Antiviral 1,3-di-oxo-indene compounds

Abstract

The present disclosure describes a compound of formula (I): [I] (I) as described herein is provided together with a pharmaceutically acceptable salt, a pharmaceutical composition containing such compound, and a method of using such compound, salt and composition to treat a viral infection.

Description

Technology Field

[0001] The present invention relates to an inhibitor of picornaviruses, including coxsackievirus, enterovirus, echovirus, poliovirus, and rhinovirus, and to poliomyelitis, paralysis, acute hemorrhagic conjunctivitis, viral meningitis, hand-foot-and-mouth disease, vesicular disease, hepatitis A, myositis, myocarditis, pancreatitis, diabetes mellitus, epidemic myalgia, encephalitis, the common cold, herpangina, foot-and-mouth disease, asthma, chronic obstructive pulmonary disease, pneumonia, The present invention relates to novel 1,3-dioxoindene compounds useful for treating viral infections, including sinusitis or otitis media. The present invention provides novel tetracyclic pyridone compounds as disclosed herein, pharmaceutical compositions containing such compounds, and methods for using these compounds and compositions in the treatment and prevention of viral diseases. Background Technology

[0002] Picornaviridae are non-enveloped, positive single-stranded RNA viruses with RNA genomes 7.2 to 8.5 kb in length. These viruses are very small, spherical in shape, and about 22 to 30 nm in size, and were first identified a long time ago. Among the viruses belonging to the family Picornaviridae are rhinovirus, poliovirus, enteroviruses including Coxsackievirus A, Coxsackievirus B, and echovirus, and hepatitis A virus.

[0003] Diseases caused by picornaviruses range from respiratory diseases to digestive, circulatory, and skin diseases; examples include polio, paralysis, acute hemorrhagic conjunctivitis, viral meningitis, hand, foot, and mouth disease, vesicular disease, hepatitis A, myositis, myocarditis, pancreatitis, diabetes mellitus, epidemic myalgia, encephalitis, the common cold, herpangina, and foot-and-mouth disease. However, no cure exists for these diseases. Most drugs under development are uncoating inhibitors. Viruses belonging to the Picornaviridae family cause a variety of diseases, including the aforementioned respiratory diseases, which lead to hygiene, social, and economic problems. Picornaviruses are the primary causative agents of waterborne diseases. Because they are highly stable and difficult to disinfect, RNA viruses continuously cause related diseases.

[0004] Human rhinovirus (hRV) has recently been associated with most asthma exacerbations and is known to be present in the bronchial tissues of many stable asthma patients. Comparisons of bronchial mucosal biopsy specimens obtained from asthmatic and non-asthmatic patients have shown a significantly higher detection frequency of human rhinovirus in the lower respiratory tract of asthmatic patients compared to non-asthmatic patients. Furthermore, a correlation has been reported between the presence of human rhinovirus and the clinical severity of asthma. In addition, rhinovirus causes chronic obstructive pulmonary disease, pneumonia, sinusitis, and otitis media, as well as asthma.

[0005] Rhinovirus is a major cause of the common cold, while enterovirus-induced diseases include meningitis and respiratory infections. Extensive efforts to provide vaccines against poliovirus have significantly reduced the incidence of polio globally, yet cases of the disease are still being reported in Niger, Nigeria, Egypt, India, Pakistan, and Afghanistan. Hepatitis A is now under some degree of control thanks to vaccines against the hepatitis A virus. However, no vaccines have been developed to date for Coxsackievirus, Echovirus, or Rhinovirus.

[0006] In particular, Coxsackievirus B is a major cause of myocarditis, which can lead to idiopathic dilated cardiomyopathy requiring a heart transplant in severe cases.

[0007] Enviroxime derivatives are considered the most promising candidates for possessing broad anti-enteroviral and anti-rhinoviral activities. Enviroxime interferes with the synthesis of plus-strand RNA by binding to viral protein 3A, which is required for the formation of RNA intermediates during viral replication (Reference [Heinz BA and Vance LM: J Virol, 1995, 69(7), 4189-97]). However, in clinical studies, the compound was observed to have insignificant or minimal therapeutic effects, accompanied by poor pharmacokinetics and the incidental detection of unwanted side effects (Reference [Miller FD et al.: Antimicrob Agents Chemother, 1985, 27(1), 102-6]).

[0008] The protease inhibitor AG 7088 has been developed based on knowledge of the microstructure and function of viral protease 2C. In cell cultures at nanomolar concentrations, AG 7088 is effective against 48 rhinovirus types, as well as Coxsackievirus A21, B3, Enterovirus 70, and Echovirus 11 (Reference [Pattick AK et al.: Antimicrobila Agents Chemother, 1999, 43(10), 2444-50]).

[0009] Thanks to the description of the molecular structure of viral capsids, preliminary conditions were obtained for the intentional design of capsid blockers, "WIN components" (Reference [Diana GD: Curr Med Chem 2003, 2, 1-12]). These inhibit the adsorption and / or non-coating of rhinoviruses and enteroviruses. Some WIN components have highly specific effects only on individual genera or virus types of picornaviruses. Other derivatives inhibit the replication of both rhinoviruses and enteroviruses. Arildone, dioxaril, and pirodavir belong to the WIN components, for example. These compounds have demonstrated very good antiviral effects in cell cultures. However, poor solubility (aryldone), low bioavailability (aryldone and disoxalil), rapid metabolism and elimination (dioxalil and WIN 54954), as well as side effects such as skin rash (WIN 54954), made clinical application impossible.

[0010] Pleconaril, a type of WIN component, has very good oral bioavailability and, after binding to the hydrophobe pocket within the viral capsid, inhibits the penetration of rhinovirus, echovirus, and coxsackievirus (Pevear DC et al.: Antimicrob Agents Chemother 1999, 43(9), 2109-15; McKinlay MA et al.: Annu Rev Microbiol 1992, 46, 635-54). Therefore, pleconaril is potentially effective against a broad spectrum of viral diseases ranging from the common cold to viral meningitis or myocarditis. Resistance has been observed against rhinovirus, enterovirus 71, and coxsackievirus B3 (Ledford RM et al.: J Virol 2004, 78(7), 3663-74; Groarke JM et al.: J Infect Dis 1999, 179(6), 1538-41). However, the proven therapeutic effect was not sufficient for the registration of pleconaril (Picovir, Viropharma, USA) as a treatment for rhinovirus infection in the United States. In March 2002, the corresponding application was rejected by the U.S. Food and Drug Administration (FDA) because the success rate of the therapy was too low and side effects were observed.

[0011] BTA-798 was found to have higher antiviral activity than pleconaril, as evaluated in vitro and in vivo using rhinovirus, and is currently undergoing clinical trials (Reference [Ryan, J. et al. Antiviral Res [18th Intl Conf Antiviral Res (April 11-14, Barcelona) 2005] 2005, 65(3): Abst LB-11]).

[0012] However, no antiviral drug approved for the treatment of enteroviruses or rhinoviruses has been developed to date. The need for new treatments and therapies for enteroviruses or rhinoviruses remains.

