Composition for inhibiting herpes virus

Abstract

The present disclosure provides compounds of Formula I-V that are useful for inhibiting a viral disease in a subject. In some embodiments, the compounds of Formula I-V may be used to inhibit herpes virus.

Description

[0001] Cross - Reference to Related Applications

[0002] This application claims the benefit of U.S. Application No. 62 / 965,755, filed on January 24, 2020, the disclosure of which is incorporated herein by reference in its entirety.

[0003] Sequence Listing

[0004] This application contains a Sequence Listing submitted electronically in ASCII format and incorporated herein by reference in its entirety. The ASCII copy, created on January 22, 2021, is named 2635 - 000IWO0I_Sequence Listing_ST25.txt and is 1 KB in size.

[0005] Statement of Government Interests

[0006] This invention was made with government support under Grant No. 1R0IDC013550 awarded by the National Institutes of Health. The government has certain rights in this invention. Background of the Invention

[0008] Infection with human cytomegalovirus (HCMV), which is a member of the herpes virus family, is common in humans. The seropositivity rate reaches 90% in individuals over 80 years old. 1 Although the infection is usually asymptomatic, HCMV still poses a serious threat to transplant recipients and AIDS patients. It is also the most common congenital infection worldwide, causing hearing loss, global disabilities, and central nervous system damage in children. 2-6 .

[0009] The nucleoside analogue ganciclovir (GCV) and its oral formulation val - GCV have significantly improved transplant outcomes by reducing HCMV replication and sequelae of the disease. 7,8 GCV has also been shown to prevent hearing deterioration in congenitally infected children. 9 However, prolonged courses of GCV or val - GCV lead to severe myelotoxicity. 10-13 A phase III clinical trial of oral val - GCV in congenitally infected infants concluded that 6 - month treatment may have better neurological effects than 6 weeks, but GCV - resistant mutants emerged. 14 Similarly, GCV - resistant mutants have emerged in transplant recipients. 15-16When ganciclovir resistance occurs, the limited treatment alternatives, foscarnet and cidofovir, are highly nephrotoxic and can only be administered intravenously. Their use in resistant or refractory HCMV cases is associated with high morbidity and mortality. 13 Some new drugs have entered FDA approval and clinical trials. The terminase inhibitor letermovir was recently approved for HCMV prophylaxis after hematopoietic stem cell transplantation. 17 Rapid selection of UL56 mutations in cell culture has been reported. 16 and patient resistance to letermovir. 18 The viral UL97 kinase inhibitor maribavir is in clinical trials. 19

[0010] The identification of new herpesvirus inhibitors is an important area of drug development. Despite the availability of multiple drugs, there is clearly a need to improve existing drugs given the toxicity of the available drugs and the ongoing selection of drug-resistant mutants. Summary of the Invention

[0012] Disclosed herein are methods of inhibiting human herpesvirus in a subject in need thereof, the methods comprising administering to the subject a compound of formula I or a pharmaceutical composition thereof or a salt, solvate or stereoisomer thereof, the pharmaceutical composition comprising a pharmaceutically acceptable carrier:

[0013]

[0014] wherein R 1 is a cycloalkyl, heterocycloalkyl, aryl or heteroaryl optionally substituted with one or more alkyl, amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidino, acyl guanidino, having 1 to 10 carbon atoms and linked by a C 1 -C 3 alkyl linkage. In some embodiments, the compound is selected from the group consisting of:

[0015]

[0016]

[0017] Disclosed herein are methods of inhibiting human herpesvirus in a subject in need thereof, the methods comprising administering to the subject a compound of formula II or a pharmaceutical composition thereof or a salt, solvate or stereoisomer thereof, the pharmaceutical composition comprising a pharmaceutically acceptable carrier:

[0018] ​

[0019] Wherein, R 1 is H, and R 2 is independently H or an alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl optionally substituted by one or more alkyl, amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidine, acyl guanidine, having 1 to 10 carbon atoms and connected through C 1 -C 3 alkyl; or R 1 and R 2 together form a C 1 -C 6 heterocycloalkyl or C 1 -C 6 heteroaryl optionally substituted by one or more amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidine, acyl guanidine, having 1 to 10 carbon atoms; and R 3 is H or a cycloalkyl, heterocycloalkyl, aryl or heteroaryl optionally substituted by one or more halogen, CF 3 , CN and NO 2 moieties. In some embodiments, R 2 is a C 1 -C 6 heterocycloalkyl or C 1 -C 6 heteroaryl optionally substituted by one or more alkyl, amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidine, acyl guanidine, having 1 to 10 carbon atoms and connected through C 1 -C 3 alkyl. In some embodiments, the compound is selected from the group consisting of:

[0020]

[0021] Disclosed herein is a method for inhibiting human herpesvirus in a subject in need thereof, the method comprising administering to the subject a compound of formula III or a pharmaceutical composition thereof or a salt, solvate or stereoisomer thereof, the pharmaceutical composition comprising a pharmaceutically acceptable carrier:

[0022]

[0023] Disclosed herein is a method for inhibiting human herpesvirus in a subject in need thereof, the method comprising administering a compound of formula IV or a pharmaceutical composition thereof or a salt, solvate or stereoisomer thereof, the pharmaceutical composition comprising a pharmaceutically acceptable carrier:

[0024]

[0025] The present disclosure provides a method for inhibiting human herpesvirus in a subject in need thereof, the method comprising administering to the subject a compound of formula V or a pharmaceutical composition thereof or a salt, solvate or stereoisomer thereof, the pharmaceutical composition comprising a pharmaceutically acceptable carrier:

[0026]

[0027] wherein R 1 is hydrogen, a C1-C6 straight or branched alkyl or a C3-C6 cycloalkyl; and R 2 is an unsubstituted or R 3 or OR 3 substituted phenyl, wherein R 3 is hydrogen, a C1-C6 straight or branched alkyl or a C3-C6 cycloalkyl. In some embodiments, R 1 is a C1-C4 straight or branched alkyl; and R 2 is a phenyl substituted by OR 3 wherein R 3 is a C1-C4 straight or branched alkyl. In some embodiments, the compound is

[0028]

[0029] In some embodiments, the methods disclosed herein further comprise administering more than one bioactive agent, e.g., two, three, four or more bioactive agents.

[0030] In some embodiments, the herpesvirus is selected from human herpesvirus type 1 (HHV-1), human herpesvirus type 2 (HHV2), human herpesvirus type 3 (HHV3), human herpesvirus type 4 (HHV4), human herpesvirus type 5 (HHV5), human herpesvirus type 6 (HHV6), human herpesvirus type 7 (HHV7) and human herpesvirus type 8 (HHV8). In some embodiments, the herpesvirus is HHV-5.

[0031] The present disclosure provides a pharmaceutical composition comprising a compound of formula I or a salt, solvate or stereoisomer thereof:

[0032]

[0033] wherein R 1 is a cycloalkyl, heterocycloalkyl, aryl or heteroaryl optionally substituted by one or more amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidine, acyl guanidine, having 1 to 10 carbon atoms and attached via C1 -C 3 Alkyl linkage. In some embodiments, the compound is selected from the group consisting of:

[0034]

[0035]

[0036] Disclosed herein is a pharmaceutical composition comprising a compound of formula II or a salt, solvate or stereoisomer thereof:

[0037]

[0038] Wherein, R 1 is H, and R 2 is independently H or an alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl optionally substituted by one or more alkyl, amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidine, acyl guanidine, having 1 to 10 carbon atoms and linked through C 1 -C 3 alkyl linkage; or R 1 and R 2 together form a C 1 -C 6 heterocycloalkyl or C 1 -C 6 heteroaryl optionally substituted by one or more amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidine, acyl guanidine, having 1 to 10 carbon atoms; and R 3 is H or a cycloalkyl, heterocycloalkyl, aryl or heteroaryl optionally substituted by one or more halogen, CF 3 , CN and NO 2 moieties. In some embodiments, R 2 is a C 1 -C 6 heterocycloalkyl or C 1 -C 6 heteroaryl optionally substituted by one or more alkyl, amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidine, acyl guanidine, having 1 to 10 carbon atoms and linked through C 1 -C 3 alkyl linkage. In some embodiments, the compound is selected from the group consisting of:

[0039]

[0040] The present disclosure relates to a pharmaceutical composition comprising a compound of formula III or a salt, solvate or stereoisomer thereof, and a pharmaceutically acceptable carrier:

[0041]

[0042] The present disclosure relates to a pharmaceutical composition comprising a compound of formula IV or a salt, solvate or stereoisomer thereof, and a pharmaceutically acceptable carrier:

[0043]

[0044] The present disclosure relates to a pharmaceutical composition comprising a compound of formula V or a salt, solvate or stereoisomer thereof, and a pharmaceutically acceptable carrier:

[0045]

[0046] wherein R 1 is hydrogen, a C1-C6 straight or branched alkyl, or a C3-C6 cycloalkyl; and R 2 is an unsubstituted or R 3 - or OR 3 -substituted phenyl, wherein R 3 is hydrogen, a C1-C6 straight or branched alkyl, or a C3-C6 cycloalkyl. In some embodiments, R 1 is a C1-C4 straight or branched alkyl; and R 2 is a phenyl substituted with OR 3 wherein R 3 is a C1-C4 straight or branched alkyl. In some embodiments, the compound is

[0047]

[0048] The present disclosure relates to a compound of formula I or a salt, solvate or stereoisomer thereof:

[0049]

[0050] wherein R 1 is a cycloalkyl, heterocycloalkyl, aryl or heteroaryl optionally substituted with one or more amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidine, acyl guanidine, having 1 to 10 carbon atoms and linked through a C 1 -C 3 alkyl, provided that the compound cannot be compound A:

[0051]

[0052] In some embodiments of the compound or its salt, solvate or stereoisomer, the compound is selected from:

[0053]

[0054] Disclosed herein are compounds of formula II or their salts, solvates or stereoisomers:

[0055]

[0056] Wherein, R 1 is H, and R 2 is independently H or an alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl optionally substituted by one or more alkyl, amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidine, acyl guanidine, having 1 to 10 carbon atoms and connected by C 1 -C 3 alkyl; or R 1 and R 2 together form a C 1 -C 6 heterocycloalkyl or C 1 -C 6 heteroaryl, optionally substituted by one or more amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidine, acyl guanidine, having 1 to 10 carbon atoms; and R 3 is H or a cycloalkyl, heterocycloalkyl, aryl or heteroaryl optionally substituted by one or more halogen, CF 3 , CN and NO 2 moieties, provided that the compound cannot be a compound selected from the group consisting of:

[0057]

[0058] In some embodiments, R 2 is a C 1 -C 6 heterocycloalkyl or C 1 -C 6 heteroaryl, optionally substituted by one or more alkyl, amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidine, acyl guanidine, having 1 to 10 carbon atoms and connected by C 1 -C 3 alkyl.

[0059] Disclosed herein are compounds of formula V or their salts, solvates or stereoisomers:

[0060]

[0061] Wherein, R 1 is hydrogen, a C1-C6 straight or branched alkyl group, or a C3-C6 cycloalkyl group; and R 2 is an unsubstituted or R 3 or OR 3 substituted phenyl group, wherein R 3 is hydrogen, a C1-C6 straight or branched alkyl group, or a C3-C6 cycloalkyl group. In some embodiments, R 1 is a C1-C4 straight or branched alkyl group; and R 2 is a phenyl group substituted by OR 3 wherein R 3 is a C1-C4 straight or branched alkyl group. In some embodiments, the compound is

[0062]

[0063] Disclosed herein is a method for inhibiting or treating human herpesvirus in a subject, the method comprising: administering to the subject a therapeutically effective amount of a pharmaceutically acceptable composition, the composition comprising one or more of the above compounds and a pharmaceutically acceptable carrier.

[0064] Also disclosed herein is a method for inhibiting or treating human herpesvirus in a subject, the method comprising: administering to the subject a therapeutically effective amount of a pharmaceutically acceptable composition, the composition comprising one or more of the above compounds, at least one other bioactive agent, and a pharmaceutically acceptable carrier.

[0065] Also disclosed herein is the use of the compounds or compositions disclosed herein for treating human herpesvirus in a subject. Also disclosed herein is the use of the compounds or compositions disclosed herein for treating human herpesvirus in a subject. Also disclosed herein is the use of the compounds or compositions disclosed herein for manufacturing a medicament for treating human herpesvirus in a subject. Brief Description of the Drawings

[0067] Figure 1 : Flow chart of high-throughput screening (HTS).

[0068] Figure 2 : Chemical structures and molecular weights (MW) of five compounds.

[0069] Figure 3 : MLS8969 inhibits HCMV replication, Figure 3 A: HFF was infected with HCMV Towne pp28 (MOI = 1), and then treated with MLS8999 or GCV. Luciferase activity was measured at 72 hpi. Figure 3B: HFFs were infected with HCMV TB40 (100 PFU / well) and treated with MLS8969 or GCV. Plaques were stained and counted after 10 days. Figure 3 C: HFFs were pretreated with 8969 or GCV for 24 h and then infected with HCMV Towne at MOIs of 1, 0.1, and 0.01. Luciferase activity was measured at 72 hpi. Figure 3 D, Figure 3 E: HFFs were pretreated with the indicated concentrations of MLS8969 or GCV for 24 h and then infected with HCMV TB40 ( Figure 3 D) or GCV-resistant HCMV Towne ( Figure 3 E) at 100 PFU / well. Plaques were stained and counted after 10 days. Figure 3 A-3E: Data are the mean ± SD of three values from a representative experiment. Figure 3 F: HFFs were pretreated or infected and then treated. Infections were performed at MOIs of 1, 0.1, and 0.01 (Towne). Expression of viral proteins from cell lysates was measured at 72 hpi. WB data are from a representative experiment.

[0070] Figure 4 : MLS8969 is an entry inhibitor of HCMV. HFFs were pretreated with 3 μM MLS8999 for 24 h and then infected with HCMV Towne (MOI = 1, 0.1, 0.01). Heparin (30 μM) was used as a positive control to block virus entry into cells, and GCV pretreatment (5 μM) was used as a negative control. IFA for HCMV-encoded pp65 was performed at 2 hpi to determine the amount of pp65 localized in the nucleus under different conditions. The experiment was independently repeated three times, and images from a representative experiment are shown.

[0071] Figure 5 : NFU1827 is an immediate early-early inhibitor of HCMV replication. Figure 5 A: HFFs were infected with HCMV TB40 (100 PFU / cell) and treated with the indicated concentrations of the compound NFU1827. Plaques were stained and counted at 8 dpi. The experiment was repeated three times, and the data shown are the mean ± SD of three values from a representative experiment. Figure 5 B: HFFs were infected with GCV-resistant HCMV Towne (MOI = 0.1 PFU / cell) and treated with the indicated concentrations of NFU1827. Luciferase activity in cell lysates was measured at 72 hpi. The data shown are the mean ± SD of three values from a representative experiment. Figure 5C, 5D: HFFs were infected with HCMV Towne (MOI = 1 PFU / cell), and compounds were added or removed after infection (0, 6, 12, 24, 48, and 72 h). Luciferase activity in cell lysates was measured at 72 hpi. The data shown are the mean ± SD of three values from a representative experiment. Figure 5 E: HFFs were infected with HCMV Towne (MOI = 1 PFU / cell), and the expression of viral proteins and cellular β-actin was determined at 72 hpi. The experiment was repeated twice, and the WB data are from a representative experiment. Figure 5 F, 5G: HFFs were infected with HCMV TB40 (MOI = 0.1) and treated with NFU1827 (5 μM) or GCV (5 μM). Cells were lysed at the indicated time points to isolate DNA. Viral DNA replication in cells ( Figure 5 F) and viral DNA load in the supernatant ( Figure 5 G) were measured by real-time PCR. The experiment was repeated three times, and the data shown are the mean ± SD of triplicate values from a representative experiment.

