-14 membered heteroaryl, C ６～１２ Aralkyl, -C(=O)R ５ , OC(=O)R ５ , -C(=O)OR ５ , OR ５ , S.R. ５ , -S(=O)R ５ , -S(=O) ２ R ５ , -S(=O)2NR ５ R ６ , -NR ５ R ６ , -C(=O)NR ５ R ６ , -NR ５ -C(=O)R ６ , -NR ５ -C(=O)OR ６ , -NR ５ -S(=O) ２ -R ６ , -NR ５ -C(=O)-NR ５ R ６ , -C １～６ Alkylene -NR ５ R ６ , -C １～６ Alkylene-O(P=O)(OH) ２ and -O-C １～６ Alkylene -NR ５ R ６ are each independently selected from the group consisting of: The above alkyl, alkylene, alkenyl, alkynyl, cyclic hydrocarbyl, hydrocarbon ring, heterocyclyl, heterocycle, aryl, aromatic ring, heteroaryl, heteroaromatic ring and aralkyl are each independently selected from the group consisting of halogen, hydroxyl, oxo, amino, cyano, nitro, C １～６ Alkyl, C ２～６ Alkenyl, C ２～６ Alkynyl, C ３～６ Cyclic hydrocarbyl, 3-10 membered heterocyclyl, C ６～１０ Aryl, 5-14 membered heteroaryl, C ６～１２ Aralkyl, =N-OR ５ , -C(=NH)NH ２ , -C(=O)R ５ , -OC(=O)R ５ , -C(=O)OR ５ , -OR ５ , -SR ５ , -S(=O)R ５ , -S(=O) ２ R ５ , -S(=O) ２ N.R. ５ R ６ , -NR ５ R ６ , -C(=O)NR ５ R ６ , -NR ５ -C(=O)R ６ , -NR ５ -C(=O)OR ６ , -NR ５ -S(=O) ２ -R ６ , -NR ５ -C(=O)-NR ５ R ６ , -C １～６ Alkylene -NR ５ R ６ and -O-C １～６ Alkylene -NR ５ R ６ wherein the alkyl, cyclic hydrocarbyl, heterocyclyl, aryl, heteroaryl and aralkyl are each optionally substituted with one or more substituents independently selected from the group consisting of halogen, hydroxyl, oxo, amino, cyano, nitro, C １～６ Alkyl, C ３～６ Cyclic hydrocarbyl, 3-10 membered heterocyclyl, C ６～１０ Aryl, 5-14 membered heteroaryl and C ６～１２ aryl, arylalkyl, R ５ and R ６ is H, alkyl, C ３～１０ Cyclic hydrocarbyl, 3-10 membered heterocyclyl, C ６～１０ Aryl, 5-14 membered heteroaryl and C ６～１２ aralkyl, m, in each occurrence, is independently an integer of 0, 1, 2, or 3; n is an integer of 0, 1, or 2; i is an integer of 0, 1, or 2; and g is an integer of 0, 1, 2, 3, or 4.

10. The ROCK2 inhibitor has the formula II-IX: 【Chemistry 20-1】 【Chemistry 20-2】 【Chemistry 20-3】 or a pharma- ceutical acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotopically labeled compound thereof, 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 above.

11. The ROCK2 inhibitor has formula X or formula XI: 【Chemistry 21】 or a pharma- ceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, or isotopically labeled compound thereof, wherein R is H and C １～６ is selected from the group consisting of alkyl, Ring D is a saturated or partially unsaturated 3- to 10-membered heterocycle, C ６～１０ aryl, or a 5- to 10-membered heteroaromatic ring, preferably 【Chemical 22】 a phenyl ring, an N-methylpyrrole ring, a furan ring, or a thiophene ring; R ２ is H and C １～６ is selected from the group consisting of alkyl, R ３ , R ４ , R ７ , R ７’ and R ８ is H, halogen, -NH at each occurrence. ２ , -OH, C １～６ Alkyl and -OR ５ are each independently selected from the group consisting of: R ９ and R １０ are H, halogen, and C at each occurrence. １～６ Alkyl, C ２～６ Alkenyl, C ３～１０ Cyclic hydrocarbyl, 3-10 membered heterocyclyl, C ６～１０ Aryl, 5-14 membered heteroaryl, C ６～１２ Aralkyl, -C(=O)R ５ , and -C １～６ Alkylene-O(P=O)(OH) ２ are each independently selected from the group consisting of: The above alkyl, alkenyl, cyclic hydrocarbyl, heterocyclyl, aryl, heteroaryl, heteroaromatic ring and aralkyl are each independently selected from the group consisting of halogen, C １～６ Alkyl and -OR ５ each optionally substituted with one or more substituents independently selected from the group consisting of: R ５ and R ６ In each occurrence, H, C １～６ Alkyl, C ３～１０ Cyclic hydrocarbyl, 3-10 membered heterocyclyl, C ６～１０ Aryl, 5-14 membered heteroaryl and C ６～１２ aralkyl, m, in each occurrence, is independently an integer of 0, 1, 2, or 3; and n is an integer of 0, 1, or 2.