[0013] Up to the present invention, in-depth and thorough research on effective viral replication inhibitors (virtatatics) against picornaviruses, including Coxsackievirus, Enterovirus, Echovirus, Poliovirus, and Rhinovirus, has led to the discovery that novel 1,3-dioxoindene derivatives exhibit high inhibitory activity against picornaviruses, including Coxsackievirus, Enterovirus, Echovirus, Poliovirus, and Rhinovirus. The problem to be solved

[0014] The present invention provides a compound having antiviral activity. The present invention also provides a pharmaceutical composition containing the compound, as well as a method of using said compound and composition to inhibit viral replication or reactivation and to treat disease conditions associated with or caused by the virus. Further objects of the present invention are described in the following detailed description and examples. means of solving the problem

[0015] In one embodiment, the present invention provides a compound of formula (I):

[0016] [I]

[0017] In the above formula,

[0018] G 1 and G 2 One of them is selected from linear or branched C1-C5 alkyl; linear or branched C1-C5 alkyloxy; linear or branched C1-C5 haloalkyl; linear or branched C1-C5 haloalkyloxy; halo and 3-7-one-cycloalkyl; and G 1 and G 2 The other one is H; R 1The compound is selected from H and linear or branched C1-C5 alkyls; optionally, the compound is in a mirror image isomerically pure form. In another aspect, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of the compound of the present invention and one or more pharmaceutically acceptable carriers. In another aspect, the present invention provides a combination, particularly a pharmaceutical combination, comprising a therapeutically effective amount of the compound of the present invention and one or more therapeutic active agents. Effects of the invention

[0019] Since compounds represented by the chemical formula according to the present invention, which exist in a mutually equilibrium state, have low cytotoxicity as well as excellent antiviral activity against picornaviruses including Coxsackievirus, Enterovirus, Echovirus, Poliovirus, and Rhinovirus, they can be effectively used as pharmaceutical compositions for the prevention or treatment of viral diseases, such as polio, acute hemorrhagic conjunctivitis, viral meningitis, hand, foot, and mouth disease, blistering disease, hepatitis A, myositis, myocarditis, pancreatitis, diabetes, epidemic myalgia, encephalitis, influenza, herpangina, foot and mouth disease, asthma, chronic obstructive pulmonary disease, pneumonia, sinusitis, or otitis media. Specific details for implementing the invention

[0020] To interpret this specification, the following definitions will apply, and whenever appropriate, terms used in the singular will also include the plural.

[0021] Unless the context clearly indicates otherwise, the terms used herein have the following meanings:

[0022] As used herein, the term “subject” refers to an animal. In certain embodiments, the animal is a mammal. The subject also refers, for example, to primates (e.g., humans), cattle, sheep, goats, horses, dogs, cats, rabbits, rats, mice, fish, birds, etc. In certain embodiments, the subject is a human. As used herein, the term “patient” refers to a human subject. As used herein, the subject is “in need of treatment” if such subject would benefit biologically, medically, or in quality of life from such treatment.

[0023] As used herein, the terms “inhibition” or “inhibiting” refer to the reduction or suppression of a certain condition, symptom or disorder, or disease, or a significant decrease in the reference activity of a biological activity or process.

[0024] As used herein, the term “treating” or “treatment” of any disease or disorder refers, in one embodiment, to improving the disease or disorder (i.e., slowing down, stopping, or reducing the onset of the disease or at least one of its clinical symptoms). In another embodiment, “treating” or “treatment” refers to alleviating or improving at least one physical parameter, including one that is not identifiable by the patient. In another embodiment, “treating” or “treatment” refers to controlling the disease or disorder physically (e.g., stabilization of identifiable symptoms), physiologically (e.g., stabilization of physical parameters), or both. In yet another embodiment, “treating” or “treatment” refers to preventing or delaying the onset, onset, or progression of the disease or disorder.

[0025] As used herein, the singular forms (“a,” “an,” “the”) and similar terms used in the context of the invention (particularly in the context of the claims) shall be interpreted to include both singular and plural forms unless otherwise indicated herein or clearly contradictory in the context.

[0026] All methods described herein may be performed in any suitable order unless otherwise indicated herein or clearly contradictory in the context. The use of any examples or exemplary language (e.g., “e.g., for example”) provided herein is merely for better illustrating the invention and does not limit the scope of the invention as otherwise claimed.

[0027] "Halo" or "halogen" as used herein may be fluorine, chlorine, bromine, or iodine.

[0028] "C" used in this document 1-6 "alkyl" or "C1-C6alkyl" indicates a straight-chain or branched alkyl group having 1 to 6 carbon atoms. If a different number of carbon atoms, e.g., C4 or C3, is specified, the definition must be modified accordingly, e.g., "C 1-4 "Alkyl" will represent methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, and tert-butyl.

[0029] "C" used in this document 1-6 "Alkoxy" indicates a straight-chain or branched alkoxy (-O-alkyl) having 1 to 6 carbon atoms. If a different number of carbon atoms, e.g., C4 or C3, is specified, the definition must be modified accordingly, e.g., "C 1-4 "Alkoxy" will represent methoxy, ethoxy, propoxy, isopropoxy, butoxy, sec-butoxy, and tert-butoxy.

[0030] "C" used in this document 1-4 "Haloalkyl" or "C1-C4 haloalkyl" denotes a straight-chain or branched alkyl group having 1 to 4 carbon atoms, wherein at least one hydrogen is replaced by a halogen. The number of halogen substitutions may range from 1 to the number of hydrogen atoms on the unsubstituted alkyl group. If a different number of carbon atoms, e.g., C6 or C3, is specified, the definition must be corrected accordingly. Therefore, "C1-4 "Haloalkyl" will represent methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, and tert-butyl having at least one hydrogen substituted with a halogen, e.g., the halogen is fluorine: CF3CF2-, (CF3)2CH-, CH3-CF2-, CF3CF2-, CF3, CF2H-, CF3CF2CH(CF3)-, or CF3CF2CF2CF2-.

[0031] "C" used in this document 3-8 "Cycloalkyl" refers to a saturated monocyclic hydrocarbon ring of 3 to 8 carbon atoms. Examples of such groups include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. If a different number of carbon atoms, e.g., C3-C6, is specified, the definition should be modified accordingly.

[0032] Various embodiments of the present invention are described herein. It will be recognized that features specified in each embodiment may be combined with other specified features to provide additional embodiments. The embodiments listed below represent the present invention:

[0033] Example 1. A compound of Formula I, or a pharmaceutically acceptable salt thereof,

[0034]

[0035] [I]

[0036] In the above formula,

[0037] G 1 and G 2 One of them is selected from linear or branched C1-C5 alkyl; linear or branched C1-C5 alkyloxy; linear or branched C1-C5 haloalkyl; linear or branched C1-C5 haloalkyloxy; halo and 3-7-one-cycloalkyl; and G 1 and G 2 The other one is H; R 1A compound selected from H and linear or branched C1-C5 alkyls, or a pharmaceutically acceptable salt thereof.

[0038] Embodiment 2. In Embodiment 1, G 1 A compound, or a pharmaceutically acceptable salt thereof, selected from linear or branched C1-C5 haloalkyl; linear or branched C1-C5 haloalkyloxy; and 3-7-one cycloalkyl.