[0072] Figure 6 : MLS8554 is an immediate-early inhibitor of HCMV replication. Figure 6 A: HFFs were infected with TB40 at 100 PFU / well and treated with the indicated concentrations of MLS8554. Plaques were stained and counted at 8 dpi. The experiment was repeated three times, and the data shown are from a representative experiment. Figure 6 B: HFFs were infected with GCV-resistant HCMV Towne (MOI = 0.1) and treated with the indicated concentrations of MLS8554. Luciferase activity in cell lysates was measured at 72 hpi. The data shown are the mean ± SD of three values from a single experiment. Figure 6 C, 6D: HFFs were infected with HCMV Towne (MOI = 1), and compounds were added or removed at the indicated time points after infection. Luciferase activity in cell lysates was measured at 72 hpi. The experiment was independently repeated twice, and the representative data from a single experiment are shown. Figure 6 E: HFFs were infected with HCMV Towne (MOI = 1), and the expression of viral proteins and cellular β-actin was determined 3 days later. The WB data are from a representative experiment. Figure 6 F, 6G: HFFs were infected with HCMV Towne (MOI = 0.1) and treated with MLS8554 (2 μM) or GCV (5 μM). Supernatants were collected, and cells were lysed at the indicated time points to isolate DNA. Viral DNA replication in cells ( Figure 6 F) and viral DNA load in the supernatant ( Figure 6G). The data shown are the mean ± SD of four values from two independent experiments.

[0073] Figure 7 A: MLS8091 inhibits HCMV replication in the early and late stages. Figure 7 A: HFFs were infected with HCMV TB40 (100 PFU / well) and treated with the indicated concentrations of MLS8091. Plaques were stained and counted at 8 dpi. The experiment was independently repeated three times, and the data from a single representative experiment are shown. Figure 7 B: HFFs were infected with ganciclovir-resistant HCMV Towne (MOI = 0.1) and treated with the indicated concentrations of MLS8091. Luciferase activity in cell lysates was detected at 72 hpi. The data shown are the mean ± SD of three values from a single experiment. Figure 7 C, 7D: HFFs were infected with HCMV Towne (MOI = 1), and the compound was added or removed at different time points after infection (0, 6, 24, 48, and 72 hours). Luciferase activity in cell lysates was measured at 72 hpi. The experiment was independently repeated twice, and the representative data from a single experiment are shown. Figure 7 E: HFFs were infected with HCMV Towne (MOI = 1), and the expression of viral proteins and cellular β-actin was determined at 72 hpi. The WB data are from a representative experiment. Figure 7 F, 7G: HFFs were infected with HCMV (MOI = 0.1) and treated with MLS8091 (1.5 μM) or ganciclovir (5 μM). Supernatants were collected, and cells were lysed at the indicated time points to isolate DNA. Viral DNA replication in cells ( Figure 7 F) and viral DNA load in supernatants ( Figure 7 G) were determined by real-time PCR. The experiment was independently repeated twice. The data shown are the mean ± SD of three values from a single experiment.

[0074] Figure 8 : NCGC2955 is an early-late inhibitor of HCMV replication. Figure 8 A: HFFs were infected with HCMV TB40 (100 PFU / well) and treated with the indicated concentrations of NCGC2955. Plaques were stained and counted at 8 dpi. The experiment was independently repeated three times, and the data from a single representative experiment are shown. Figure 8 B: HFFs were infected with ganciclovir-resistant HCMV Towne (100 PFU / well) and treated with the indicated concentrations of NCGC2955. Plaques were stained and counted at 8 dpi. The data shown are the mean ± SD of three values from a single experiment. Figure 8C, 8D: HFFs were infected with HCMV Towne (MOI = 1), and compounds were added or removed at different times post - infection (0, 6, 24, 48, and 72 h). Luciferase activity in cell lysates was measured at 72 hpi. The data shown are the mean (mean ± SD) of three independent experiments. Figure 8 E: HFFs were infected with HCMV Towne (MOI = 1), and viral protein expression was determined at 72 hpi. The experiment was independently repeated three times, and data from a single representative experiment are shown. Figure 8 F, 8G: HFFs were infected with HCMV Towne (MOI = 0.1) and treated with 2955 (3 μM) or GCV (5 μM). Supernatants were collected, and cells were lysed at the indicated time points to isolate DNA. Viral DNA replication in cells ([ Figure 8 F) and viral DNA load in supernatants ([ Figure 8 G) were determined by real - time PCR. The data shown are the mean ± SD of four values from two independent experiments.

[0075] Figure 9 : The combination of the newly identified HCMV inhibitor and GCV is additive. ( Figure 9 A - 9E) HFFs were infected with HCMV Towne (MOI = 1) and treated with each compound alone, combinations of each compound at different doses with GCV. For drug combinations, HCMV - infected cells were treated with starting drug concentrations twice the EC 50 value of the single compound and two - fold serial dilutions. Luciferase activity in cell lysates was detected at 72 hpi, and the antiviral activity of the compounds in the drug combinations was calculated by the Bliss model. The solid line represents the observed HCMV inhibition (dose - response), and the dashed line represents the expected HCMV inhibition for each dose of the drug combination. Figure 9 A - 9D: The experiment was independently repeated three times. Data from a single representative experiment are shown. Figure 9 E: The data shown are the mean ± SD of three values from a single experiment.

[0076] Figure 10 : The combination of the newly identified HCMV inhibitor and the HCMV terminase inhibitor Letermovir is additive. ( Figure 10 A - 10E) HFFs were infected with TB40 (100 PFU / well) and treated with each compound alone, combinations of each compound at different doses with Letermovir. For drug combinations, HCMV - infected cells were treated with the EC 50Treat with the starting drug concentration twice the value, followed by treatment with twice serial dilutions. Count the number of plaques in each case at 10 dpi, and calculate the antiviral activity of the compound using the Bliss model. The solid line represents the observed HCMV inhibition (dose response), and the dashed line represents the expected HCMV inhibition for each dose of the drug combination. The data shown are the mean ± SD of three values from a single experiment.

[0077] Figure 11 : Table of Contents Figure: Time of maximum activity of five new inhibitors and GCV during HCMV replication. As described in the "Materials and Methods" section, an addition and removal test was performed for each of the five compounds, and each compound is shown in Figure 5 - 8 C & D. When added and removed, calculate the time when the luciferase activity of each compound is reduced by 75%, and represent it in a unique color. Figure 5 、 6 、7, 8C, and 8D provide separate addition and removal tests. DETAILED DESCRIPTION OF THE INVENTION

[0079] There is a need to improve currently available herpesvirus drugs. For this purpose, a large-scale screening campaign of approximately 400,000 compounds was carried out using a highly sensitive pp28-luciferase HCMV reporter and multiple chemical library collections 20

[0080] Compounds and pharmaceutical compositions for preventing and treating human herpesviruses in subjects in need are disclosed herein.

[0081] The compounds described herein are capable of inhibiting human herpes when administered to a cell or population of cells in an effective amount. In certain embodiments, the cell or population of cells is in a host organism or subject.

[0082] As used herein, the term "human herpesvirus" refers to a DNA virus of the family Herpesviridae known to infect humans. Examples of Herpesviridae species include: (HHV-1) HSV-1 that causes cold sores on the face, HSV-2 (genital herpes) (HHV-2), varicella-zoster virus (HHV-3) that causes chickenpox and shingles, Epstein-Barr virus (HHV4) that causes mononucleosis, HCMV or human cytomegalovirus (CMV or HHV-5), roseolovirus (HHG-6A), herpes lymphotropic virus (HHHV-6B), pityriasis rosea (HHV-7), and Kaposi's sarcoma-associated herpesvirus (HHV-8).

[0083] The present disclosure relates to compounds of Formulas I-V, which can be used to inhibit viral diseases. In some embodiments, the compounds of Formulas I-V can be used to inhibit herpesvirus infections in a subject in need thereof, including mammals such as humans, such as infants, children, and adults, including adults 40 years of age and older.

[0084] The present disclosure also relates to pharmaceutical compositions comprising a compound of Formulas I-V and a pharmaceutical carrier, which can be used to inhibit viral diseases. In some embodiments, the pharmaceutical composition can be used to inhibit herpesvirus infections in mammals (including humans).

[0085] According to one or more embodiments, the present disclosure also relates to pharmaceutical compositions comprising a compound of Formulas I-V, at least one additional bioactive agent, and a pharmaceutical carrier, which can be used to inhibit viral diseases. In some embodiments, the pharmaceutical composition can be used to inhibit herpesvirus infections in mammals (including humans).

[0086] According to one or more embodiments, the present disclosure relates to a method for inhibiting human herpesvirus in a subject, the method comprising: administering to the subject a therapeutically effective amount of a compound of Formulas I-V or a pharmaceutically acceptable composition comprising a compound of Formulas I-V and a pharmaceutically acceptable carrier.

[0087] The present disclosure relates to a method for inhibiting human herpesvirus in a subject in need thereof, the method comprising administering to the subject a compound of Formula I or a pharmaceutical composition thereof or a salt, solvate, or stereoisomer thereof, the pharmaceutical composition comprising a pharmaceutically acceptable carrier:

[0088]

[0089] wherein R 1 is a cycloalkyl, heterocycloalkyl, aryl, or heteroaryl optionally substituted with one or more amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidino, acyl guanidine, having 1 to 10 carbon atoms and linked through a C 1 -C 3 alkyl linkage. In some embodiments, the compound is selected from the group consisting of:

[0090]

[0091]

[0092] The present disclosure relates to a method for inhibiting human herpesvirus in a subject in need thereof, the method comprising administering to the subject a compound of Formula II or a pharmaceutical composition thereof or a salt, solvate, or stereoisomer thereof, the pharmaceutical composition comprising a pharmaceutically acceptable carrier:

[0093]

[0094] Wherein, R 1 is H, and R 2 is independently H or an alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl optionally substituted by one or more alkyl, amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidine, acyl guanidine, having 1 to 10 carbon atoms and linked through C 1 -C 3 alkyl; or R 1 and R 2 together form a C 1 -C 6 heterocycloalkyl or C 1 -C 6 heteroaryl optionally substituted by one or more amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidine, acyl guanidine, having 1 to 10 carbon atoms; and R 3 is H or a cycloalkyl, heterocycloalkyl, aryl or heteroaryl optionally substituted by one or more halogen, CF 3 , CN and NO 2 moieties. In some embodiments, R 2 is a C 1 -C 6 heterocycloalkyl or C 1 -C 6 heteroaryl optionally substituted by one or more alkyl, amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidine, acyl guanidine, having 1 to 10 carbon atoms and linked through C 1 -C 3 alkyl. In some embodiments, the compound is selected from the group consisting of:

[0095]

[0096]

[0097] Disclosed herein is a method for inhibiting human herpesvirus in a subject in need thereof, the method comprising administering to the subject a compound of formula III or a pharmaceutical composition thereof or a salt, solvate or stereoisomer thereof, the pharmaceutical composition comprising a pharmaceutically acceptable carrier:

[0098]

[0099] The present disclosure provides methods for inhibiting human herpesviruses in a subject in need thereof, the methods comprising administering a compound of formula IV or a pharmaceutical composition thereof or a salt, solvate or stereoisomer thereof, the pharmaceutical composition comprising a pharmaceutically acceptable carrier:

[0100]

[0101] The present disclosure provides methods for inhibiting human herpesviruses in a subject in need thereof, the methods comprising administering to the subject a compound of formula V or a pharmaceutical composition thereof or a salt, solvate or stereoisomer thereof, the pharmaceutical composition comprising a pharmaceutically acceptable carrier:

[0102]

[0103] wherein, R 1 is hydrogen, a C1-C6 straight or branched alkyl or a C3-C6 cycloalkyl; and R 2 is an unsubstituted or R 3 - or OR 3 -substituted phenyl, wherein R 3 is hydrogen, a C1-C6 straight or branched alkyl or a C3-C6 cycloalkyl. In some embodiments, R 1 is a C1-C4 straight or branched alkyl; and R 2 is a phenyl substituted with OR 3 wherein R 3 is a C1-C4 straight or branched alkyl. In some embodiments, the compound is

[0104]

[0105] In some embodiments, the methods disclosed herein further comprise administering more than one bioactive agent.

[0106] In some embodiments, the herpesvirus is selected from: human herpesvirus type 1 (HHV-1), human herpesvirus type 2 (HHV2), human herpesvirus type 3 (HHV3), human herpesvirus type 4 (HHV4), human herpesvirus type 5 (CMV or HHV5), human herpesvirus type 6 (HHV6), human herpesvirus type 7 (HHV7), and human herpesvirus type 8 (HHV8). In some embodiments, the herpesvirus is HHV-5.

[0107] The present disclosure provides a pharmaceutical composition comprising a compound of formula I or a salt, solvate or stereoisomer thereof:

[0108]

[0109] wherein, R 1is cycloalkyl, heterocycloalkyl, aryl or heteroaryl optionally substituted by one or more amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidine, acyl guanidine, having 1 to 10 carbon atoms and connected by C 1 -C 3 alkyl linkage. In some embodiments, the compound is selected from the group consisting of:

[0110]

[0111] Disclosed herein is a pharmaceutical composition comprising a compound of Formula II or a salt, solvate or stereoisomer thereof:

[0112]

[0113] wherein, R 1 is H, and R 2 is independently H or alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl optionally substituted by one or more alkyl, amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidine, acyl guanidine, having 1 to 10 carbon atoms and connected by C 1 -C 3 alkyl linkage; or R 1 and R 2 together form a C 1 -C 6 heterocycloalkyl or C 1 -C 6 heteroaryl optionally substituted by one or more amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidine, acyl guanidine, having 1 to 10 carbon atoms; and R 3 is H or cycloalkyl, heterocycloalkyl, aryl or heteroaryl optionally substituted by one or more halogen, CF 3 , CN and NO 2 moieties. In some embodiments, R 2 is a C 1 -C 6 heterocycloalkyl or C 1 -C 6 heteroaryl optionally substituted by one or more amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidine, acyl guanidine, having 1 to 10 carbon atoms and connected by C 1 -C 3 alkyl linkage. In some embodiments, the compound is selected from the group consisting of:

[0114]

[0115]

[0116] The present disclosure relates to a pharmaceutical composition comprising a compound of formula III or a salt, solvate or stereoisomer thereof, and a pharmaceutically acceptable carrier:

[0117]

[0118] The present disclosure relates to a pharmaceutical composition comprising a compound of formula IV or a salt, solvate or stereoisomer thereof, and a pharmaceutically acceptable carrier:

[0119]

[0120] The present disclosure relates to a pharmaceutical composition comprising a compound of formula V or a salt, solvate or stereoisomer thereof, and a pharmaceutically acceptable carrier:

[0121]

[0122] wherein, R 1 is hydrogen, a C1-C6 straight or branched alkyl, or a C3-C6 cycloalkyl; and R 2 is an unsubstituted or R 3 - or OR 3 -substituted phenyl, wherein R 3 is hydrogen, a C1-C6 straight or branched alkyl, or a C3-C6 cycloalkyl. In some embodiments, R 1 is a C1-C4 straight or branched alkyl; and R 2 is a phenyl substituted with OR 3 wherein R 3 is a C1-C4 straight or branched alkyl. In some embodiments, the compound is

[0123]

[0124] The present disclosure relates to a compound of formula I or a salt, solvate or stereoisomer thereof:

[0125]

[0126] wherein, R 1 is a cycloalkyl, heterocycloalkyl, aryl or heteroaryl optionally substituted with one or more amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidino, acyl guanidine, having 1 to 10 carbon atoms and connected by C 1 -C 3 alkyl, provided that the compound cannot be compound A:

[0127]

[0128] In some embodiments of the compound or its salt, solvate or stereoisomer, the compound is selected from:

[0129]

[0130] Disclosed herein are compounds of Formula II or their salts, solvates or stereoisomers:

[0131]

[0132] wherein R 1 is H, and R 2 is independently H or an alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl optionally substituted by one or more alkyl, amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidine, acyl guanidine, having 1 to 10 carbon atoms and linked through C 1 -C 3 alkyl; or R 1 and R 2 together form a C 1 -C 6 heterocycloalkyl or C 1 -C 6 heteroaryl, optionally substituted by one or more amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidine, acyl guanidine, having 1 to 10 carbon atoms; and R 3 is H or a cycloalkyl, heterocycloalkyl, aryl or heteroaryl optionally substituted by one or more halogen, CF 3 , CN and NO 2 moieties, provided that the compound cannot be a compound selected from the group consisting of:

[0133]

[0134] In some embodiments, R 2 is a C 1 -C 6 heterocycloalkyl or C 1 -C 6 heteroaryl, optionally substituted by one or more alkyl, amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidine, acyl guanidine, having 1 to 10 carbon atoms and linked through C 1 -C 3 alkyl.