12. R ５ and R ６ The pharmaceutical composition of any one of claims 9 to 11, wherein each occurrence is independently selected from the group consisting of H, methyl and ethyl.

13. R ３ , R ４ , R ７ , R ７’ and R ８ are H, F, Cl, Br, -NH ２ , —OH, methyl, trifluoromethyl, —CH ２ -Ph, methoxy, ethoxy and -CH ２ OCH ３ The pharmaceutical composition according to any one of claims 9 to 12, each independently selected from the group consisting of:

14. R ９ and R １０ is each occurrence H, F, Cl, Br, methyl, ethyl, n-propyl, isopropyl, vinyl, cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl, monofluoromethyl, difluoromethyl, trifluoromethyl, acetyl, -OCH ２ CHF ２ , C.H. ２ OH, -CH ２ OCH ３ , -CH ２ CH ２ OCH ３ , -CH ２ -O(P=O)(OH) ２ 【Chemistry 23】 The pharmaceutical composition according to any one of claims 9 to 13, each independently selected from the group consisting of:

15. The ROCK2 inhibitor has formula XII: 【Chemistry 24】 10. The pharmaceutical composition of claim 9, which is the compound (6-(4-((4-(1H-pyrazol-4-yl)phenyl)amino)pyrimidin-2-yl)-1-methyl-1H-indol-2-yl)(3,3-difluoroazetidin-1-yl)methanone having the formula:

16. The pharmaceutical composition according to any one of claims 1 to 15, wherein the viral infection is caused by a coronavirus.

17. 17. The pharmaceutical composition of claim 16, wherein the viral infection is caused by SARS-CoV-1, SARS-CoV-2, or MERS-CoV.

18. The pharmaceutical composition of claim 17, wherein the viral infection is caused by SARS-CoV-2.

19. The pharmaceutical composition of claim 18, wherein the viral infection is caused by SARS-CoV-2 variant delta and / or SARS-CoV-2 variant omicron.

20. 20. The pharmaceutical composition of any one of claims 9 to 19, wherein the sequelae of the viral infection include one or more of the group consisting of fatigue, dyspnea, cough, joint pain, muscle pain, headache, chest pain, fever, palpitations, myocardial inflammation, ventricular dysfunction, stroke, pulmonary function abnormalities, pulmonary fibrosis, renal dysfunction rash, alopecia, smell and / or taste disorders, sleep regulation disorders, cognitive impairment changes, memory impairment, depression, anxiety, mood changes and combinations thereof.

21. The pharmaceutical composition according to any one of claims 9 to 19, wherein the sequelae are fibrosis.

22. 22. The pharmaceutical composition of any one of claims 1 to 21, wherein said ROCK2 inhibitor is administered to said patient in a total dosage of about 200 mg to about 500 mg per day.

23. 23. The pharmaceutical composition of claim 22, wherein the ROCK2 inhibitor is administered to the patient in a total dose of about 200 mg per day.

24. 23. The pharmaceutical composition of claim 22, wherein the ROCK2 inhibitor is administered to the patient in a total dose of about 300 mg per day.

25. 23. The pharmaceutical composition of claim 22, wherein the ROCK2 inhibitor is administered to the patient in a total dose of about 400 mg per day.

26. 23. The pharmaceutical composition of claim 22, wherein the ROCK2 inhibitor is administered to the patient in a total dose of about 500 mg per day.

27. The pharmaceutical composition of any one of claims 22 to 26, which is administered once a day.

28. The pharmaceutical composition of any one of claims 22 to 26, administered twice daily.

29. The pharmaceutical composition of any one of claims 1 to 28, administered to a patient within about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 24, about 28, about 72, about 96, or about 120 hours after the patient is first exposed to the virus that causes the viral infection.

30. The pharmaceutical composition of any one of claims 1 to 28, which is administered to the patient within about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 24, about 28, about 72, about 96, or about 120 hours after the patient develops symptoms caused by the viral infection.

31. The pharmaceutical composition of claim 30, administered to the patient for about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, or about 10 weeks after the symptoms have subsided.

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