[0039] Embodiment 3. In Embodiment 1 or Embodiment 2, G 1 is a compound that is a linear or branched C1-C5 haloalkyl, or a pharmaceutically acceptable salt thereof.

[0040] Embodiment 4. In any one of Embodiments 1 to 3, G 1 A compound that is CF3, or a pharmaceutically acceptable salt thereof.

[0041] Embodiment 5. In Embodiment 1 or Embodiment 2, G 1 is a linear or branched C1-C5 haloalkyloxy, a compound, or a pharmaceutically acceptable salt thereof.

[0042] Embodiment 6. In any one of embodiments 1, 2 and 5, G 1 is a compound, or a pharmaceutically acceptable salt thereof, which is OCF3.

[0043] Embodiment 7. In Embodiment 1 or Embodiment 2, G 1 A compound that is a 3-7-membered cycloalkyl, or a pharmaceutically acceptable salt thereof.

[0044] Embodiment 8. In any one of embodiments 1, 2 and 7, G 1 A compound, or a pharmaceutically acceptable salt thereof, which is cyclopropyl.

[0045] Embodiment 9. In any one of Embodiments 1 to 8, G2 is a compound, or a pharmaceutically acceptable salt thereof, which is H.

[0046] Embodiment 10. In Embodiment 1 or Embodiment 9, G 1 It is a methyl compound, or a pharmaceutically acceptable salt thereof.

[0047] Embodiment 11. In Embodiment 1 or Embodiment 9, G 1 A compound that is OCH3, or a pharmaceutically acceptable salt thereof.

[0048] Embodiment 12. In Embodiment 1 or Embodiment 9, G 1 isopropyl, a compound, or a pharmaceutically acceptable salt thereof.

[0049] Embodiment 13. In Embodiment 1 or Embodiment 9, G 1 Haloins, compounds, or pharmaceutically acceptable salts thereof.

[0050] Embodiment 14. In Embodiment 1, G 1 A compound, or a pharmaceutically acceptable salt thereof, which is H.

[0051] Embodiment 15. In Embodiment 1, G 2 is a methyl, a compound, or a pharmaceutically acceptable salt thereof.

[0052] Example 16. In any one of Examples 1 to 15, a compound of formula (II) or a pharmaceutically acceptable salt thereof:

[0053] [II].

[0054] Example 17. In any one of Examples 1 to 10 and 12 to 13, a compound having the formula (III), or a pharmaceutically acceptable salt thereof:

[0055] [III].

[0056] Example 17a. In any one of Examples 1 to 9, the compound is of Formula (IV):

[0057] [IV],

[0058] In the above equation, G 1 A compound, or a pharmaceutically acceptable salt thereof, selected from linear or branched C1-C5 haloalkyl; linear or branched C1-C5 haloalkyloxy; and 3-7-one cycloalkyl.

[0059] Embodiment 17b. In any one of Embodiments 1 to 9 and 17a, G 1 A compound selected from CF3, OCF3, and cyclopropyl.

[0060] Embodiment 17c. In any one of Embodiments 1 to 9, 17a, and 17b, R 1 A compound selected from H and methyl.

[0061] Example 18. In any one of Examples 1 to 17, a compound selected from the following, or a pharmaceutically acceptable salt thereof:

[0062]

[0063]

[0064]

[0065] .

[0066] Example 18a. A compound selected from the group consisting of the following, or a pharmaceutically acceptable salt thereof, or an optical isomer thereof: N-((4bR,9bR)-1-amino-4b-hydroxy-7-isopropyl-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide; N-(1-amino-4b-hydroxy-10-oxo-7-(trifluoromethyl)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (19); N-((4bR,9bR)-1-amino-4b-hydroxy-10-oxo-7-(trifluoromethyl)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide; N-((4bS,9bS)-1-amino-4b-hydroxy-10-oxo-7-(trifluoromethyl)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide; N-((4bR,9bR)-1-amino-7-bromo-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide; N-(1-amino-7-cyclopropyl-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide; N-((4bR,9bR)-1-amino-7-cyclopropyl-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide; N-((4bS,9bS)-1-amino-7-cyclopropyl-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide; N-(1-amino-4b-hydroxy-10-oxo-7-(trifluoromethoxy)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide; N-((4bR,9bR)-1-amino-4b-hydroxy-10-oxo-7-(trifluoromethoxy)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide;N-((4bS,9bS)-1-amino-4b-hydroxy-10-oxo-7-(trifluoromethoxy)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide; N-(1-amino-7-chloro-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide; N-((4bR,9bR)-1-amino-7-chloro-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide; N-((4bS,9bS)-1-amino-7-chloro-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide; N-(1-amino-4b-hydroxy-7-methyl-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide; N-((4bR,9bR)-1-amino-4b-hydroxy-7-methyl-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide and N-((4bS,9bS)-1-amino-4b-hydroxy-7-methyl-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide; N-(1-amino-4b-hydroxy-8-methyl-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide; N-((4bR,9bR)-1-amino-4b-hydroxy-8-methyl-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide; N-(1-amino-4b-hydroxy-10-oxo-7-(trifluoromethyl)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)-N-methylacetamide; N-((4bR,9bR)-1-amino-4b-hydroxy-7-methoxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide; or pharmaceutically acceptable salts thereof.;

[0067] Embodiment 18b. In Embodiment 1, or Phosphorus compounds, or pharmaceutically acceptable salts thereof.

[0068] Embodiment 19. In any one of embodiments 1 to 18, a compound for the prevention or treatment of a viral disease, a pharmaceutically acceptable salt thereof, or an optical isomer thereof.

[0069] Example 20. A pharmaceutical composition for the prevention or treatment of viral diseases, comprising a compound of any one of Examples 1 to 18, a pharmaceutically acceptable salt or optical isomer thereof, and a pharmaceutically acceptable diluent or excipient.

[0070] Example 21. A combination comprising a compound according to any one of Examples 1 to 18 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to Example 20 and one or more therapeutic active agents.

[0071] Embodiment 22. A method for treating a viral disease comprising the step of administering to a subject a therapeutically effective amount of a compound according to any one of Embodiments 1 to 18 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to Embodiment 20 or a combination according to Embodiment 21.

[0072] Example 23. Use of the compound of Example 19, a pharmaceutically acceptable salt thereof, or an optical isomer thereof, a pharmaceutical composition according to Example 20, or a combination according to Example 21 for the prevention or treatment of viral diseases.

[0073] Embodiment 24. A compound, pharmaceutical composition, method, or use in Embodiment 19, Embodiment 20, Embodiment 21, or Embodiment 23, wherein the viral disease is caused by a coxsackievirus.

[0074] Embodiment 25. A compound, pharmaceutical composition, method, or use in Embodiment 19, Embodiment 20, Embodiment 21, or Embodiment 23, wherein the viral disease is caused by a poliovirus.

[0075] Embodiment 26. In Embodiment 19, Embodiment 20, Embodiment 21 or Embodiment 23, the compound, pharmaceutical composition, method, or use wherein the viral disease is caused by an echovirus.

[0076] Embodiment 27. In Embodiment 19, Embodiment 20, Embodiment 21 or Embodiment 23, the compound, pharmaceutical composition, method, or use wherein the viral disease is caused by an enterovirus.