[0135] Disclosed herein are compounds of Formula V or their salts, solvates or stereoisomers:

[0136]

[0137] Wherein, R 1 is hydrogen, a C1-C6 straight or branched alkyl group, or a C3-C6 cycloalkyl group; and R 2 is an unsubstituted or R 3 or OR 3 substituted phenyl group, wherein R 3 is hydrogen, a C1-C6 straight or branched alkyl group, or a C3-C6 cycloalkyl group. In some embodiments, R 1 is a C1-C4 straight or branched alkyl group; and R 2 is a phenyl group substituted by OR 3 wherein R 3 is a C1-C4 straight or branched alkyl group. In some embodiments, the compound is

[0138]

[0139] The term "aliphatic" is a well-recognized term in the art and includes straight-chain, branched-chain, and cyclic alkanes, alkenes, or alkynes. In certain embodiments, the aliphatic group is straight-chain or branched-chain and has 1 to about 20 carbon atoms.

[0140] The term "alkyl" is well-recognized in the art and includes saturated aliphatic groups, which include straight-chain alkyl groups, branched-chain alkyl groups, cycloalkyl groups (alicyclic groups), alkyl-substituted cycloalkyl groups, and cycloalkyl-substituted alkyl groups. In certain embodiments, the straight-chain or branched-chain alkyl group has about 30 or fewer carbon atoms in its backbone (e.g., for a straight chain it is C 1 -C 30 , and for a branched chain it is C 3 -C 30 ), or about 20 or fewer carbon atoms. Similarly, the cycloalkyl group has about 3 to about 10 carbon atoms in its ring structure, or has about 5, 6, or 7 carbon atoms in the ring structure.

[0141] In addition, the term "alkyl" (or "lower alkyl") includes "unsubstituted alkyl" and "substituted alkyl", the latter referring to an alkyl moiety having substituents that replace hydrogen on one or more carbons of the hydrocarbon backbone. Such substituents can include, for example, halogen, hydroxy, carbonyl (such as carboxy, alkoxycarbonyl, formyl or acyl), thiocarbonyl (such as thioester, thioacetate or thiocarboxylate), alkoxy, phosphoryl, phosphonate, phosphinate, amino, amidino, imine, cyano, nitro, azido, mercapto, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamide, sulfonyl, heterocyclic, aralkyl or aromatic or heteroaromatic moieties. Those skilled in the art will understand that, where appropriate, the substituted portions on the hydrocarbon chain can themselves be substituted. For example, substituents of a substituted alkyl can include substituted and unsubstituted forms of amino, azido, imino, amido, phosphoryl (including phosphonate and phosphinate), sulfonyl (including sulfate, sulfonamide, sulfamoyl and sulfonate) and silyl, as well as ether, alkylthio, carbonyl (including ketone, aldehyde, carboxylate and ester), -CF 3 , -CN, etc. Exemplary substituted alkyls are described below. The cycloalkyl can be further substituted by alkyl, alkenyl, alkoxy, alkylthio, aminoalkyl, carbonyl-substituted alkyl, -CF 3 , -CN, etc.

[0142] The term "aralkyl" is well recognized in the art and includes an aryl group (such as an aromatic or heteroaromatic group).

[0143] The terms "alkenyl" and "alkynyl" are well recognized in the art and refer to moieties containing at least one double bond or triple bond, respectively.

[0144] The term "aryl" is well recognized in the art and includes monocyclic aromatic groups of 5, 6 and 7 members that can contain from 0 to 4 heteroatoms, such as benzene, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine and pyrimidine, etc. The term "heteroatom" generally includes atoms of any element other than carbon or hydrogen. Illustrative heteroatoms include boron, nitrogen, oxygen, phosphorus, sulfur and selenium. An aryl having a heteroatom in the ring structure can also be referred to as "aryl heterocycle" or "heteroaromatic". The aromatic ring can be substituted at one or more ring positions by the above-mentioned substituents, such as halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxy, alkoxy, amino, nitro, mercapto, imino, amido, phosphonate, phosphinate, carbonyl, carboxy, silyl, ether, alkylthio, sulfonyl, sulfonamide, ketone, aldehyde, ester, heterocyclic, aromatic or heteroaromatic moiety, -CF 3, -CN, etc. The term "aryl" also includes polycyclic systems having two or more rings, where two or more carbons are common to two adjacent rings (the rings are "fused rings"), and where at least one ring is an aromatic ring. For example, the other rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, and / or heterocyclic, or rings connected by an acyclic moiety.

[0145] The terms "ortho", "meta", and "para" are well recognized in the art and apply respectively to 1,2-, 1,3-, and 1,4-disubstituted benzene rings. For example, the names 1,2-xylene and ortho-xylene are synonymous.

[0146] The term "heterocyclic group" or "heterocycloalkyl" is well recognized in the art and includes ring structures of about 3 to about 10 members, such as 3-membered rings to about 7-membered rings, having 1 to 4 heteroatoms in the ring structure. The heterocycle can also be polycyclic. Heterocyclic groups include, for example, thiophene, thianthrene, furan, pyran, isobenzofuran, chromene, xanthene, phenoxanthin, pyrroloimidazole, pyrazole, isothiazole, isoxazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine, quinoxaline, isoquinoline, quinoline, phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, pyrimidine, phenanthroline, phenazine, phenopyrazine, phenothiazine, furazan, phenoxazine, pyrrolidine, oxolane, thiolane, oxazole, piperidine, piperazine, morpholine, lactone, lactam (such as azetidinone and pyrrolidone), sultams, sultones, etc. The heterocyclic ring can be substituted at one or more positions with the above substituents, such as halogen, alkyl, aralkyl, alkynyl, cycloalkyl, hydroxy, amino, nitro, mercapto, imino, amido, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, heterocyclic group, aromatic or heteroaromatic moiety, -CD 3 , -CN, etc.

[0147] The terms "polycyclic group" and "polycyclic moiety" are well recognized in the art and include structures having two or more rings (e.g., cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, and / or heterocyclic), where two or more carbons are common to two adjacent rings, e.g., the rings are "fused rings". Rings connected by non-adjacent atoms, e.g., where three or more atoms are common to two rings, are called "bridged" rings. Each ring of the polycycle can be substituted with the above substituents, such as halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxy, amino, nitro, mercapto, imino, amido, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, heterocyclic group, aromatic or heteroaromatic moiety, -CD 3 , -CN, etc.

[0148] As used herein, the term "carbocyclic ring" is well recognized in the art and includes aromatic or non-aromatic rings, wherein each atom of the ring is carbon. The following terms well recognized in the art have the following meanings: "nitro" means -NO 2 ; the term "halogen" means -F, -Cl, -Br or -I; the term "mercapto" means -SH; the term "hydroxyl" or "hydroxyl group" means -OH; the term "sulfonyl" means -SO 2 -.

[0149] The terms "amine" and "amino" are well recognized in the art and include unsubstituted amines and substituted amines. A primary amine bears two hydrogens, a secondary amine bears one hydrogen and another substituent, and both hydrogens of a tertiary amine are substituted. For example, the substituent for one or two hydrogens can be alkyl, alkenyl and aryl, cycloalkyl, cycloalkenyl, heterocycle, polycycle, etc. If both hydrogens are substituted by carbonyl groups, an imide will be formed around the nitrogen.

[0150] The term "alkylamine" includes an amine group as defined above, to which a substituted or unsubstituted alkyl is attached.

[0151] The term "amido" is well recognized in the art as a carbonyl group substituted by an amino group.

[0152] The term "alkylthio" is well recognized in the art and includes an alkyl as defined above, to which a sulfur radical is attached. In certain embodiments, the "alkylthio" moiety is represented as -S-alkyl, -S-alkenyl, -S-alkynyl, etc. Representative alkylthios include methylthio, ethylthio, etc.

[0153] The term "carbonyl" is well recognized in the art and includes the C=O structure. Carbonyls are included in esters; carboxyl groups; formates; thiocarbonyls; thioesters; thiocarboxylic acids; thioformates; ketones; and aldehydes.

[0154] The terms "alkoxy" and "alkoxy group" are well recognized in the art and include an alkyl as defined above, to which an oxygen radical is attached. Representative alkoxys include methoxy, ethoxy, propoxy, tert-butoxy, etc.

[0155] An "ether" is two hydrocarbons covalently linked by oxygen. Thus, the alkyl substituent that makes the alkyl an ether is an alkoxy or alkoxy-like, such as represented by one of -O-alkyl, -O-alkenyl, -O-alkynyl, etc.

[0156] The term "sulfonate" is well recognized in the art and includes a moiety in which the sulfur atom bears two double-bonded oxygens and one single-bonded oxygen.

[0157] The term "sulfate" is well recognized in the art and includes a moiety similar to sulfonate but containing two single-bonded oxygens.

[0158] The terms "sulfonamide", "sulfamoyl", "sulfonyl", and "sulfoxido" are recognized in the art and each encompasses various R group substituents as described herein.

[0159] The terms "phosphoramidite" and "phophonamidite" are recognized in the art.

[0160] The term "selenoalkyl" is recognized in the art and includes an alkyl group having a substituted seleno group attached thereto. Exemplary "selenoethers" that can be substituted on the alkyl group are selected from one of the following: -Se-alkyl, -Se-alkenyl, -Se-alkynyl, etc.

[0161] Substitution can be carried out on the alkenyl and alkynyl groups to produce, for example, aminoalkenyl, aminoalkynyl, amidoalkenyl, iminoalkenyl, iminoalkynyl, thioalkenyl, thioalkynyl, carbonyl-substituted alkenyl or alkynyl.

[0162] Hydrocarbon is a term recognized in the art and includes all permitted compounds having at least one hydrogen atom and one carbon atom. For example, permitted hydrocarbons include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic organic compounds that can be substituted or unsubstituted.

[0163] The term "protecting group" is well known in the art and includes temporary substituents that protect potentially reactive functional groups from undesired chemical transformations. Examples of such protecting groups include esters of carboxylic acids, silyl ethers of alcohols, and acetals and ketals of aldehydes and ketones. The field of protecting group chemistry has been reviewed, for example, by Greene et al., Protective Groups in Organic Synthesis, Second Edition, Wiley, New York, (1991).

[0164] Unless otherwise expressly stated or the context otherwise requires, when an expression (e.g., alkyl, aryl, etc.) appears more than once in any structure, the limitation of each such expression is meant to be independent of the limitations elsewhere in the same structure.

[0165] The terms triflyl, tosyl, mesyl and nonaflyl are well recognized in the art and refer to the trifluoromethanesulfonyl group, the toluenesulfonyl group, the methanesulfonyl group and the nonafluorobutanesulfonyl group, respectively. The terms triflate, tosylate, mesylate and nonaflate are well recognized in the art and refer to the trifluoromethanesulfonate, toluenesulfonate, methanesulfonate and nonafluorobutanesulfonate functional groups and molecules containing said groups, respectively.

[0166] The abbreviations Me, Et, Ph, Tf, Nf, Ts and Ms are well recognized in the art and represent methyl, ethyl, phenyl, triflyl, nonaflyl, tosyl and mesyl, respectively. A more detailed list of abbreviations used by organic chemists of ordinary skill in the art can be found in the first issue of each volume of the Journal of Organic Chemistry; this list generally appears in tabular form under the heading "List of Standard Abbreviations".

[0167] Accordingly, the compounds disclosed herein include the tautomeric forms of the disclosed compounds (including enantiomeric, stereoisomeric and diastereoisomeric forms), as well as their pharmaceutically acceptable salts. The term "pharmaceutically acceptable salts" includes salts commonly used to form alkali metal salts and to form free acid or free base addition salts. Examples of acids that can be used to form pharmaceutically acceptable acid addition salts include inorganic acids (such as hydrochloric acid, sulfuric acid and phosphoric acid) and organic acids (such as maleic acid, succinic acid and citric acid). Other pharmaceutically acceptable salts include salts containing alkali metals or alkaline earth metals (such as sodium, potassium, calcium and magnesium), or salts containing organic bases (such as dicyclohexylamine). For example, suitable pharmaceutically acceptable salts of the disclosed compounds include acid addition salts, which can be formed, for example, by mixing a solution of the disclosed compound with a solution of a pharmaceutically acceptable acid, such as hydrochloric acid, sulfuric acid, methanesulfonic acid, fumaric acid, maleic acid, succinic acid, acetic acid, benzoic acid, oxalic acid, citric acid, tartaric acid, carbonic acid or phosphoric acid. All of these salts can be prepared by conventional methods, for example, by reacting the appropriate acid or base with the corresponding compound disclosed herein.

[0168] Salts formed with free carboxyl groups can also be derived from inorganic bases such as sodium, potassium, ammonium, calcium, or iron hydroxides, and organic bases such as isopropylamine, trimethylamine, 2-ethylaminoethanol, histidine, procaine, etc.

[0169] For use in a drug, the salts of the compounds disclosed herein should be pharmaceutically acceptable salts. However, other salts can also be used to prepare the compounds disclosed herein or their pharmaceutically acceptable salts.

[0170] In addition, the embodiments disclosed herein include hydrates of the compounds disclosed herein. The term "hydrate" includes, but is not limited to, hemihydrate, monohydrate, dihydrate, trihydrate, etc. The hydrates of the compounds disclosed herein can be prepared by contacting the compound with water under suitable conditions to produce the selected hydrate.

[0171] The embodiments disclosed herein also include methods for preparing pharmaceutical products comprising the compounds. The term "pharmaceutical product" refers to a composition (pharmaceutical composition) suitable for medical use as defined herein. A pharmaceutical composition formulated for a specific application and comprising the compounds disclosed herein is regarded as an embodiment thereof.

[0172] The compositions disclosed herein may comprise a carrier. The term "carrier" refers to a diluent, adjuvant, excipient or vehicle with which a therapeutic is administered together.