[0077] Embodiment 28. A compound, pharmaceutical composition, method, or use in Embodiment 19, Embodiment 20, Embodiment 21, or Embodiment 23, wherein the viral disease is caused by a rhinovirus.

[0078] Embodiment 29. A compound, pharmaceutical composition, method, or use in Embodiment 19, Embodiment 20, Embodiment 21, or Embodiment 23, wherein the viral disease is caused by a picornavirus.

[0079] Example 30. In Example 19, Example 20, Example 21, or Example 23, the viral disease is polio, paralysis, acute hemorrhagic conjunctivitis, viral meningitis, hand, foot, and mouth disease, blistering disease, hepatitis A, myositis, myocarditis, pancreatitis, diabetes mellitus, epidemic myalgia, encephalitis, influenza, herpangina, foot-and-mouth disease, asthma, chronic obstructive pulmonary disease, pneumonia, sinusitis, or otitis media, compound, pharmaceutical composition, method, or use.

[0080] Compounds of formulas I, II, or III are novel and are useful as intermediates for the preparation of compounds of formulas (I) to (III) described herein.

[0081] Another embodiment of the present invention provides a compound as described above, or a pharmaceutically acceptable salt thereof, as a pharmaceutical agent.

[0082] The use of a compound of formula I, II, or III, or a pharmaceutically acceptable salt thereof, for the manufacture of a drug for the treatment or prevention of viral diseases and / or infections in humans is also within the scope of the present invention.

[0083] A pharmaceutical composition comprising a compound of formula I, II, or III, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier is included within the scope of the present invention.

[0084] According to a further embodiment of the present invention, the pharmaceutical composition according to the present invention further comprises at least one other antiviral agent in a therapeutically effective amount.

[0085] The present invention also provides the use of a pharmaceutical composition as described above for the treatment of a viral infection in humans who have or are at risk of having an infection.

[0086] The present invention also provides the use of a pharmaceutical composition as described above for the treatment of viral diseases in humans who have or are at risk of having the disease.

[0087] Another aspect of the present invention comprises a method for treating or preventing viral diseases and / or infections in humans, comprising the step of administering to a human an antiviral effective amount of a compound of the present invention, a pharmaceutically acceptable salt thereof, or a composition as described above, in combination with at least one other antiviral agent administered alone, together, or separately.

[0088] A further aspect of the present invention relates to an article of manufacture comprising a composition effective for treating viral diseases and / or infections; and a packaging material comprising a label indicating that said composition may be used to treat diseases and / or infections caused by viruses; wherein said composition comprises a compound of Formula I, II, or III according to the present invention or a pharmaceutically acceptable salt thereof.

[0089] Another aspect of the present invention relates to a method for inhibiting the replication of a virus, comprising the step of exposing a virus to an effective amount of a compound of formula I, II, or III, or a salt thereof, under conditions in which the replication of the virus is inhibited. This method may be carried out in vitro or in vivo.

[0090] The use of a compound of formula I, II, or III, or a salt thereof, for inhibiting viral replication is further included within the scope of the present invention.

[0091] In one embodiment, the present invention provides a pharmaceutical composition comprising a compound of the present invention and another therapeutic agent(s). Optionally, the pharmaceutical composition may comprise a pharmaceutically acceptable carrier as described above. In some embodiments, the compound of Formula I, II, or III is co-administered with at least one additional agent selected from a viral inhibitor or a vaccine.

[0092] These additional formulations may be combined with the compound of the present invention to form a single pharmaceutical formulation. Alternatively, these additional formulations may be administered separately to a patient as part of a multi-dose formulation, for example, using a kit. These additional formulations may be administered to the patient before, simultaneously with, or after the administration of the compound of the present invention or its pharmaceutically acceptable salt.

[0093] The dosage range of the compound of the present invention applicable per day is generally 0.01 to 100 mg / (kg body weight), and occasionally 0.1 to 50 mg / (kg body weight). Each dosage unit may conveniently contain 5% to 95% of the active compound (w / w). Occasionally, such preparations contain 20% to 80% of the active compound.

[0094] The actual pharmaceutically effective or therapeutic dose will vary depending on factors known to those skilled in the art, such as the patient's age and weight, route of administration, and severity of the disease. In any case, the combination will be administered at a dose and by a method that allows the pharmaceutically effective dose to be delivered based on the patient's unique condition.

[0095] Where the composition of the present invention comprises a combination of the compound of the present invention and one or more additional therapeutic or prophylactic agents, said compound and additional agents may be present at a dosage level of about 10 to 100%, for example, about 10 to 80% of the dosage typically administered in monotherapy.

[0096] Antiviral agents considered for use in such combination therapy include agents (compounds or biological agents) effective in inhibiting viral production and / or replication in humans, which include but are not limited to agents that interfere with host or viral mechanisms necessary for viral production and / or replication in humans.

[0097] Many of the compounds of the present invention contain one or more chiral centers. These compounds may be prepared and used as single isomers or mixtures of isomers. Methods for separating isomers, including diastereomers and enantiomers, are known in the art, and examples of suitable methods are described herein. In certain embodiments, the compounds of the present invention are used as substantially pure single isomers, which means that at least 90% of the compound sample is a specific isomer and less than 10% of the sample is any other isomer or a mixture of isomers. In some embodiments, at least 95% of the sample is a single isomer. When the difference in in vitro activity between isomers is relatively small, for example, less than about 4 times, the single isomer may be selected based on the level of activity against viral replication in cell culture using methods such as those described herein: the isomer having a lower IC-50 or EC-50 may be selected.

[0098] The compounds of the present invention can be synthesized by the general synthesis routes below, and specific examples thereof are described in more detail in the examples.

[0099] The present invention also provides a method for preparing a compound of formula I, II, or III as described herein and an intermediate useful for the preparation of a compound of formula I, II, or III.

[0100] The present invention further comprises any variation of the method wherein an intermediate product obtainable at any step is used as a starting material and the remaining steps are performed, the starting material is formed in situ under reaction conditions, or the reaction component is used in the form of its salt or an optically pure substance.

[0101] The present invention also relates to a method of such form in which a compound obtainable as an intermediate at any step of the method is used as a starting material and the remaining method steps are performed, or the starting material is formed under reaction conditions or used in the form of a derivative, e.g., a protected form or a salt, or a compound obtainable by the method according to the present invention is generated under method conditions and further processed in situ.

[0102] The terms "optical isomer" or "stereoisomer" refer to any various stereoisomer configurations that may exist for a given compound of the present invention, including geometric isomers. Substituents are understood to be those that can be attached to the chiral center of a carbon atom. The term "chiral" refers to a molecule having the characteristic that it cannot be superimposed on a mirror image partner, whereas the term "achiral" refers to a molecule that can be superimposed on a mirror image partner. Accordingly, the present invention includes enantiomers, diastereomers, or racemics of a compound. "Enantiomers" are a pair of stereoisomers that are mirror images of each other and cannot be superimposed. A 1:1 mixture of a pair of enantiomers is a "racemic" mixture. The above term is used to refer to a racemic mixture as necessary. "Diastereomers" are stereoisomers that have at least two asymmetric atoms but are not mirror images of each other. Absolute stereochemistry is designated according to the Cahn-Ingold-Prelog RS system. If the compound is a pure enantiomer, the stereochemistry at each chiral carbon may be designated as R or S. Split compounds with unknown absolute configuration may be designated as (+) or (-) depending on the direction of rotation of plane polarization at the wavelength of the sodium D line (left-handed or right-handed). Since the specific compounds described herein contain one or more asymmetric centers or axes, they may form enantiomers, diastereomers, and other stereoisomers that can be defined as (R)- or (S)- in terms of absolute stereochemistry.

[0103] Depending on the choice of starting materials and procedures, the compound may exist in the form of one of the possible isomers or as a mixture thereof, for example as a pure optical isomer, or as a mixture of isomers, such as a racemic mixture and a diastereomer mixture, depending on the number of asymmetric carbon atoms. The present invention implies the inclusion of all such possible stereoisomers, including racemic mixtures, diastereomer mixtures, and optically pure forms. Optically active (R)- and (S)-isomers may be prepared using chiral synthons or chiral reagents, or may be cleaved using conventional techniques. If the compound contains a double bond, the substituent may be in an E or Z configuration. If the compound contains a disubstituted cycloalkyl, the cycloalkyl substituent may have a cis or trans configuration. All tautomers are also intended to be included.

[0104] Any obtained mixture of isomers can be separated into pure or substantially pure geometric or optical isomers or diastereomers based on physicochemical differences of the components, for example, by chromatography and / or fractional crystallization.

[0105] Any obtained racemic product or intermediate may be divided into optical antipodes by known methods, for example, by separating the diastereomer salts thereof obtained with an optically active acid or base and liberating an optically active acidic or basic compound. In particular, the basic portion may be used to divide the compound of the present invention into its optical antipodes by fractional crystallization of a salt formed with an optically active acid, for example, tartaric acid, dibenzoyl tartaric acid, diacetyl tartaric acid, di-O,O'-p-toluoyly tartaric acid, mandelic acid, malic acid, or camphor-10-sulfonic acid. The racemic product may also be divided by chiral chromatography, for example, high-pressure liquid chromatography (HPLC), using a chiral adsorbent.

[0106] Additionally, the compounds of the present invention (including their salts) may also be obtained in the form of their hydrates or may comprise other solvents used for their crystallization. The compounds of the present invention may, by nature or intentionally, form solvates with pharmaceutically acceptable solvents (including water); thus, the present invention is intended to include both solvated and non-solvent forms. The term "solvate" refers to a molecular complex of the compounds of the present invention (including their pharmaceutically acceptable salts) and one or more solvent molecules. Such solvent molecules are those commonly used in the pharmaceutical field and known to be harmless to recipients, such as, for example, water, ethanol, etc. The term "hydrate" refers to a complex in which the solvent molecule is water.

[0107] The compounds of the present invention (including their salts, hydrates, and solvates) may form polymorphs, either essentially or intentionally.

[0108] As used herein, the terms “salt” or “salts” refer to acid-addition salts or base-addition salts of the compounds of the present invention. “Salt” specifically includes “pharmaceutically acceptable salts.” The term “pharmaceutically acceptable salt” refers to a salt that retains the biological effects and properties of the compounds of the present invention and is typically not biologically or otherwise undesirable. In many cases, the compounds of the present invention may form acid and / or base salts due to the presence of amino groups and / or carboxyl groups or similar groups.

[0109] Pharmaceutically acceptable acid addition salts may be formed of inorganic and organic acids, for example, acetate, aspartate, benzoate, besylate, bromide / hydrobromide, bicarbonate / carbonate, bisulfate / sulfate, campersulfonate, chloride / hydrochloride, chlortheophilonate, citrate, ethanedisulfonate, fumarate, gluteptate, gluconate, glucuronate, hippurate, hydroiodide / iodide, isethionate, lactate, lactobionate, lauryl sulfate, maleate, maleate, malonate, mandelate, mesylate, methyl sulfate, naphthoate, lead sylate, nicotinate, nitrate, octadecanoate, oleate, oxalate, palmitate, There are pamoate, phosphate / hydrogen phosphate / dihydrogen phosphate, polygalacturonate, propionate, stearate, succinate, sulfosalicylate, tartrate, tosylate, and trifluoroacetate salts.

[0110] Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc.

[0111] Organic acids from which salts may be derived include, for example, acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, sulfosalicylic acid, etc. Pharmaceutically acceptable base addition salts may be formed from inorganic and organic bases.

[0112] Inorganic bases from which salts can be derived include, for example, ammonium salts and metals of groups I to XII of the periodic table. In a given embodiment, the salt is derived from sodium, potassium, ammonium, calcium, magnesium, iron, silver, zinc, and copper; particularly suitable salts include ammonium, potassium, sodium, calcium, and magnesium salts.

[0113] Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines including naturally derived substituted amines, cyclic amines, basic ion exchange resins, etc. Specific organic amines include isopropylamine, benzathine, cholinenate, diethanolamine, diethylamine, lysine, meglumine, piperazine, and tromethamine.

[0114] Pharmaceutically acceptable salts of the present invention may be synthesized from basic or acidic portions by conventional chemical methods. Generally, such salts may be prepared by reacting the free acid form of these compounds with a stoichiometric amount of a suitable base (e.g., Na, Ca, Mg, or K hydroxide, carbonate, bicarbonate, etc.) or by reacting the free base form of these compounds with a stoichiometric amount of a suitable acid. Such reactions are typically carried out in water or an organic solvent, or a mixture of both. Generally, where feasible, the use of a non-aqueous medium such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile is preferred. A list of additional suitable salts is provided, for example, in the literature ["Remington's Pharmaceutical Sciences", 20th ed., Mack Publishing Company, Easton, Pa., (1985)]; and can be found in the literature ["Handbook of Pharmaceutical Salts: Properties, Selection, and Use" by Stahl and Wermuth (Wiley-VCH, Weinheim, Germany, 2002)].

[0115] Any of the specified chemical formulas in this specification is a non-natural isotope distribution, e.g., deuterium or 13 C or 15 It is intended to represent the isotope-labeled form as well as the unlabeled form of the compound of the present invention having up to three atoms having N-rich sites. The isotope-labeled compound has a structure represented by the formulas specified herein, except that one or more atoms are substituted with atoms having selected atomic masses or mass numbers other than their natural abundance mass distributions. Examples of isotopes that may be usefully over-incorporated into the compound of the present invention are, respectively, isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, e.g. 2 H,3 H, 11 C, 13 C, 14 C, 15 N, 18 F, 31 P, 32 P, 35 S, 36 Cl, 125 The present invention includes I. The present invention relates to various isotope-labeled compounds of the present invention, for example, 3 H and 14 Radioactive isotopes such as C, or 2 H and 13 It includes compounds in which non-radioactive isotopes, such as C, are present at substantially higher levels than the normal isotopic distribution. These isotope-labeled compounds are used in metabolic studies, including drug or substrate tissue distribution assays (e.g., 14 Using C), reaction rate studies (e.g. 2 H or 3 It is useful for detection or imaging techniques, such as positron emission tomography (PET) or single-photon emission computed tomography (SPECT), or for radiation therapy of patients. In particular, the present invention 18 F-labeled compounds may be particularly desirable for PET or SPECT studies. The isotope-labeled compounds of the present invention can generally be prepared by ordinary techniques known to those skilled in the art using a suitable isotope-labeled reagent instead of the unlabeled reagents commonly used, or by methods similar to those described in the attached examples and preparation examples. Labeled samples may be useful for significantly low isotope incorporation, such as when radiolabeling is used to detect trace amounts of compounds.