[0173] With respect to the pharmaceutical compositions described herein, a pharmaceutically acceptable carrier can be any carrier conventionally used and is limited only by physicochemical factors (such as solubility and lack of reactivity with the active compound), as well as by the route of administration. The pharmaceutically acceptable carriers described herein, such as vehicles, adjuvants, excipients and diluents, are well known to those skilled in the art and are readily available to the public. Examples of pharmaceutically acceptable carriers include soluble carriers, such as physiologically acceptable known buffers (such as phosphate buffer), and solid compositions, such as solid carriers or latex beads. Pharmaceutically acceptable carriers are preferably those that are chemically inert to the active agent and those that have no or almost no harmful side effects under the conditions of use.

[0174] The carrier or diluent used herein can be a solid carrier or diluent for solid formulations, a liquid carrier or diluent for liquid formulations, or a mixture thereof.

[0175] Solid carriers or diluents include, but are not limited to: gums, starches (such as corn starch, pregelatinized starch), sugars (such as lactose, mannitol, sucrose, dextrose), cellulosic materials (such as, microcrystalline cellulose), acrylates (such as polymethacrylate), calcium carbonate, magnesium oxide, talc, or mixtures thereof.

[0176] For liquid formulations, pharmaceutically acceptable carriers can be, for example, aqueous or non-aqueous solutions, suspensions, emulsions or oils. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, injectable organic co-solvents, surfactants and injectable organic esters such as ethyl oleate. Aqueous carriers include: for example, water, alcoholic solutions / aqueous solutions, cyclodextrins, emulsions or suspensions, including saline and buffer media.

[0177] Examples of oils are oils of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, olive oil, sunflower seed oil, cod liver oil, sesame oil, cottonseed oil, corn oil, olive, petrolatum and mineral oil. Suitable fatty acids for parenteral preparations include, for example, oleic acid, stearic acid and isostearic acid. Ethyl oleate and isopropyl myristate are examples of suitable fatty acid esters.

[0178] Parenteral carriers (for subcutaneous, intravenous, arterial or intramuscular injection) include, for example, sodium chloride solution, saline solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's solution or fixed oil. Preparations suitable for parenteral administration include, for example, aqueous and non-aqueous solutions, isotonic sterile injection solutions which may contain antioxidants, buffers, bacteriostatic agents and solutes to render the preparation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions containing suspending agents, solubilizers, thickening agents, stabilizers and preservatives.

[0179] Intravenous carriers include, for example, fluids and nutritional supplements, electrolyte supplements such as those based on Ringer's dextrose and the like. Examples are sterile liquids with or without surfactants and other pharmaceutically acceptable adjuvants, such as water and oil. Generally, water, saline, aqueous dextrose solutions and related sugar solutions, and glycols such as propylene glycol or polyethylene glycol may be liquid carriers, especially for injection solutions.

[0180] In addition, in some embodiments, the compounds described herein may further include, for example, binders (e.g., acacia, corn starch, gelatin, carbomer, ethyl cellulose, guar gum, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, polyvinylpyrrolidone); disintegrants (e.g., corn starch, potato starch, alginic acid, silica, croscarmelose sodium, crospovidone, guar gum, sodium starch glycolate); buffers of different pH values and ionic strengths (e.g., Tris-HCl (tris(hydroxymethyl)aminomethane hydrochloride), acetate, phosphate); additives such as albumin or gelatin to prevent adsorption to surfaces; detergents (e.g., Tween 20, Tween 80, Pluronic F68, cholate); protease inhibitors; surfactants (e.g., sodium dodecyl sulfate); penetration enhancers, solubilizers (e.g., cremophor, glycerol, polyethylene glycol, benzlkoniumchloride, benzyl benzoate, cyclodextrin, sorbitan esters, stearic acid); antioxidants (e.g., ascorbic acid, sodium metabisulfite, butylated hydroxyanisole); stabilizers (e.g., hydroxypropyl cellulose, hydroxypropyl methyl cellulose); thickeners (e.g., carbomer, colloidal silica, ethyl cellulose, guar gum); sweeteners (e.g., aspartame, citric acid); preservatives (e.g., thimerosal, benzyl alcohol, parabens); lubricants (e.g., fatty acids, magnesium stearate, polyethylene glycol, sodium dodecyl sulfate); glidants (e.g., colloidal silica); plasticizers (e.g., diethyl phthalate, triethyl citrate); emulsifiers (e.g., carbomer, hydroxypropyl cellulose, sodium dodecyl sulfate); polymer coatings (e.g., poloxamers or poloxamines); coating and film-forming agents (e.g., ethyl cellulose, acrylate, polymethacrylate); and / or adjuvants.

[0181] The choice of carrier will be determined in part by the particular compound and the particular method used to administer the compound. Accordingly, there are a variety of suitable formulations of the pharmaceutical compositions disclosed herein. The following formulations for parenteral, subcutaneous, intravenous, intramuscular, intraarterial, intrathecal, and intraperitoneal administration are exemplary and not limiting. More than one route can be used to administer the compound, and in certain cases, a particular route may provide a more direct and effective response than another route. In some embodiments, in the methods disclosed herein, the compounds or pharmaceutical compositions disclosed herein are administered orally, for example, as tablets, capsules, suspensions, or intravenously.

[0182] Suitable soaps for parenteral preparations may include, for example, fatty alkali metals, ammonium, and triethylamine salts, and suitable detergents may include, for example, (a) cationic detergents such as dimethyldialkylammonium halides and alkylpyridinium halides, (b) anionic detergents such as alkyl, aryl, and olefin sulfonates, alkyl, olefin, ether, and monoglyceride sulfonates, and sulfosuccinates, (c) nonionic detergents such as fatty amine oxides, fatty acid alkanolamides, and polyethylene-polypropylene copolymers, (d) amphoteric detergents such as alkyl-β-aminopropionates and 2-alkylimidazoline quaternary ammonium salts, and (e) mixtures thereof.

[0183] In solution, parenteral preparations generally contain from about 0.5% to about 25% by weight of the compound. Preservatives and buffers may be used. To minimize or eliminate irritation at the injection site, such compositions may contain one or more nonionic surfactants, for example, having a hydrophilic-lipophilic balance value (HLB) of about 12 to about 17. The amount of surfactant in such preparations is generally from about 5% to about 15% by weight. For example, suitable surfactants include polyethylene glycol sorbitan fatty acid esters such as sorbitan monooleate and high molecular weight adducts of ethylene oxide with hydrophobic bases formed by the condensation of propylene oxide and propylene glycol.

[0184] Parenteral preparations may be presented in unit-dose or multi-dose sealed containers, such as ampoules and vials, and may be preserved under lyophilized (freeze-dried) conditions and require only the addition of a sterile liquid excipient for injection (such as water) immediately before use. Immediate injection solutions and suspensions may be prepared from sterile powders, granules, and tablets.

[0185] Injectable preparations are in accordance with the disclosure herein. The requirements for effective pharmaceutical carriers for injectable compositions are well known to those skilled in the art (see Pharmaceutics and Pharmacy Practice, J.B. Lippincott Co., Philadelphia, PA, edited by Banker and Chalmers, pages 238-250 (1982), and ASHP Handbook on Injectable Drugs, Toissel, 4th Edition, pages 622-630 (2009).

[0186] The amount or dose of a compound, salt, solvate, or stereoisomer of any one of the compounds of formulas I, II, III, IV, and V as described above should be sufficient to produce, for example, a therapeutic or prophylactic response in a subject within a reasonable time frame. The dose will be determined by the efficacy of the particular compound and the condition of the person as well as the weight of the person to be treated.

[0187] As described above, the dosage of any of the compounds, salts, solvates or stereoisomers of the compounds of Formulas I, II, III, IV and V will also be determined by the presence, nature and extent of any adverse side effects that may accompany the administration of a particular compound. Generally, the attending physician will consider a variety of factors, such as age, weight, general health, diet, gender, the compound to be administered, the route of administration and the severity of the condition to be treated, to determine the dosage of the compound for treating each individual patient. By way of example and not limitation of the disclosure herein, the dosage of the compound may be from about 0.001 to about 1000 mg / kg of the weight of the subject to be treated per day.

[0188] In some embodiments, the term "administer" means introducing a compound disclosed herein into a subject, e.g., a subject undergoing treatment for a disease, and allowing the compound to contact one or more disease-related cells or cell populations in the body. In some embodiments, the host cell or host cell population in the host can be any cell or cell population that can selectively bind by binding to an antigen of the compounds of Formulas I, II, III, IV and V described above. Those skilled in the art will understand that the host cell can be a cell infected with a virus.

[0189] According to some embodiments, the disclosure herein provides a method for inhibiting human herpesvirus in a subject, the method comprising: administering to the subject a therapeutically effective amount of a pharmaceutically acceptable composition comprising one or more of the above compounds and a pharmaceutically acceptable carrier.

[0190] According to some embodiments, the disclosure herein provides a method for treating a medical condition in a subject (including but not limited to inhibiting human herpesvirus), the method comprising: administering to the subject a therapeutically effective amount of a pharmaceutically acceptable composition comprising one or more of the above compounds, a pharmaceutically acceptable carrier and at least one other bioactive agent.

[0191] According to some embodiments, the disclosure herein provides a method for inhibiting or treating human herpesvirus in a subject, the method comprising: administering to the subject a therapeutically effective amount of a pharmaceutically acceptable composition comprising one or more of the above compounds and a pharmaceutically acceptable carrier.

[0192] According to some embodiments, the disclosure herein provides a method for inhibiting or treating human herpesvirus (e.g., inhibiting human herpesvirus replication) in a subject, the method comprising: administering to the subject a therapeutically effective amount of a pharmaceutically acceptable composition comprising one or more of the above compounds, at least one bioactive agent and a pharmaceutically acceptable carrier.

[0193] "Inhibiting herpes virus" or "inhibition of herpes virus" means reducing or preventing viral replication, treating symptoms caused by viral infection (such as hepatitis or pneumonia), reducing the viral load in blood or plasma, and / or reducing persistent viremia.

[0194] "Administering treatment" or "treatment" is a term recognized in the art and includes curing as well as alleviating at least one symptom of any condition or disease. Treatment includes reducing the likelihood of an animal susceptible to a disease, disorder or condition, but not yet diagnosed with the disease, disorder or condition, from developing the disease, disorder or condition; inhibiting a disease, disorder or condition, such as impeding its progression; and alleviating a disease, disorder or condition, such as causing regression of the disease to any degree; inhibiting a disease, disorder or condition, such as impeding its progression; and alleviating a disease, disorder or condition even if the underlying pathophysiology is not affected or other symptoms remain at the same level.

[0195] In some embodiments, the pharmaceutical compositions disclosed herein can be used to treat a viral infection in a subject. In some embodiments, the viral infection is a herpes virus infection, e.g., a CMV infection.

[0196] In some embodiments, the pharmaceutical compositions disclosed herein can be combined with one or more additional bioactive agents, including, for example, antiviral agents.

[0197] "Prophylactic / preventive" or "therapeutic / treatment" treatment is recognized in the art and includes administering to a host one or more of the subject compositions. If administered before the clinical manifestation of an unwanted condition (e.g., a disease or other unwanted state in a host animal), the treatment is prophylactic, i.e., it protects the host against the development of the unwanted condition, whereas if administered after the manifestation of the unwanted condition, the treatment is therapeutic (i.e., its purpose is to reduce, alleviate or stabilize the existing unwanted condition or its side effects).

[0198] Bioactive agents can vary widely depending on the intended use of the composition. The term "active" is well recognized in the art and refers to any moiety of a biological, physiological, or pharmaceutical active substance that acts locally or systemically in an object. Examples of bioactive agents that may be referred to as "drugs" are described in well-known literature such as the Merck Index, Physicians' Desk Reference, and Pharmacological Basis of Therapeutics, and include, but are not limited to: medicaments; vitamins; mineral supplements; substances for treating, preventing, diagnosing, curing, or alleviating a disease or illness; substances that affect the structure or function of the body; or prodrugs that become biologically active or more active when placed in a physiological environment. Bioactive agents can be used in various forms and are capable of causing the subject composition to be released into, for example, adjacent tissues or fluids after administration to an object. In some embodiments, bioactive agents can be used in crosslinked polymer matrices, for example, to promote chondrogenesis. In other embodiments, bioactive agents can be used in crosslinked polymer matrices to treat, alleviate, inhibit, or prevent a disease or condition, for example, in conjunction with promoting chondrogenesis.

[0199] Further examples of bioactive agents include, but are not limited to: enzymes, receptor antagonists or agonists, hormones, growth factors, autologous bone marrow, antibiotics, antimicrobials, and antibodies. The term "bioactive agent" is also intended to encompass various cell types and genes that can be incorporated into the compounds and compositions disclosed herein.

[0200] In certain embodiments, the subject composition can comprise from about 1 wt% to about 75 wt% or more of the total composition, or can contain about 2.5%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, or 70% of the bioactive agent.

[0201] Non-limiting examples of bioactive agents include the following: adrenergic blocking agents, anabolic agents, androgenic steroids, antacids, anti-asthmatic agents, antiallergenic materials, anti-cholesterolemic and anti-lipid agents, anti-cholinergics and sympathomimetics, anti-coagulants, anti-convulsants, anti-diarrheal, anti-emetics, antihypertensive agents, anti-infective agents, anti-inflammatory agents such as steroids, nonsteroidal anti-inflammatory agents, anti-malarials, anti-manic agents, anti-nauseants, antineoplastic agents, anti-obesity agents, anti-parkinsonian agents, anti-pyretic and analgesic agents, anti-spasmodic agents, anti-thrombotic agents, anti-uricemic agents, anti-anginal agents, antihistamines, anti-tussives, appetite suppressants, benzophenanthridine alkaloids, biologicals, cardioactive agents, cerebralDilators, coronary dilators, decongestants, diuretics, diagnostic agents, erythropoietic agents, estrogens, expectorants, gastrointestinal sedatives, agents, hyperglycemic agents, hypnotics, hypoglycemic agents, ion exchange resins, laxatives, mineral supplements, mitotics, mucolytic agents, growth factors, neuromuscular drugs, nutritional substances, peripheral vasodilators, progestational agents, prostaglandins, psychic energizers, psychotropics, sedatives, stimulants, thyroid and anti-thyroid agents, tranquilizers, uterine relaxants, vitamins, antigenic materials and prodrugs.