[0116] Also, heavier isotopes, especially deuterium (i.e., 2More extensive substitution with H or D may provide specific therapeutic benefits due to greater metabolic stability, such as increased in vivo half-life, reduced required dose, or improved therapeutic index. In this regard, deuterium is considered a substituent of the compounds of the present invention, and typically, a sample of a compound having deuterium as a substituent is understood to have at least 50% deuterium incorporation at the labeled position(s). The concentration of these heavier isotopes, specifically deuterium, may be defined by the isotope enrichment factor. As used herein, the term "isotope enrichment factor" refers to the ratio between the isotopic abundance of a specified isotope and its natural abundance. When a substituent in a compound of the present invention is a designated deuterium, such compound has an isotope enrichment factor of at least 3500 (52.5% deuterium incorporation in each designated deuterium atom), at least 4000 (60% deuterium incorporation), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), at least 6600 (99% deuterium incorporation), or at least 6633.3 (99.5% deuterium incorporation) for each designated deuterium atom.

[0117] The pharmaceutically acceptable solvate according to the present invention is one in which the crystallization solvent can be uniatomicly substituted, e.g., D2O, d 6 -Acetone, d 6 - Contains DMSO.

[0118] Compounds of the present invention comprising groups capable of acting as donors and / or acceptors for hydrogen bonding can form cocrystals with a suitable cocrystal-forming agent. Such cocrystals can be prepared from compounds of the present invention by known cocrystal-forming procedures. Such procedures include contacting compounds of the present invention with a cocrystal-forming agent in a solution under conditions of grinding, heating, co-sublimation, co-melting, or crystallization, and isolating the cocrystals formed thereby. Suitable cocrystal-forming agents include those described in International Publication WO 2004 / 078163. Accordingly, the present invention further provides cocrystals comprising compounds of the present invention.

[0119] All methods described herein may be performed in any suitable order unless otherwise indicated herein or clearly contradictory in the context. The use of any examples or exemplary language (e.g., “e.g., for example”) provided herein is merely for better illustrating the invention and does not limit the scope of the invention as otherwise claimed.

[0120] The compounds of the present invention may be administered by known methods including oral, parenteral, and inhalation. In certain embodiments, the compounds of the present invention are administered orally as pills, lozenges, troches, capsules, solutions, or suspensions. In other embodiments, the compounds of the present invention are administered by injection or infusion. Infusion is typically performed intravenously over a period of about 15 minutes to 4 hours. In other embodiments, the compounds of the present invention are administered intranasally or by inhalation; the inhalation method is particularly useful for the treatment of respiratory infections. The compounds of the present invention exhibit oral bioavailability, and thus, in some embodiments, the compounds may be administered orally.

[0121] The compound of the present invention may also be used in combination with other agents (combination partners), for example, additional antiviral agents of formula I or not of formula I, to treat viral infections in subjects.

[0122] The term “combination” means a fixed combination of a single unit formulation as individual formulations suitable for use together simultaneously or sequentially, or a kit of parts for co-administration in which the compound of the present invention and the co-administration partner may be administered independently, simultaneously, or, in particular, within a time interval in which the co-administration partners cooperate, e.g., to produce a synergistic effect, or any combination thereof.

[0123] In a given embodiment of the present invention, the compound of the present invention is used in combination with a second antiviral agent, such as one named herein.

[0124] A second antiviral agent may be administered in combination with a compound of the present invention, wherein the second antiviral agent is administered before, simultaneously with, or after the administration of the compound or compounds of the present invention. If simultaneous administration of the second agent and the compound of the present invention is required and the route of administration is the same, the compound of the present invention may be formulated in the same formulation together with the second agent. Examples of formulations containing the compound and the second agent of the present invention are tablets or capsules.

[0125] In some embodiments, the combination of the compound of the present invention and the second antiviral agent may provide synergistic activity. The compound of the present invention and the second antiviral agent may be administered together, separately, but simultaneously or sequentially.

[0126] The "effective amount" of a compound is an amount necessary or sufficient to treat or prevent viral infections and / or diseases or conditions described herein. In one example, the effective amount of a viral inhibitor of Formula I is an amount sufficient to treat a viral infection in a subject. The effective amount may vary depending on factors such as the size and weight of the subject, the type of disease, or specific compounds of the present invention. For example, the selection of compounds of the present invention may influence the composition of the "effective amount." A person skilled in the art will be able to determine the effective amount of the compounds of the present invention without excessive experimentation by studying the factors included herein.

[0127] The administration schedule may affect the composition of the effective dose. The compounds of the present invention may be administered to a subject before or after the onset of a viral infection. Additionally, multiple divided doses and timed doses may be administered daily or sequentially, or the dose may be continuously infused or as a bolus injection. Furthermore, the dosage of the compound(s) of the present invention may be proportionally increased or decreased as an urgent therapeutic or prophylactic situation arises.

[0128] The compounds of the present invention may be used to treat conditions, disorders, or diseases as described herein, or to prepare pharmaceutical compositions for use in treating these diseases. The present invention provides a method for using the compounds of the present invention in the treatment of these diseases or a method for preparing pharmaceutical compositions containing the compounds of the present invention for the treatment of these diseases.

[0129] The term “pharmaceutical composition” includes preparations suitable for administration to mammals, e.g., humans. When a compound of the present invention is administered to a mammal, e.g., humans as a medicine, it may be provided as a pharmaceutical composition containing, either by itself or, e.g., one pharmaceutically acceptable carrier or optionally in combination with two or more pharmaceutically acceptable carriers, at least one compound of formula (I) or any subgenus thereof as an active ingredient in an amount of 0.1 to 99.5% (sometimes 0.5 to 90%).

[0130] The phrase “pharmaceuticalally acceptable carrier” includes pharmaceutically acceptable substances, compositions, or vehicles recognized in the art and suitable for administering the compounds of the present invention to mammals. Carriers include liquid or solid fillers, diluents, excipients, solvents, or encapsulating agents involved in transporting or carrying the target formulation from one organ or part of the body to another organ or part of the body. Each carrier must be “acceptable” in the sense that it is compatible with other components of the formulation and is not harmful to the patient. Some examples of substances that may serve as pharmaceutically acceptable carriers include sugars, e.g., lactose, glucose, and sucrose; starches, e.g., corn starch and potato starch; cellulose and derivatives thereof, e.g., sodium carboxymethylcellulose, ethyl cellulose, and cellulose acetate; tragacanth powder; malt; gelatin; talc; excipients, e.g., cocoa butter and suppository waxes; oils, e.g., peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; Glycols, e.g., propylene glycol; polyols, e.g., glycerin, sorbitol, mannitol, and polyethylene glycol; esters, e.g., ethyl oleate and ethyl laurate; agar; buffers, e.g., magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffer; and other non-toxic, compatible materials used in pharmaceutical formulations. Typically, pharmaceutically acceptable carriers are sterile and / or substantially pyrogen-free.