[0202] Specific examples of bioactive agents available in the above categories include: (a) anti-tumor drugs, such as androgen inhibitors, antimetabolites, cytotoxic agents, and immunomodulators; (b) cough suppressants, such as dextromethorphan, hydrobromide, noscapine, carbetapentane citrate, and chlophedianol hydrochloride; (c) antihistamines, such as chlorpheniramine phenindamine tartrate, pyrilamine doxylamine succinate, and phenyltoloxamine citrate; (d) decongestants, such as hydrochloride, phenylpropanolamine hydrochloride, pseudoephedrine hydrochloride, and ephedrine; (e) various alkaloids, such as codeine phosphate, codeine sulfate, and morphine; (f) mineral supplements, such as potassium chloride, zinc chloride, calcium carbonate, magnesium oxide, and other alkali metal and alkaline earth metal salts; (g) ion exchange resins, such as N-acetylprocainamide; (i) antipyretics and analgesics, such as acetaminophen, aspirin, and ibuprofen; appetite suppressants, such as phenyl-propanol amine or caffeine; (k) expectorants, such as guaifenesin; (l) antacids, such as aluminum hydroxide and magnesium hydroxide; biological products, such as peptides, polypeptides, proteins, and amino acids, hormones, interferons, or cytokines, and other bioactive peptide compounds, such as calcitonin, ANF, EPO, and insulin;(n) Anti-infective agents, such as antifungal agents, antiviral agents, preservatives and antibiotics; and (m) desensitizing agents and anti-allergen materials, for example materials for vaccine applications.;

[0203] More specifically, non-limiting examples of available bioactive agents include the following therapeutic categories: analgesics, such as nonsteroidal anti-inflammatory drugs, opiate agonists, and salicylates; antihistamines, such as H1-blockers and H2-blockers; anti-infective agents, such as antihelmintics, antianaerobics, antibiotics, aminoglycoside antibiotics, antifungal antibiotics, cephalosporin antibiotics, macrolide antibiotics, miscellaneous antibiotics, penicillin antibiotics, quinolone antibiotics, sulfonamide antibiotics, tetracycline antibiotics, antimycobacterials, antituberculosis antimycobacterials, antiprotozoals, antimalarial antiprotozoals, antiviral agents, anti-retroviral agents, scabicides, and urinary anti-infectives; anti-tumor agents, such as alkylating agents, nitrogen mustard alkylating agents, nitrosourea alkylating agents, antimetabolites, purine analog antimetabolites;Pyrimidine analog antimetabolites; hormonal antineoplastics, natural antineoplastics, antibiotic natural antineoplastics, and vinca alkaloid natural antineoplastics; autonomic agents, such as anticholinergics, antimuscarinic anticholinergics, ergot alkaloids, parasympathomimetics, cholinergic agonist parasympathomimetics, cholinesterase inhibitor parasympathomimetics, sympatholytics, α-blocker sympatholytics, sympatholytics, sympathomimetics, and adrenergic agonist sympathomimetics;Cardiovascular agents, such as antianginals, calcium channel blocker antianginals, nitrate antianginals, antiarrhythmics, cardiac glycoside antiarrhythmics, class I antiarrhythmics, class II antiarrhythmics, class III antiarrhythmics, class IV antiarrhythmics, antihypertensive agents, α-blocker antihypertensives, angiotensin-converting enzyme inhibitor (ACE inhibitor) antihypertensives, β-blocker antihypertensives, calcium-channel blocker antihypertensives, central-acting adrenergic antihypertensives, diuretic antihypertensive agents, peripheral vasodilator antihypertensives, antilipemics, bile acid sequestrant antilipemics, reductase inhibitor antilipemics, inotropes, cardiac glycoside inotropes, and thrombolytic agents;Dermatological agents, such as antihistamines, anti-inflammatory agents, corticosteroid anti-inflammatory agents, anesthetics, topical anti-infectives, topical anti-infectives, antiviral topical anti-infectives, and topical antineoplastics; electrolytic and renal agents, such as acidifying agents, alkalinizing agents, diuretics, carbonic anhydrase inhibitor diuretics, loop diuretics, osmotic diuretics, potassium-sparing diuretics, thiazide diuretics, electrolyte replacements, and uricosuric agents; enzymes, such as pancreatic enzymes and thrombolytic enzymes;Gastrointestinal agents, such as antidiarrheals, antiemetics, gastrointestinal anti-inflammatory agents, salicylate gastrointestinal anti-inflammatory agents, antacid anti-ulcer agents, gastric acid-pump inhibitor anti-ulcer agents, gastric mucosal anti-ulcer agents, H2-blocker anti-ulcer agents, cholelitholytic agents, digestants, emetics, laxatives and stool softeners, as well as prokinetic agents; general anesthetics, such as inhalation anesthetics, halogenated inhalation anesthetics, intravenous anesthetics, barbiturate intravenous anesthetics, benzodiazepine intravenous anesthetics and opiate agonist intravenous anesthetics;Hematological agents, such as antianemia agents, hematopoietic antianemia agents, coagulation agents, anticoagulants, hemostatic coagulation agents, platelet inhibitor coagulation agents, thrombolytic enzyme coagulation agents, and plasma volume expanders; hormones and hormone modulators, such as abortifacients, adrenal agents, corticosteroid adrenal agents, androgens, anti-androgens, antidiabetic agents, sulfonylurea antidiabetic agents, antihypoglycemic agents, oral contraceptives, progestin contraceptives, estrogens, fertility agents, oxytocics, parathyroid agents, pituitary hormones, progestins, antithyroid agents, thyroid hormones, and tocolytics; immunobiological agents, such as immunoglobulins, immunosuppressives, toxoids, and vaccines; local anesthetics, for example, amide local anesthetics and ester local anesthetics;Musculoskeletal agents, such as anti-gout anti-inflammatory agents, corticosteroid anti-inflammatory agents, gold compound anti-inflammatory agents, immunosuppressive anti-inflammatory agents, nonsteroidal anti-inflammatory drugs, salicylate anti-inflammatory agents, skeletal muscle relaxants, neuromuscular blocker skeletal muscle relaxants, and reverse neuromuscular blocker skeletal muscle relaxants; neurological agents, such as anticonvulsants, barbiturate anticonvulsants, benzodiazepine anticonvulsants, anti-migraine agents, anti-parkinsonian agents, anti-vertigo agents, opiate agonists, and opiate antagonists;Ophthalmic agents, such as anti-glaucoma agents, anti-glaucoma agents, mitotics, anti-glaucoma agents, mydriatics, adrenergic agonist mydriatics, antimuscarinic mydriatics, ophthalmic anesthetics, ophthalmic anti-infectives, ophthalmic aminoglycoside anti-infectives, ophthalmic macrolide anti-infectives, ophthalmic quinolone anti-infectives, ophthalmic sulfonamide anti-infectives, ophthalmic tetracycline anti-infectives, ophthalmic anti-inflammatory agents, ophthalmic corticosteroid anti-inflammatory agents, and ophthalmic nonsteroidal anti-inflammatory drugs;Psychotropic agents, such as antidepressants, heterocyclic antidepressants, monoamine oxidase inhibitors, selective serotonin re-uptake inhibitors, tricyclic antidepressants, antimanics, antipsychotics, phenothiazine antipsychotics, anxiolytics, sedatives and hypnotics, barbiturate sedatives and hypnotics, benzodiazepine anxiolytics, sedatives and hypnotics, and psychostimulants; respiratory agents, such as antitussives, bronchodilators, adrenergic agonist bronchodilators, antimuscarinic bronchodilators, expectorants, mucolytic agents, respiratory anti-inflammatory agents and respiratory corticosteroid anti-inflammatory agents; toxicology agents, such as antidotes, heavy agents, substance abuse agents, deterrent substance abuse agents and withdrawal substance abuse agents; minerals; and vitamins, for example, vitamin A, vitamin B, vitamin C, vitamin D, vitamin E, and K.;

[0204] Other classes of bioactive agents from the above classes include: (1) general analgesics such as lidocaine, other "caine" analgesics or their derivatives, and non-steroidal anti-inflammatory drug (NSAID) analgesics including diclofenac, ibuprofen, ketoprofen, and naproxen; (2) opiate agonist analgesics such as codeine, fentanyl, hydromorphone, and morphine; (3) salicylate analgesics such as aspirin (ASA) (enteric-coated ASA); (4) H1-blocker antihistamines such as clemastine and terfenadine; (5) H2-blocker antihistamines such as cimetidine, famotidine, nizadine, and ranitidine; (6) anti-infectives such as mupirocin; (7) anti-anaerobic anti-infectives such as chloramphenicol and clindamycin; (8) anti-fungal antibiotic anti-infectives such as amphotericin b, clotrimazole, fluconazole, and ketoconazole; (9) macrolide antibiotic anti-infectives such as azithromycin and erythromycin; (10) miscellaneous antibiotic anti-infectives such as imipenem; penicillin, (11) antibiotic anti-infectives such as nafcillin, oxacillin, penicillin G, and penicillin V;(12) Quinolone antibiotic anti-infectives, such as ciprofloxacin and norfloxacin; (13) Tetracycline antibiotic anti-infectives, such as doxycycline, minocycline and tetracycline; (14) Antituberculosis antimycobacterial anti-infectives, such as isoniazid and rifampin; (15) Antiprotozoal anti-infectives, such as atovaquone and dapsone; (16) Antimalarial anti-protozoal anti-infectives, such as chloroquine and pyrimethamine; (17) Anti-retroviral anti-infectives, such as ritonavir and zidovudine; (18) Anti-viral anti-infectives, such as acyclovir, ganciclovir, interferon-y and rimantadine; (19) Alkylating antineoplastic agents, such as carboplatin and cisplatin; (20) Nitrosourea alkylating antineoplastic agents, such as carmustine (BCNU); (21) Antimetabolite antineoplastic agents, such as methotrexate; (22) Pyrimidine analog antineoplastic agents, such as fluorouracil (S-FU) and gemcitabine;(23) Hormonal antineoplastics, such as goserelin, leuprolide, and tamoxifen; (24) Natural antineoplastics, for example, aldesleukin, interleukin-2, docetaxel, etoposide, interferon; paclitaxel, other taxane derivatives, and tretinoin (ATRA); (25) Antibiotic natural antineoplastics, such as bleomycin, dactinomycin, daunorubicin, doxorubicin, and mitomycin; (26) Vinca alkaloid natural antineoplastics, such as vinblastine and vincristine; (27) Autonomic agents, for example, nicotine; (28) Anticholinergic autonomic agents, such as benztropine and trihexyphenidyl; (29) Anticholinergic autonomic agents, such as atropine and oxybutynin; (30) Ergot alkaloid autonomic agents, for example, bromocriptine; (31) Cholinergic agonist parasympathomimetics, for example, pilocarpine; (32) Cholinesterase inhibitor parasympathomimetics, for example, pyridostigmine;(33) α-blocker sympatholytics, for example, prazosin; (34) D-blocker sympatholytics, for example, atenolol; (35) adrenergic sympathomimetics, such as albuterol and dobutamine; (36) cardiovascular agents, such as aspirin (ASA) (enteric-coated ASA); (37) D-blocker antianginals, for example, atenolol and propranolol; (38) calcium channel blocker antianginals, for example, nifedipine and verapamil; (39) nitrate antianginals, for example, isosorbide dinitrate (ISDN); (40) cardiac glycoside antiarrhythmics, for example, (41) class I antiarrhythmics, such as lidocaine, mexiletine, phenytoin, procainamide and quinidine; (42) class II antiarrhythmics, such as atenolol, metoprolol, propranolol and timolol; (43) class III antiarrhythmics, such as amiodarone; (44) class IV antiarrhythmics, such as diltiazem and verapamil; (45) antihypertensives, for example, prazosin;(46) Angiotensin-converting enzyme inhibitor (ACE inhibitor) antihypertensives, for example, captopril and enalapril; (47) Antihypertensives, for example, atenolol, metoprolol, nadolol, and propranolol; (48) Calcium-channel blocker antihypertensive agents, for example, diltiazem and nifedipine; (49) Central-acting adrenergic antihypertensives, for example, clonidine and methyldopa; (50) Diuretic antihypertensive agents, for example, amiloride, furosemide, hydrochlorothiazide (HCTZ), and spironolactone; (51) Peripheral vasodilator antihypertensives, for example, minoxidil; (52) Antilipemics, for example, gemfibrozil and probucol; (53) Bile acid sequestrant antilipemics, for example, cholestyramine; (54) Reductase inhibitor antilipemics, for example, lovastatin and pravastatin; (55) Inotropes, for example, amrinone, dobutamine, and dopamine;(56) Cardiac glycoside inotropes, for example, (57) thrombolytic agents, for example, alteplase, anistreplase, streptokinase, and urokinase; (58) dermatological agents, such as colchicine, isotretinoin, methotrexate, minoxidil, tretinoin, (59) dermatological corticosteroid anti-inflammatory agents, such as betamethasone and dexamethasone; (60) antifungal topical anti-infectives, such as amphotericin, clotrimazole, miconazole, and nystatin; (61) antiviral topical anti-infectives, such as acyclovir; (62) topical antineoplastics, for example, (63) electrolytic and renal agents, such as lactulose; (64) loop diuretics, for example, furosemide; (65) potassium-sparing diuretics, for example, triamterene; (66) thiazide diuretics, for example, hydrochlorothiazide (HCTZ); (67) uricosuric agents, for example, probenecid; (68) enzymes and (69) thrombolytic enzymes, for example, alteplase, anistreplase, streptokinase, and urokinase;(70) Antiemetics, for example, prochlorperazine; (71) Salicylate gastrointestinal anti-inflammatory agents, for example, sulfasalazine; (72) Gastric acid-pump inhibitor anti-ulcer agents, for example, omeprazole; (73) H2-blocker anti-ulcer agents, such as cimetidine, famotidine, nizadine, and ranitidine; (74) Digestants, for example, pancrelipase; (75) Prokinetic agents, for example, erythromycin; (76) Opiate agonist intravenous anesthetics, for example, fentanyl; (77) Hematopoietic antianemia agents, such as (G-CSF) and (GM-CSF); (78) Coagulation agents, for example, Factor 1-10 (AlIF 1-10); (79) Anticoagulants, for example, warfarin; (80) Thrombolytic enzyme coagulation agents, for example, alteplase, anistreplase, streptokinase, and urokinase; (81) Hormones and hormone regulators, for example, bromocriptine; (82) Abortifacients, for example, methotrexate; (83) Antidiabetic agents, for example, insulin; (84) Oral contraceptives, for example, estrogen and progestin;(85) Progestin contraceptives, for example, levonorgestrel and norgestrel; (86) Estrogens, for example, conjugated estrogens, diethylstilbestrol (DES), estrogens (estradiol, estrone, and estropipate sulfate); (87) Fertility agents, for example, clomiphene, human chorionic gonadotropin (HCG), and menotropins; (88) Parathyroid agents, for example, calcitonin; (89) Pituitary hormones, for example, desmopressin, goserelin, oxytocin, and vasopressin (ADH); (90) Progestins, for example, medroxyprogesterone, norethindrone, and progesterone; (91) Thyroid hormones, for example, levothyroxine; (92) Immunobiologic agents, for example, interferon β-1b and interferon γ-1b; (93) Immunoglobulins, for example, immunoglobulin 1M, IMIG, IGIM, and immunoglobulin IVIG; (94) Amide local anesthetics, for example, lidocaine; (95) Ester local anesthetics, for example, benzocaine and procaine;(96) Musculoskeletal corticosteroid anti-inflammatory agents, such as beclomethasone, betamethasone, cortisone, dexamethasone, hydrocortisone, and prednisone; (97) Musculoskeletal anti-inflammatory immunosuppressives, such as azathioprine, cyclophosphamide, and methotrexate; (98) Musculoskeletal nonsteroidal anti-inflammatory drugs, such as diclofenac, ibuprofen, ketoprofen, ketorlac, and naproxen; (99) Skeletal muscle relaxants, for example, diazepam; (100) Reverse neuromuscular blocker skeletal muscle relaxants, for example, pyridostigmine; (101) Neurological agents, such as nimodipine, riluzole, tacrine, and ticlopidine; (102) Anticonvulsants, such as carbamazepine, gabapentin, lamotrigine, phenytoin, and valproic acid; (103) Barbiturate anticonvulsants, such as phenobarbital and primidone;(104) Benzodiazepine anticonvulsants, such as clonazepam, diazepam, and lorazepam; (105) Anti-migraine agents, such as bromocriptine, levodopa, carbidopa, and pergolide; (106) Anti-vertigo agents, such as meclizine; (107) Opiate agonists, such as codeine, fentanyl, hydromorphone, methadone, and morphine; (108) Opiate antagonists, such as naloxone; (109) Anti-glaucoma agents, such as timolol; (110) Mitotic anti-glaucoma agents, such as pilocarpine; (111) Ophthalmic aminoglycoside anti-infectives, such as gentamicin, neomycin, and tobramycin; (112) Ophthalmic quinolone anti-infectives, such as ciprofloxacin, norfloxacin, and ofloxacin; (113) Ophthalmic corticosteroid anti-inflammatory agents, such as dexamethasone and prednisolone; (114) Ophthalmic nonsteroidal anti-inflammatory drugs, such as diclofenac; (115) Anti-psychotics, such as clozapine, haloperidol, and risperidone;(116) Benzodiazepine anxiolytics, sedatives and hypnotics, such as clonazepam, diazepam, lorazepam, oxazepam and prazepam; (117) Psychostimulants, such as methylphenidate and pemoline; (118) Such as codeine; (119) Bronchodilators, such as, (120) Adrenergic agonist bronchodilators, such as albuterol; (121) Respiratory corticosteroid anti-inflammatory agents, such as dexamethasone; (122) Antidotes, such as flumazenil and naloxone; (123) Heavy metal agents, such as penicillamine; (124) Deterrent substance abuse agents, such as disulfiram, naltrexone and nicotine; (125) Withdrawal substance abuse agents, such as bromocriptine; (126) Minerals, such as iron, calcium and magnesium; (127) Vitamin B compounds, such as cyanocobalamin (vitamin B12) and niacin (vitamin B3); (128) Vitamin C compounds, such as ascorbic acid; and (129) Vitamin D, such as calcitriol.

[0205] In addition, recombinant or cell-derived proteins can be used, such as recombinant β-glucan; bovine immunoglobulin concentrate; bovine superoxide dismutase; a preparation containing fluorouracil, adrenaline, and bovine collagen; recombinant hirudin (r-Hir), HIV-1 immunogen; recombinant human growth hormone recombinant EPO (r-EPO); gene-activated EPO (GA-EPO); recombinant human hemoglobin (r-Hb); recombinant human mecasermin (r-1GF-1); recombinant interferon α; lenograstim (G-CSF); olanzapine; recombinant thyroid-stimulating hormone (r-TSH); and topotecan.

[0206] In addition, peptides, proteins, and other macromolecules listed below can be used, such as interleukins 1 to 18, including mutants and analogs; interferons a, y, which can be used for cartilage regeneration, hormone releasing hormone (LHRH) and analogs, gonadotropin releasing hormone transforming growth factor (TGF); fibroblast growth factor (FGF); tumor necrosis factor-α; nerve growth factor (NGF); growth hormone releasing factor (GHRF), epidermal growth factor (EGF), connective tissue activating osteogenic factor, fibroblast growth factor homologous factor (FGFHF); hepatocyte growth factor (HGF); insulin growth factor (IGF); invasion inhibitory factor-2 (IIF-2); bone morphogenetic proteins 1-7 (BMP 1-7); somatostatin; thymosin-a-y-globulin; superoxide dismutase (SOD); and complement factors, as well as bioactive analogs, fragments, and derivatives of these factors (such as growth factors).

[0207] Members of the transforming growth factor (TGF) supergene family are multifunctional regulatory proteins that can be incorporated into a polymeric matrix. Members of the TGF supergene family include beta-transforming growth factor (e.g., TGF-β1, TGF-β2, TGF-β3); bone morphogenetic proteins (e.g., BMP-1, BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8, BMP-9); heparin-binding growth factors (e.g., fibroblast growth factor (FGF), epidermal growth factor (EGF), platelet-derived growth factor (PDGF), insulin-like growth factor (IGF)) (e.g., inhibin A, inhibin B), growth differentiation factors (e.g., GDF-1); and activins (e.g., activin A, activin B, activin AB). Growth factors can be isolated from natural or native sources, such as from mammalian cells, or can be prepared by synthetic methods, such as by recombinant DNA technology or various chemical processes. In addition, analogs, fragments, or derivatives of these factors can be used, provided that they exhibit at least some of the biological activities of the native molecule. For example, analogs can be prepared by expressing genes altered by site-specific mutagenesis or other genetic engineering techniques.

[0208] The methods, pharmaceutical compositions, and compounds disclosed herein can be combined to provide one, two, or more of the compounds disclosed herein, optionally in combination with, for example, maribavir, foscarnet, GCV, and / or letermovir.

[0209] Various forms of bioactive agents can be used. These bioactive agents include, but are not limited to: uncharged molecules, molecular complexes, salts, ethers, esters, amides, prodrug forms, etc., which are bioactivated when implanted, injected, or otherwise placed in a subject.

[0210] The following examples are included to guide a representative implementation of the disclosed subject matter for those of ordinary skill in the art. Based on the present disclosure and the general level of skill in the art, those skilled in the art will understand that the following examples are for illustration only and that various changes, modifications, and variations can be made without departing from the scope of the present disclosure. The following synthetic descriptions and specific examples are for illustrative purposes only and should not be construed as limiting the preparation of the compounds of the present disclosure by any other method. Examples

[0211] Example 1

[0212] Materials and Methods

[0213] Compounds: These compounds were identified by high-throughput screening (HTS) and then verified using multiple HCMV strains, multiple antiviral assays, and novel compound preparations. In addition to the compounds obtained from the library used in the screening, NFU1827 was synthesized in the Department of Biochemistry, Program in Chemical Biology at the Medical College of Wisconsin, MLS8091 was purchased from Princeton Biomolecular Research, Inc., MLS8969 was purchased from ChemBridge Corporation in San Diego, California, MLS8554 was purchased from InterBioScreen STOCK1N-59867, and NCGC2955 was synthesized by the Chemistry Core, Johns Hopkins University School of Medicine. Using a Phenomenex Luna C18 3.0 x 75 mm column, a 7-minute gradient of 4 - 100% ACN in H 2 O containing 0.05% v / v TFA, or using a Higgins Analytical, Inc. Targa C18 5 μm 4.6 x 150 mm column, a 30-minute gradient of 0 - 100% ACN in H 2 O containing 0.01% TFA, and absorbance detection or evaporative light scattering detection at 254 nm, the purity of all synthetic and purchased compounds used in biological assays was determined by HPLC. The NMR and MS data of all compounds used in biological assays were consistent with their structures, and the purity determined by HPLC was > 95%.

[0214] The compounds were dissolved in dimethyl sulfoxide (DMSO), and 50 and 10 mM stock solutions were stored at -80 °C. Ganciclovir was purchased from Sigma Aldrich (St. Louis, Missouri), and a 10 mM stock solution was prepared in ddH 2 O.

[0215] Synthesis of NCGC2955: tert-Butyl 4-(isopropylcarbamoyl)piperidine-1-carboxylate (1). To a solution of 1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid 35 (3.94 g, 17.2 mmol, 1.0 equiv) in CH 2 Cl2 (39 mL) of solution was added with N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (3.46 g, 18.0 mmol, 1.05 equiv), 1-hydroxybenzotriazole monohydrate (2.83 g, 18.5 mmol, 1.075 equiv) and isopropylamine (5.1 mL, 60.15 mmol, 3.5 equiv). After stirring at room temperature for 21 h, the reaction was diluted with CH 2 Cl 2 and washed with 5% aqueous HCl solution. The aqueous layer was extracted with CH 2 Cl 2 (3 x 20 mL). The organic layers were combined, dried over anhydrous MgSO 4 and concentrated in vacuo. Purification by flash chromatography (0 to 100% EtOAc / hexane) afforded 2.95 g of the solid compound 1 in 63% yield. 1 1H NMR (500 MHz, CDCl 3 ) δ 5.28 (d, J = 6.45 Hz, 1H), 4.01 - 4.25 (m, 3H), 2.73 (br.s., 2H), 2.16 (tt, J = 3.69, 11.63 Hz, 1H), 1.79 (d, J = 11.32 Hz, 2H), 1.61 (dq, J = 4.24, 12.42 Hz, 2H), 1.45 (s, 9H), 1.14 (d, J = 6.60 Hz, 6H).

[0216] N-Isopropylpiperidine-4-carboxamide trifluoroacetate (2). At room temperature, to a solution of compound 1 (2.95 g, 10.9 mmol) in CH 2 Cl 2 (5 mL) was added triisopropylsilane (250 μL) and trifluoroacetic acid (3 mL). After stirring at room temperature for 24 h, the volatiles were removed in vacuo to afford a viscous slurry, which could be used without further purification. 1H NMR (500 MHz, CDCl3) δ 4.06 (qd, J = 6.86, 13.38 Hz, 1H), 3.57 (d, J = 12.89 Hz, 2H), 3.10 (br.s., 2H), 2.45 - 2.63 (m, 1H), 2.05 - 2.16 (m, 4H), 1.18 (d, J = 6.60 Hz, 6H).

[0217] Methyl 4-(4-chlorobenzyl)-4H-thieno[3,2-b]pyrrole-5-carboxylate (3). At room temperature, to methyl 4H-thieno[3,2-b]pyrrole-5-carboxylate in anhydrous DMF (10 mL) 36(1.44 g, 7.99 mmol, 1.0 equiv) and cesium carbonate (3.91 g, 12.0 mmol, 1.5 equiv) was added in one portion to a suspension of 4-chlorobenzyl bromide (1.97 g, 9.59 mmol, 1.2 equiv). After stirring at room temperature until completion (ca. 4 h), the reaction was diluted with water and extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine, dried over anhydrous MgSO 4 and concentrated in vacuo. Purification by flash chromatography (3 m 0%, 6 m gradient 0 to 50%, 2 m 50% EtOAc / hexanes) afforded 2.40 g of the methyl ester 3 as a pale yellow solid in 98% yield. 1H NMR (500 MHz, CDCl3) δ 7.33 (d, J = 5.34 Hz, 1H), 7.21 - 7.29 (m, 3H), 7.05 (d, J = 8.17 Hz, 2H), 6.85 (d, J = 5.50 Hz, 1H), 5.71 (s, 2H), 3.83 (s, 3H).

[0218] 4-(4-Chlorobenzyl)-4H-thieno[3,2-b]pyrrole-5-carboxylic acid (4). A solution of lithium hydroxide monohydrate in water (12 mL) was added in one portion to a mixture of the methyl ester 3 (3.58 g, 11.7 mmol, 1.0 equiv) in THF (1252 mL) and MeOH (12 mL). After rapid magnetic stirring at room temperature for 2 days, the starting material was consumed and the organic solvents were removed in vacuo. The remaining aqueous solution was diluted with water (ca. 20 mL) and the pH was adjusted to pH = 1 with aqueous concentrated HCl with vigorous magnetic stirring. A precipitate formed during acidification, was collected by vacuum filtration, washed with water and dried in vacuo to afford 3.34 g of compound 4 in 98% yield. 1H NMR (500 MHz, DMSO-d6) δ 12.57 (br.s., 1H), 7.56 (d, J = 5.19 Hz, 1H), 7.29 - 7.43 (m, J = 8.49 Hz, 2H), 7.17 - 7.29 (m, 2H), 7.05 - 7.17 (m, J = 8.49 Hz, 2H), 5.76 (s, 2H).

[0219] 1-(4-(4-Chlorobenzyl)-4H-thieno[3-b]pyrrole-5-carbonyl)-N-isopropylpiperidine-4-carboxamide (NCGC2955). To CH 2 Cl 2To a mixture of compound 4 (51.9 mg, 0.177 mmol, 1.0 equiv) in (1.7 mL) was added N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (37.5 mg, 0.19 mmol, 1.1 equiv), 1-hydroxybenzotriazole monohydrate (31.0 mg, 0.204 mmol, 1.15 equiv) and diisopropylethylamine (123 μL, 0.71 mmol, 4 equiv). After stirring for 5 minutes at room temperature, a solution of TFA salt 2 (60.7 mg, 0.21 mmol, 1.2 equiv) in CH 2 Cl 2 (0.2 mL, plus 0.2 mL rinse) was added and the reaction was stirred at room temperature. After 24 hours, the reaction was diluted with CH 2 Cl 2 and washed with 5% aqueous HCl. The aqueous layer was extracted with EtOAc (3 x 10 mL). The combined organic layers were dried over anhydrous MgSO 4 and concentrated in vacuo. Purification by flash chromatography (0 to 100% EtOAc / hexanes) afforded 48.9 mg of white solid NCGC2955 in 62% yield. 1 H NMR (500 MHz, CDCl 3 ) δ 7.24 (d, J = 8.33 Hz, 2H), 7.18 (d, J = 5.19 Hz, 1H), 7.08 (d, J = 8.33 Hz, 2H), 6.83 (d, J = 5.19 Hz, 1H), 6.58 (s, 1H), 5.43 (s, 2H), 5.19 (d, J = 8.02 Hz, 1H), 4.44 (d, J = 12.89 Hz, 2H), 4.08 (qd, J = 6.77, 13.34 Hz, 1H), 2.92 (t, J = 12.34 Hz, 2H), 2.24 (tt, J = 3.62, 11.08 Hz, 1H), 1.80 (d, J = 14.93 Hz, 2H), 1.43 - 1.63 (m, 6H), 1.15 (d, J = 6.60 Hz, 6H).

[0220] Synthesis of NFU1827: 4-(4-methoxyphenyl)-7-thia-2,5-diazatricyclo[6.4.0.02,6]dodeca-1(8),3,5,9,11-pentaene-10-carboxylic acid (5). 2-Aminobenzothiazole-6-carboxylic acid (2.0 g, 10.30 mmol, 1.0 equiv) and 2-bromo-4'-methoxyacetophenone (2.60 g, 11.33 mmol, 1.1 equiv) were charged into a pressure bottle and suspended in 2-methoxyethanol (50 mL). The suspension was heated to 40 °C for 120 h and then at 140 °C for 19 h. The reaction was allowed to cool to room temperature and concentrated in vacuo. The crude compound was purified by flash column chromatography (5 to 75% EtOAc / hexane) to afford 1.83 g of compound 6 in 55% yield. 1 H NMR (500 MHz, DMSO-d6) δ 13.17 (broad s, 1H), 8.69 (s, 1H), 8.63 (d, J = 1.2 Hz, 1H), 8.10 (dd, J = 1.6, 8.3 Hz, 1H), 8.01 (d, J = 8.4 Hz, 1H), 7.78 (d, J = 8.7 Hz, 1H), 6.99 (d, J = 8.8 Hz, 1H), 3.77 (s, 3H).