[0131] In addition to wetting agents, emulsifiers, and lubricants, such as sodium lauryl sulfate and magnesium stearate, coloring agents, release agents, coating agents, sweeteners, flavoring agents and fragrances, preservatives, and antioxidants may also be present in the composition.

[0132] Examples of pharmaceutically acceptable antioxidants include water-soluble antioxidants, such as ascorbic acid, cysteine ​​hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, etc.; fat-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, α-tocopherol, etc.; and metal chelating agents, such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, etc.

[0133] The formulations of the present invention include those suitable for oral, nasal, inhalation, topical, transdermal, buccal, sublingual, rectal, vaginal, and / or parenteral administration. The formulations may conveniently be provided in a single-dose form and may be prepared by any method well known in the pharmaceutical field. The amount of the active ingredient that may be combined with a carrier material to produce a single-dose form will generally be the amount of the compound that yields a therapeutic effect. Generally, out of 100%, this amount will be in the range of about 1% to about 99% of the active ingredient, occasionally about 5% to about 70%, and occasionally about 10% to about 30%.

[0134] A method for preparing such a formulation or composition comprises the step of associating a compound of the present invention with a carrier and optionally one or more auxiliary components. Generally, the formulation is prepared by uniformly and closely bonding the compound of the present invention with a liquid carrier, a fine solid carrier, or both, and then forming a product as needed.

[0135] Formulations of the present invention suitable for oral administration may be in the form of capsules, cachets, pills, tablets, lozenges (flavored base, e.g., generally using sucrose and acacia or tragacanth), powders, granules, or as solutions or suspensions in aqueous or non-aqueous liquids, or as oil-in-water or water-in-oil liquid emulsions, or as elixirs or syrups, or as flavorings (pastilles, inert bases, e.g. using gelatin and glycerin, or sucrose and acacia), and / or mouthwashes, each containing a predetermined amount of the compound of the present invention as an active ingredient. The compound of the present invention may also be administered as a bolus, an electric, or a paste.

[0136] In the solid formulation of the present invention for oral administration (capsules, tablets, pills, coated tablets, powders, granules, etc.), the active ingredient is mixed with one or more pharmaceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and / or any one of the following: fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol and / or silicic acid; binders, such as carboxymethylcellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose and / or acacia; humectants, such as glycerol; disintegrants, such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate; solution retarding agents, such as paraffin; absorption promoters, such as quaternary ammonium compounds; wetting agents, such as cetyl alcohol and glycerol monostearate; absorbents, such as kaolin and bentonite clay; Lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof; and coloring agents. In the case of capsules, tablets, and pills, the pharmaceutical composition may also include a buffer. A similar type of solid composition may also be used as a filler for soft and hard gelatin capsules using excipients such as high molecular weight polyethylene glycol, as well as lactose or lactose.

[0137] Tablets may be manufactured by compression or molding, optionally using one or more auxiliary components. Compressed tablets may be manufactured using a binder (e.g., gelatin or hydroxypropylmethylcellulose), a lubricant, an inert diluent, a preservative, a disintegrant (e.g., sodium starch glycolate or cross-linked sodium carboxymethylcellulose), a surfactant, or a dispersant. Molded tablets may be manufactured by molding a mixture of powdered compounds soaked in an inert liquid diluent using a suitable machine.

[0138] Tablets and other solid formulations of the pharmaceutical composition of the present invention, such as coated tablets, capsules, pills, and granules, may optionally be scored or manufactured with coatings and shells, such as enteric coatings and other coatings well known in the field of pharmaceutical formulation. They may also be formulated to provide sustained or controlled release of the active ingredient therein, for example, by using hydroxypropylmethylcellulose in various proportions to provide a desired release profile, different polymer matrices, liposomes, and / or microspheres. They may be sterilized, for example, by filtering through a bacterial retention filter or by incorporating a sterile agent in the form of a sterile solid composition that can be dissolved in sterile water or some other sterile injectable medium immediately before use. Such compositions may also optionally contain an opacifier and may be compositions that release the active ingredient(s) alone or, for example, in a delayed manner at specific sites of the gastrointestinal tract. Examples of embedding compositions that may be used include polymeric materials and waxes. The active ingredient may also exist in a microencapsulated form with one or more of the aforementioned excipients, where appropriate.

[0139] Liquid formulations for oral administration of the compounds of the present invention include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active ingredient, the liquid formulations may contain, in addition to the active ingredient, inert diluents commonly used in the art, such as water or other solvents, solubilizers and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (particularly cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofuryl alcohol, fatty acid esters of polyethylene glycol and sorbitan, and mixtures thereof.

[0140] In addition to inert diluents, oral compositions may also include adjuvants, such as humectants, emulsifying and suspending agents, sweeteners, flavoring agents, coloring agents, fragrances, and preservatives.

[0141] In addition to the active compound, the suspension may contain suspending agents such as, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar and tragacanth, and mixtures thereof.

[0142] A formulation of the pharmaceutical composition of the present invention for rectal or vaginal administration may be provided as a suppository, which may be prepared by mixing one or more compounds of the present invention with one or more suitable non-irritating excipients or carriers, such as cocoa butter, polyethylene glycol, suppository wax, or salicylate, which is solid at room temperature but liquid at body temperature and will melt in the rectal or vaginal cavity to release the active compound.

[0143] The formulations of the present invention suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, foams, or spray formulations containing such carriers known to be suitable in the art.

[0144] Formulations for topical or transdermal administration of the compounds of the present invention include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, and inhalants. The active compound may be mixed with a pharmaceutically acceptable carrier under sterile conditions, and, if necessary, with any preservative, buffer, or propellant.

[0145] In addition to the active compound of the present invention, ointments, pastes, creams, and gels may contain excipients such as animal and vegetable fats, oils, waxes, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc, and zinc oxide, or mixtures thereof.

[0146] In addition to the compounds of the present invention, the powder and spray may contain excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicate, and polyamide powder, or mixtures of these materials. The spray may further contain conventional propellants, such as chlorofluorohydrocarbons, and volatile unsubstituted hydrocarbons, such as butane and propane.

[0147] Transdermal patches have the additional advantage of providing controlled delivery of the compounds of the present invention to the body. Such formulations can be prepared by dissolving or dispersing the compounds in a suitable medium. Absorption promoters may also be used to increase the flux of the compounds through the skin. This flux rate can be controlled by providing a rate-controlling membrane or by dispersing the active compounds in a polymer matrix or gel.

[0148] Ophthalmic formulations, ophthalmic ointments, powders, solutions, etc. are also considered to be within the scope of the present invention.

[0149] A pharmaceutical composition of the present invention suitable for parenteral administration comprises one or more compounds of the present invention in combination with one or more pharmaceutically acceptable carriers, such as sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions, or emulsions, or in combination with sterile powders that can be reconstituted in a sterile injectable or dispersion immediately before use, and may contain an antioxidant, a buffer, a bacteriostatic agent, a solute that makes the formulation isotonic with the blood of the intended recipient, or a suspending agent or a thickening agent.