[0221] N-[3-(4-ethylpiperazin-1-yl)propyl]-4-(4-methoxyphenyl)-7-thia-2,5-diazatricyclo[6.4.0.02,6]dodeca-1(8),3,5,9,11-pentaene-10-carboxamide (NFU1827). Compound 5 (100 mg, 0.308 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (118.2 mg, 0.617 mmol), and 1-hydroxybenzotriazole hydrate (83.3 mg, 0.617 mmol) were dissolved in DMF and stirred at room temperature for 1 h. The reaction was then treated with 3-(4-ethylpiperazin-1-yl)propan-1-amine (0.2269 mL, 211.2 mg, 1.23 mmol) and stirred at room temperature for 18 h. The reaction was then poured into H 2 2O (20 mL). It was extracted with EtOAc (3 x 20 mL). The combined extracts were washed with H 2 2O (4 x 20 mL), followed by brine (1 x 20 mL). The organic layer was dried (Na 2 2SO 4) Filtered and the solvent removed in vacuo. The crude compound was purified by flash column chromatography using EtOAc / hexane (gradient elution from 90:10 to 0:100) to afford 60 mg of NFU1827 in 41% yield. 1H NMR (500 MHz, CDCl3) δ 8.71 (s, 1H), 8.21 (s, 1H), 7.99 (d, J = 8.4 Hz, 1H), 7.91 (s, 1H), 7.80 (d, J = 8.7 Hz, 2H), 7.62 (d, J = 8.4 Hz, 1H), 6.96 (d, J = 8.7 Hz, 2H), 3.85 (s, 3H), 3.60 (q, J = 5.2 Hz, 2H), 2.63 (t, J = 5.5 Hz, 2H), 2.46 (q, J = 7.3 Hz, 2H), 1.82 (m, 2H), 1.78 (broad s, 8H), 1.10 (t, J = 7.2 Hz, 3H). 13C NMR (126 MHz, CDCl3) δ 162.72, 156.34, 145.14, 130.89, 128.49, 127.24, 123.49, 123.25, 122.78, 120.26, 111.13, 109.09, 102.92, 57.32, 55.69, 52.27, 50.25, 49.91, 49.39, 38.42, 26.62, 20.55, 17.98, 11.12.C 26 H 31 N 5 O 2 S[M+H] + The HRMS (ESI + ) m / z calculated for 478.2272, found 478.2271.

[0222] Cells: Human foreskin fibroblasts (HFFs), passages 12 - 16; (ATCC, CRL-2088) were grown at 37 °C in a 5% CO 2 incubator in Dulbecco's modified Eagle medium (DMEM) (Gibco, Carlsbad, CA) containing 10% fetal bovine serum (FBS). Mouse embryonic fibroblasts (MEF, ATCC, CRL-1658) were infected with murine CMV. Vero cells were obtained from the laboratory of Dr. Gary Hayward at the Johns Hopkins University School of Medicine.

[0223] Cytotoxicity assay: The MTT assay was performed according to the manufacturer's instructions (Sigma-Aldrich). Uninfected cells were treated with the compound for 72 hours or 10 days (the same time points as in the antiviral assay), and 20 μL / well of MTT [3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide, 5 mg / mL phosphate-buffered saline (PBS) solution] was added to each well. After shaking at 150 rpm for 5 minutes, the plates were incubated at 37 °C for 2 - 3 hours. The yellow solution was converted to dark blue formazan by the mitochondrial dehydrogenase of live cells and quantified by measuring the absorbance at 560 nm. The formazan was quantified by measuring the absorbance at 560 nm.

[0224] Virus and antiviral assays: A recombinant pp28 luciferase Towne HCMV strain expressing luciferase under the control of the UL99 (pp28) late promoter was reported, providing a highly sensitive and reproducible reporter gene for drug screening. 20 A ganciclovir-resistant HCMV pp28 luciferase containing the C607Y mutation in UL97 was reported. 37 The HCMV TB40 strain was obtained from ATCC (VR-1578). Clinical isolates of human herpesvirus type 1 and 2 (HSV1, HSV2) were collected from the Johns Hopkins Microbiology Laboratory without any identifiers that could link them to specific patients. The mouse CMV (MCMV) Smith strain (ATCC VR-1399) was used for MEF infection.

[0225] Luciferase activity: As described previously, cell lysates were collected at 72 hpi, and the luciferase activity was determined using the Glomax Multi+ detection system (Promega, Madison, WI). 20 .

[0226] Plaque reduction assay: HFFs were seeded into 12-well plates (2 x 10 5 cells / well) and infected with HCMV TB40 at approximately 100 plaques / well. After 90 minutes, the medium was aspirated, and DMEM containing 0.5% carboxymethyl cellulose (CMC), 4% fetal bovine serum (FBS), and the drug was added to two replicate wells. After incubation at 37 °C for 10 - 12 days, the overlay was removed, and the plaques were counted after crystal violet staining. For HSV1 and HSV2 replication in Vero cells, the adsorption time was 60 minutes, and the plaques were counted 36 hours later. The plaque assay in MEF was completed after 3 days.

[0227] Inhibition of Epstein-Barr virus (EBV) replication: Latently EBV-infected Akata cells (0.5 x 10 6 / well in a 24-well plate) were induced with 100 μg / ml goat anti-human IgG (Sigma). Immediately after adding human IgG, the compounds and GCV control were added to three wells. Cells were harvested for DNA quantification 48 hours after lytic induction. Cell DNA was purified using the Wizard SV Genomic kit (Promega, Madison, WI). The effects of these compounds on EBV lytic induction were analyzed by real-time PCR of the EBV DNA polymerase gene BALF5 38 .

[0228] Addition and removal assays: These assays were performed to identify the stage during HCMV replication at which the compounds had the highest activity. In the addition group, the compounds were added to infected HFFs at 0, 6, 24, and 48 hpi, and luciferase activity was measured at 72 hpi. In the removal group, the compounds were added immediately after virus infection and then removed at 0, 6, 24, and 48 hours; luciferase was assayed at 72 hpi.

[0229] DNA isolation and real-time quantification (qPCR): Total DNA was isolated from uninfected controls and HCMV-infected HFFs using the Wizard SV Genomic DNA Isolation kit (Promega, Madison, WI). To determine the viral load in the supernatant, total DNA was isolated from the supernatant using automated DNA extraction on a BioRobot M48 instrument (Qiagen, Valencia, CA). A 151-bp US17 real-time PCR assay targeting a highly conserved US17 region from the CMV genome was used 39 . The primers and probe for US17 were:

[0230] Forward 5'-GCGTGCTTTTTAGCCTCTGCA-3' (SEQ ID NO:1),

[0231] Reverse 5'-AAAAGTTTGTGCCCCAACGGTA-3' (SEQ ID NO:2), and

[0232] US17 probe FAM 5'-TGATCGGGCGTTATCGCGTTCT-3' (SEQ ID NO:3).

[0233] HCMV Entry and Indirect Immunofluorescence Assay: MLS8969, GCV, and heparin controls were used to determine the inhibition of HCMV entry. Compounds were diluted in serum-free medium and added to HFFs seeded on coverslips 24 hours prior to infection. After 2 hours of infection (MOI 1 or 0.1 PFU / cell), cells were fixed with 100% chilled methanol and blocked with phosphate-buffered saline (PBS), 5% serum, and 0.3% Triton X-100 for 1 hour. Cells were then incubated with a 1:50 mouse monoclonal anti-pp65 antibody (Vector Laboratories, Burlingame, CA) in a humidified chamber at 37 °C for 1 hour, washed three times with TBST (0.1%), incubated with a 1:500 rhodamine-conjugated anti-mouse IgG (Sigma) in a humidified chamber at 37 °C for 1 hour, and washed with TBST (0.1%). Prior to visualization with a Zeiss ZI fluorescence microscope, a drop of mounting oil containing DAPI (Santa Cruz Biotechnology, Santa Cruz, CA) was added to the coverslip. Images were taken at 40× magnification.

[0234] SDS Polyacrylamide Gel Electrophoresis and Immunoblot Analysis: Cell lysates containing equal amounts of protein were mixed with an equal volume of sample buffer (125 mM Tris-HCL, pH 6.8, 4% SDS, 20% glycerol, and 5% β-mercaptoethanol) and boiled at 100 °C for 10 minutes. Denatured proteins were resolved in a Tris-glycine polyacrylamide gel (8–10%) and transferred to a polyvinylidene difluoride (PVDF) membrane (Bio-Rad Laboratories, Hercules, CA) by electroblotting. Membranes were incubated in blocking solution [5% w / v non-fat dry milk and PBS containing 0.1% Tween-20 (PBST)] for 1 hour, washed with PBST, and incubated with primary antibody overnight at 4 °C. Membranes were washed with PBST and incubated with horseradish peroxidase-conjugated secondary antibody in PBST at room temperature for 1 hour. After washing with PBST, protein bands were visualized by chemiluminescence using SuperSignal West Dura and Pico reagents (Pierce Chemical, Rockford, IL).

[0235] Antibodies: The following antibodies were used - mouse monoclonal anti-HCMV IE1 and IE2 (MAB810, Millipore, Billerica, MA), mouse monoclonal anti-HCMV UL83 (pp65, Vector Laboratories, Burlingame, CA), mouse monoclonal antibody HCMV UL44, mouse monoclonal anti-HCMV UL84, and mouse anti-actin anti-mouse IgG (Santa Cruz Biotechnology, Santa Cruz, CA). Horseradish peroxidase (HRP)-conjugated anti-mouse IgG was from GE Healthcare (Waukesha, WI). Horseradish peroxidase (HRP)-conjugated anti-rabbit IgG was from Cell Signaling (Beverly, MA).

[0236] Drug Combinations and Analysis: These experiments were conducted as previously reported 40 . The combination of GCV and each compound was tested using the pp28 luciferase assay. Briefly, 2x10 6 HFF / well were seeded in 96-well plates and infected with the pp28 luciferase Towne HCMV strain (MOI = 1). First, dose-response curves were generated separately for each drug alone to determine its EC 50 . Then, the drugs were combined at twice their EC 50 , diluted in DMEM with 4% FBS, then serially diluted and added together after infection. The luciferase activity of the combinations and each drug alone was quantified at 72 hpi. The combination of letermovir and each novel compound was tested using the plaque assay with TB40. The same principle applied to the assay of the drug combination effect, and the plaques were counted at 8 dpi. The Bliss model was used to calculate the effect of each drug combination on pp28 luciferase activity and plaque reduction. In this model, the drug combination represents the product of two probabilistically independent events, as described by the following equation 41 :

[0237]

[0238] where D is the drug concentration, m is the slope, and EC 50 is the effective concentration that results in 50% virus inhibition. The combined effect of two inhibitors (Fu, fraction unaffected) was calculated as the product of the individual effects of the two inhibitors Fu 1 and Fu 2 . If the ratio of the observed fold inhibition divided by the expected fold inhibition is greater than 1, the compounds are synergistic. If the ratio is less than 1, the combination is considered antagonistic, and if it equals 1, the combination is additive.

[0239] Statistical analysis: Student's t-test was performed using Sigmaplot (Systat Software, Inc., San Jose, CA) and GraphPad Prism (GraphPad Software, Inc., La Jolla, CA). One-way ANOVA was used to compare all sample groups with the control group. The P value was adjusted for multiple comparisons. In all figures, the following convention was followed. * indicates p value < 0.05, ** indicates p value < 0.01, *** indicates p value < 0.001. The Curve Fitting Toolbox, MATLAB software (v7.10), MathWorks (Natick, MA) was used to determine EC 50 and CC 50 values.

[0240] Example 2

[0241] High-throughput screening for HCMV inhibitors: pp28-HCMV luciferase has been shown to provide highly reproducible and sensitive results for inhibiting HCMV with compounds, for the MLSMR collection of 370,000 compounds, the NCGC diversity collection of 65,000 compounds, and the NCGC drug collection of approved and investigational drugs. Screening was performed using 2 compound doses pre-plated before infection, and luciferase activity of the lysates was detected 72 hours post-infection (hpi). Using the published compound dose-response curve algorithm 21 , a total of 2215 HCMV luciferase inhibitors were selected. Secondary screening was performed to remove compounds that showed toxicity in human foreskin fibroblasts (HFF), yielding 847 confirmed non-toxic HCMV inhibitors ( Figure 1 ).

[0242] HCMV inhibition was measured by luciferase activity (72 hpi) of the pp28 recombinant Towne and pp28 ganciclovir-resistant Towne strains of HCMV and plaque reduction (10 days) using HCMV Towne or TB40. Viral protein expression was determined by Western blot analysis. A colorimetric MTT assay was performed in uninfected HFF to determine cytotoxicity in parallel with the infectivity assay, i.e., at 72 hours and 10 days, and expressed as CC 50 . All 5 compounds were active against all HCMV strains used at sub-μM to low μM concentrations. Based on plaque reduction, MLS8969 had the best activity against HCMV, followed by MLS8091, MLS8554, NCGC2955, and NFU1827 (Table 1).

[0243] Table 1. Anti-HCMV activity, toxicity, and slope of 5 compounds against different HCMV strains

[0244]

[0245] * = Pretreatment, followed by plaque reduction assay

[0246] ** = Plaque reduction assay

[0247] Calculate the slope parameter (similar to the Hill coefficient) for each compound. A slope of 1 indicates one binding site or non - cooperativity. Positive cooperativity exists when binding at one site increases the ligand - binding affinity at another site (slope > 1). If binding at one site decreases the ligand affinity at another site, the compound exhibits negative cooperativity (slope < 1). For other chronic viral infections such as HIV, the slope has been found to be not only an important factor in differentiating drug classes and known mechanisms of action, but also helpful in reducing antiviral activity and even the EC of drug - resistant viral mutants 22, 23 50 remains unchanged. Based on the plaque reduction assay, the slope of all compounds except MLS8969 was 1.

[0248] Example 3

[0249] Inhibition of other herpesviruses: These five compounds were tested for inhibition of herpes simplex virus 1 and 2 (HSV1, HSV2), Epstein - Barr virus (EBV), and murine CMV (MCMV). The plaque reduction assay was used for HSV1, HSV2, and MCMV. After IgG induction, inhibition of EBV replication in AKATA cells was detected by real - time PCR. Only MLS8554 was active against HSV1, HSV2, and MCMV (Table 2).

[0250] Table 2. Activity of five compounds against other herpesviruses; NA - no activity; CI - clinical isolate

[0251]

[0252] NCGC2955 was active against MCMV (EC 50 was 1 μM, Table 2), but toxicity to MEF was observed at 10 μM and higher concentrations. NFU1827 showed activity against EBV (EC50 - 1.92 ± 0.1 μM). The other compounds had specific activity against HCMV but did not inhibit HSV1 or MCMV.

[0253] Example 4

[0254] Time of HCMV inhibition: Addition and removal experiments were performed. Compounds were added or removed at 0, 6, 12, 24, 36, and 48 hpi, and luciferase activity was measured at 72 hpi. Based on these experiments, each compound had a specific time of maximum activity, and the time based on maximum virus inhibition was recorded as follows.

[0255] Example 5

[0256] Inhibition of HCMV entry: Compound MLS8969 was first tested using a luciferase assay at 72 hpi after infection at an MOI of 1 PFU / cell, but a minimal decrease in luciferase enzyme activity was observed ( Figure 3 A). However, plaque assays showed good anti-HCMV activity of MLS8969 ( Figure 3 B).

[0257] Since in high-throughput screening, HFFs were treated with the compound before infection (pretreatment), and our plaque assays (addition of MLS8969 after infection) showed an EC 50 of 0.12 ± 0.02 μM (Table 1), we suspected that the lack of luciferase activity inhibition might represent an MOI-dependent phenomenon, thus suggesting entry inhibition 24 . Therefore, experiments were performed using MOIs of 0.01, 0.1, and 1 PFU / cell ( Figure 3 C), and MLS8969 was added before infection. An MOI-dependent inhibition of luciferase activity was observed ( Figure 3 C). In plaque reduction assays, pretreatment with MLS8969 also inhibited TB40 and ganciclovir-resistant HCMV Towne ( Figure 3 D, 3E). Western blot analysis confirmed that these data did not show inhibition of viral protein expression under post-treatment or pretreatment at an MOI of 1 PFU / cell, while pretreatment after infection at a low MOI (0.01) led to reduced protein levels of IE1 / 2, pp65, UL84, and UL44 ( Figure 3 F). Further evidence that MLS8969 is an entry inhibitor was obtained by indirect immunofluorescence assay (IFA) of HCMV pp65 ( Figure 4 ). MLS8969 inhibited HCMV entry only at an MOI of 0.1; ganciclovir did not affect pp65 nuclear staining regardless of the MOI ( Figure 4 ). Heparin (positive control) inhibited HCMV entry.

[0258] Example 6

[0259] Inhibitors of the immediate early-to early stage of HCMV replication: Two compounds showed anti-HCMV activity (NFU1827 and MLS8554) at 0 to 24 hpi. NFU1827 was active only against HCMV and GCV-R HCMV, with EC 50 values of 0.85 ± 0.1 and 0.84 ± 0.1 μM, respectively (Table 1, Figure 5 A and B), but was not active against MCMV or HSV1 (Table 2). The add-back and removal experiments indicated the immediate early time of activity. When added to infected cells after 24 hours, it lost its ability to inhibit HCMV. Removal of NFU1827 after 6 hours had achieved significant HCMV inhibition ( Figure 5 C&D). NFU1827 reduced the levels of IE1 and IE2 ( Figure 5 E). According to its immediate early activity, viral DNA replication and DNA production in the supernatant were significantly reduced ( Figure 5 F&G).

[0260] MLS8554 was active in all antiviral assays against HCMV, GCV-R HCMV, MCMV, and HSV1 ( Figure 6 , Table 1 and Table 2). Its maximum activity was determined at the immediate early time after infection. When added at 24 hpi or later, its activity was significantly reduced, and when removed at 6 hours, HCMV inhibition was nearly complete ( Figure 6 C&D). Western blot analysis at 72 hpi showed reduced expression of IE2, UL44, UL84, and pp65 ( Figure 6 E). Compared with NFU1827, which reduced both IE1 and IE2, MLS8554 mainly reduced IE2 expression. Both viral DNA replication and DNA supernatant production were reduced ( Figure 6 F&G).

[0261] Example 7

[0262] Inhibitors of the early-to late stage of HCMV replication: MLS8091 showed a dose response against HCMV and GCV-resistant HCMV, and EC 50 values of 0.39 ± 0.5 and 0.26 ± 0.02 μM, respectively (Table 1, Figure 7 ), but was not active against HSV1 or MCMV (Table 2). In the add-back experiment, its activity time overlapped with that of GCV, but in the removal experiment, MLS8091 showed an earlier effect ( Figure 7 C&D). Inhibition of all viral proteins was observed at 72 hpi ( Figure 7E). Although the impact on DNA yield was not significantly different from GCV, the inhibition of viral DNA replication was greater than GCV ( Figure 7 F and 7G).

[0263] NCGC2955 inhibits HCMV and GCV-R HCMV ( Figure 8 A and 8B). For this compound, the plaque assay was preferred because it did not show a full dose response using the luciferase assay. In the addition assay, the active time of NCGC2955 overlapped with GCV, but in the removal assay, NCGC2955 showed longer activity than GCV ( Figure 8 C & D). At 72 hpi, there was a significant decrease in the expression of viral proteins including IE1 and IE2 ( Figure 8 E). Although viral progeny was inhibited and viral protein expression was inhibited by the plaque assay, the impact of NCGC2955 on viral DNA replication and viral DNA yield was small ( Figure 8 F and 8G), indicating inhibition by a mechanism not involving the DNA replication machinery.

[0264] Example 8

[0265] Drug combination patterns with GCV and letermovir: The effects of each compound used alone or in combination with GCV or letermovir were tested in HCMV-infected HFFs. For all five reagents, additive or mildly synergistic effects with GCV ( Figure 9 ) or letermovir ( Figure 10 ) were observed. A ratio of the observed inhibition fold divided by the expected inhibition fold > 1 was considered a drug synergistic effect, < 1 was considered a drug antagonistic effect, and = 1 was considered an additive effect. The Bliss coefficients of the drug combinations are shown in Table 3. For letermovir and its combinations with each of the five compounds, EC 50 was determined based on the plaque assay. The combination of GCV and 2955 was tested by the plaque assay. All other combinations and Bliss coefficients were determined using pp28-luciferase. All Bliss coefficients were close to 1, indicating an additive effect between the compounds.

[0266] Table 3. Bliss coefficients of drug combinations

[0267]

[0268] Example 9

[0269] Discussion and conclusions: We report the results of the largest HTS to date using our pp28 luciferase HCMV against HCMV inhibitors. This reporter virus showed high sensitivity and reproducibility in this screen 20。Its advantage is that pp28 encoded by HCMV is a true late gene, so its activation represents that the HCMV replication approaches a complete cycle.

[0270] The screening campaign may lead to false positive hits that require careful validation. Our combined approach using multiple HCMV strains and multiple detailed antiviral assays led to the identification and characterization of five different hits, each with its unique features. For the five hits, MLS8969 inhibits HCMV entry, MLS8554 and NFU1827 inhibit the immediate-early phase of HCMV replication (but have different effects on other herpesviruses), MLS8091 and NCGC2955 are active in the early-late phase, but NCGC2955 does not strongly inhibit viral DNA replication. When used in combination with GCV or maribavir, all five compounds showed additive effects or slight synergy. No antagonistic effects were found in all drug combination experiments. In addition, the five compounds are active against GCV-resistant HCMV, indicating that their mechanisms are different from that of the viral DNA polymerase. Future studies should provide in-depth understanding of the mechanisms of action of the five compounds. Based on the data provided here, we expect that each compound has a unique HCMV inhibition mechanism.

[0271] Screening was performed using a library containing a limited number of small molecules, different HCMV strains, cell types, and treatment times. Based on fluorescence intensity, multiple kinase inhibitors blocked HCMV replication. Three recombinant HCMVs carrying enhanced yellow fluorescent protein (EYFP) fused to immediate-early viral protein 2 (IE-2), ppUL32 (pp150), and ppUL83 (pp65) were applied to a cell kinase library (with 80 compounds) as a pilot screening. 25 。The screening used a fixed drug concentration of 10 μM at the time of infection, and a limited number of hits had no obvious cytotoxicity. Reporter cell lines have been generated to screen for anti-HCMV compounds. 26、27 。In one approach using a luciferase reporter cell line, the promoter provides IE protein activation; thus, compounds that inhibit HCMV at a later stage of infection cannot be evaluated. 27 。The Gray Kinase Inhibitor library of 187 compounds was successfully screened with HCMV strain AD169, and the cells were stained with antibodies to detect the HCMV antigen pp28. Three kinase inhibitors were identified as inhibitors of IE2 production. 28 。A recent drug repurposing screening used a spectral set of 2,560 compounds from Micro Discovery System. 29,30. Mercorelli used mechanism-based screening to identify compounds that interfere with IE2 transactivation activity. The phenotypic assay measured the expression of EGFP fused to IE2 31 or UL99 32 . The major success of the HTS was the identification of letermovir (AIC246) from the screening of a compound library 33 , which was shown to have a novel mechanism of action against HCMV in subsequent studies 34 .

[0272] In summary, our study provides several novel compounds that inhibit HCMV replication at sub-μM to low μM concentrations. Each of these compounds is a good candidate for future mechanistic studies and drug development.

[0273] All references cited herein (including publications, patent applications, and patents) are incorporated herein by reference to the same extent as if each reference were individually incorporated by reference and set forth in its entirety herein. The description of the present disclosure is such that the terms "a", "an", "the", and similar expressions used in the context (especially in the context of the appended claims) should be construed to cover the singular and the plural unless otherwise indicated or the context clearly dictates otherwise. The terms "comprising", "having", "including", and "containing" should be construed as open-ended terms (i.e., meaning "including, but not limited to") unless otherwise noted. Unless otherwise indicated herein, the reference to a numerical range herein is merely a shorthand method of indicating each individual value falling within the range, and each individual value is incorporated into the specification as if it were individually recited herein. All methods described herein may be performed in any suitable order unless otherwise indicated herein or clearly contradicted by the context. Unless otherwise noted, the use of any and all examples or exemplary language (e.g., "such as") provided herein is merely for the purpose of better illuminating the present disclosure and is not a limitation on the scope of the disclosure. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the present disclosure.

[0274] The present disclosure includes all modifications and equivalents of the subject matter recited in the appended claims as permitted by applicable law. In addition, unless otherwise indicated herein or clearly contradicted by the context, the present disclosure covers any combination of all possible variations of the above elements.

[0275] List of References

[0276] 1. Staras, SA; Dollard, SC; Radford, KW; Flanders, WD; Pass, RF; Cannon, MJ. Serological evaluation of cytomegalovirus infection in the United States, 1988-1994. Clin. Infect. Dis 2006, 43, 1143-1151.

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Claims

1. A method for inhibiting human herpesvirus in a subject in need thereof, comprising administering to the subject a compound of formula I or a pharmaceutical composition thereof or a salt, solvate or stereoisomer thereof, said pharmaceutical composition comprising a pharmaceutically acceptable carrier: Wherein, R 1 is an optionally substituted cycloalkyl, heterocycloalkyl, aryl or heteroaryl having 1 to 10 carbon atoms and substituted by one or more amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidine, acyl guanidine, and is linked by a C 1 -C 3 alkyl linkage.

2. The method according to claim 1, characterized in that, the compound is selected from the group consisting of:

3. A method for inhibiting human herpesvirus in a subject in need thereof, comprising administering to the subject a compound of formula II or a pharmaceutical composition thereof or a salt, solvate or stereoisomer thereof, said pharmaceutical composition comprising a pharmaceutically acceptable carrier: Wherein, R 1 is H, and R 2 independently is H or an alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl optionally substituted by one or more alkyl, amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidine, acyl guanidine, having 1 to 10 carbon atoms and attached through a C 1 -C 3 alkyl linkage; or R 1 and R 2 together form an optionally C substituted by one or more amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidine, acyl guanidine 1 -C 6 heteroalkyl or C 1 -C 6 heteroaryl, having 1 to 10 carbon atoms; and R 3 is H or an optionally cycloalkyl, heteroalkyl, aryl or heteroaryl substituted by one or more halogen, CF 3 , CN and NO 2 moieties.

4. The method according to claim 3, wherein, R 2 is optionally C substituted by one or more alkyl, amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidine, acyl guanidine 1 -C 6 heterocycloalkyl or C 1 -C 6 heteroaryl, having 1 to 10 carbon atoms and connected by C 1 -C 3 alkyl linkage.

5. The method according to claim 3 or 4, wherein, the compound is selected from the group consisting of:

6. A method for inhibiting human herpesvirus in a subject in need thereof, comprising administering to the subject a compound of the following formula or a salt, solvate or stereoisomer thereof:

7. A method for inhibiting human herpesvirus in a subject in need thereof, comprising administering a compound of formula IV or a pharmaceutical composition thereof or a salt, solvate or stereoisomer thereof, said pharmaceutical composition comprising a pharmaceutically acceptable carrier:

8. A method for inhibiting human herpesvirus in a subject in need thereof, comprising administering to the subject a compound of formula V or a pharmaceutical composition thereof or a salt, solvate or stereoisomer thereof, said pharmaceutical composition comprising a pharmaceutically acceptable carrier: Wherein, R 1 is hydrogen, a C1-C6 straight or branched alkyl group, or a C3-C6 cycloalkyl group; R 2 is an unsubstituted phenyl group or a phenyl group substituted by R 3 or OR 3 wherein R 3 is hydrogen, a C1-C6 straight or branched alkyl group, or a C3-C6 cycloalkyl group.

9. The method according to claim 8, wherein, R 1 is a C1-C4 straight or branched alkyl group; and R 2 is a phenyl group substituted by OR 3 wherein R 3 is a C1-C4 straight or branched alkyl group.

10. The compound according to claim 9, wherein, the compound is 11. The method according to any one of claims 1 to 10, wherein, the herpesvirus is selected from human herpesvirus type 1 (HHV-1), human herpesvirus type 2 (HHV2), human herpesvirus type 3 (HHV3), human herpesvirus type 4 (HHV4), human herpesvirus type 5 (HHV5), human herpesvirus type 6 (HHV6), human herpesvirus type 7 (HHV7) and human herpesvirus type 8 (HHV8).

12. The method according to claim 11, wherein, the herpesvirus is HHV-5.

13. The method according to claim 12, wherein, HHV-5 is resistant to ganciclovir.

14. The method according to any one of claims 1-13, said method further comprising administering more than one bioactive agent.

15. The method according to any one of claims 1-14, said method further comprising administering ganciclovir or letermovir.

16. A pharmaceutical composition comprising a compound of formula I or a salt, solvate or stereoisomer thereof and a pharmaceutically acceptable carrier: Wherein, R 1 is an optionally substituted cycloalkyl, heterocycloalkyl, aryl or heteroaryl, substituted by one or more amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidine or acyl guanidine, having 1 to 10 carbon atoms and linked through a C 1 -C 3 alkyl linkage.

17. The pharmaceutical composition according to claim 16, wherein, the compound is selected from the group consisting of:

18. A pharmaceutical composition comprising a compound of formula II: Wherein, R 1 is H, and R 2 independently is H or an alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl optionally substituted by one or more alkyl, amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidine, acyl guanidine, having 1 to 10 carbon atoms and attached via C 1 -C 3 alkyl linkage; or R 1 and R 2 together form a C 1 -C 6 heterocycloalkyl or a C 1 -C 6 heteroaryl, having 1 to 10 carbon atoms and optionally substituted by one or more amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidine, acyl guanidine; and R 3 is H or a cycloalkyl, heterocycloalkyl, aryl or heteroaryl optionally substituted by one or more halogen, CF 3 , CN and NO 2 moieties.

19. The pharmaceutical composition according to claim 18, wherein, R 2 is optionally C substituted by one or more alkyl, amidino, guanidino, phosphate, sulfate, tetrazole, 3-hydroxyisoxazole, secondary amide, sulfonamide, sulfonylurea, acyl amidine, acyl guanidine 1 -C 6 heterocycloalkyl or C 1 -C 6 heteroaryl, having 1 to 10 carbon atoms and linked through C 1 -C 3 alkyl linkage.

20. The pharmaceutical composition according to claim 19, wherein, the compound is selected from the group consisting of:

21. A pharmaceutical composition comprising a compound of the following formula or a salt, solvate or stereoisomer thereof and a pharmaceutically acceptable carrier:

22. A pharmaceutical composition comprising a compound of formula IV or a salt, solvate or stereoisomer thereof and a pharmaceutically acceptable carrier:

23. A pharmaceutical composition comprising a compound of formula V or a salt, solvate or stereoisomer thereof and a pharmaceutically acceptable carrier: wherein, R 1 is hydrogen, a C1-C6 straight or branched alkyl group, or a C3-C6 cycloalkyl group; and R 2 is an unsubstituted or R 3 or OR 3 substituted phenyl group, wherein R 3 is hydrogen, a C1-C6 straight or branched alkyl group or a C3-C6 cycloalkyl group.

24. The pharmaceutical composition according to claim 23, wherein, R 1 is a C1-C4 straight or branched alkyl group; and R 2 is a phenyl group substituted by OR 3 , wherein R 3 is a C1-C4 straight-chain or branched alkyl group.

25. The pharmaceutical composition according to claim 24, wherein, the compound is 26. The pharmaceutical composition according to any one of claims 16 - 25, further comprising more than one bioactive agent.

27. The pharmaceutical composition according to claim 26, the composition further comprising ganciclovir or letermovir.

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