[0150] Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical composition of the present invention include water, ethanol, glycol ethers, polyols (e.g., glycerol, propylene glycol, polyethylene glycol, etc.), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate. Suitable fluidity can be maintained, for example, by the use of a coating material, such as lecithin, by maintaining the required particle size in the case of a dispersion, and by the use of a surfactant.

[0151] These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifiers, and dispersants. Prevention of microbial action can be ensured by including various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol sorbic acid, etc. Additionally, it may be desirable to include isotonic agents, such as sugars and sodium chloride, in the composition. Furthermore, sustained absorption of the injectable formulation may occur through the incorporation of agents that delay absorption, such as aluminum monostearate and gelatin.

[0152] In some cases, to prolong the effect of the drug, it is desirable to delay the absorption of the drug via subcutaneous or intramuscular injection. This can be achieved by using a liquid suspension of a crystalline or amorphous material that is poorly soluble. In this case, the rate of drug absorption depends on its dissolution rate, which can ultimately vary depending on crystal size and crystal form. Alternatively, delayed absorption of parenterally administered drug forms is achieved by dissolving or suspending the drug in an oil vehicle.

[0153] Injectable depot formulations are prepared by forming a microcapsule matrix of the target compound in biodegradable polymers such as polylactide-polyglycolide. Depending on the drug-to-polymer ratio and the characteristics of the specific polymer used, the drug release rate can be controlled. Other examples of biodegradable polymers include poly(orthoesters) and poly(anhydrides). Additionally, depot injectable formulations are prepared by encapsulating the drug in liposomes or microemulsions suitable for body tissues.

[0154] The formulations of the present invention may be administered orally, parenterally, topically, or rectally. Of course, they are administered in a form suitable for each route of administration. For example, they are administered in the form of tablets or capsules by injection or inhalation; as eye drops, ointments, suppositories, etc. by injection, infusion, or inhalation; topically by lotion or ointment; and rectally by suppository.

[0155] As used herein, the terms “parenteral administration” and “parenterally administered” generally refer to methods of administration other than by injection, enteral administration, and local administration, and include, but are not limited to, intravenous, intramuscular, intra-arterial, intradural, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtubercular, subcutaneous, subepidermal, intra-articular, subcapsular, subarachnoid, intraspinal, and intrasternal injections and infusions. Intravenous infusion is occasionally a method of delivering the compounds of the present invention. Infusions may be used to deliver once-daily doses or multiple doses. In some embodiments, the compounds of the present invention are administered by infusion over intervals of 15 minutes to 4 hours, typically 0.5 to 3 hours. Such infusions may be used once a day, twice a day, or up to three times a day.

[0156] As used herein, the terms “systemic administration,” “systemically administered,” “peripheral administration,” and “peripherally administered” refer to the administration of compounds, drugs, or other substances—excluding direct administration to the central nervous system—that enter the patient’s body tissues and apply to metabolism and other similar processes, e.g., subcutaneous administration.

[0157] These compounds may be administered to humans and other animals for therapy orally, intranasally, e.g. as a spray, rectally, vaginally, parenterally, intracervix, and topically, including buccally and sublingually, as a powder, ointment, or drops.

[0158] Regardless of the selected route of administration, the compounds of the present invention and / or the pharmaceutical compositions of the present invention that can be used in the form of appropriate hydrates are formulated into pharmaceutically acceptable formulations by conventional methods known to those skilled in the art.

[0159] The actual dosage level of the active ingredient in the pharmaceutical composition of the present invention can be varied to obtain an amount of the active ingredient effective in achieving a desired therapeutic response for a specific patient, composition, and method of administration, without being toxic to the patient.

[0160] The selected dosage level will depend on various factors, including the activity of the specific compound of the present invention used or its ester, salt, or amide, the route of administration, the timing of administration, the excretion rate of the specific compound used, the duration of treatment, other drugs, compounds, and / or substances used in combination with the specific compound used, the age, gender, weight, condition, overall health status, and previous medical history of the patient to be treated, and similar factors well known in the medical field.

[0161] A physician or veterinarian with ordinary skill in the art can easily determine and prescribe the effective amount of the necessary pharmaceutical composition. For example, a physician or veterinarian may start the dosage of the compound of the present invention used in the pharmaceutical composition at a level lower than the amount required to achieve the desired therapeutic effect, and gradually increase the dosage until the desired effect is achieved.

[0162] Generally, an appropriate daily dose of the compound of the present invention will be the amount of the compound that is the lowest effective dose to produce a therapeutic effect. This effective dose will generally vary depending on the factors mentioned above. Typically, when used for the indicated effect, the intravenous and subcutaneous doses of the compound of the present invention to a patient will be in the range of about 0.0001 to about 100 mg / (kg·day body weight), occasionally about 0.01 to about 50 mg / (kg·day body weight), and occasionally about 0.1 to about 20 mg / (kg·day body weight). The effective dose is an amount that prevents or treats a viral infection.

[0163] If necessary, the daily effective amount of the active compound may be administered as a once-daily dose, or optionally as unit formulations, as 2, 3, 4, 5, 6, or more sub-dose doses administered individually at appropriate intervals throughout the day. Compounds delivered orally or by inhalation are generally administered 1 to 4 times per day. Compounds delivered by injection are generally administered once per day or every other day. Compounds delivered by infusion are generally administered 1 to 3 times per day. When multiple doses are administered within a day, the doses may be administered at intervals of about 4 hours, about 6 hours, about 8 hours, or about 12 hours.

[0164] While it is possible for the compound of the present invention to be administered alone, it is also possible to administer the compound as a pharmaceutical composition as described herein. Accordingly, a method of using the compound of the present invention comprises administering the compound as a pharmaceutical composition, wherein at least one compound of the present invention is mixed with a pharmaceutically acceptable carrier prior to administration.

[0165] General synthesis procedure

[0166] Compounds as described herein can be synthesized by the following general synthesis routes, and specific examples thereof are described in more detail in the examples.

[0167] All starting materials, building blocks, reagents, acids, bases, dehydrating agents, solvents, and catalysts used to synthesize the compounds of the present invention may be commercially available or prepared by organic synthesis methods known to those skilled in the art (Reference [Houben-Weyl 4th Ed. 1952, Methods of Organic Synthesis, Thieme, Volume 21]).

[0168] Abbreviation list

[0169]

[0170]

[0171] The compounds of the present invention are prepared from universally available compounds using procedures known to those skilled in the art in terms of the examples and reaction schemes provided herein.

[0172] Within the scope of this specification, easily removable deceptions that are not components of a specific desired end product of the compound of the present invention are referred to as “protective groups” unless the context otherwise indicates. The protection of functional groups by such protecting groups, the protecting groups themselves, and their cleavage reactions are, for example, in standard reference studies, such as [Science of Synthesis: Houben-Weyl Methods of Molecular Transformation. Georg Thieme Verlag, Stuttgart, Germany. 2005. 41627 pp. (URL: http: / / www.science-of-synthesis.com (Electronic Version, 48 Volumes)]; [JFW McOmie, “Protective Groups in Organic Chemistry”, Plenum Press, London and New York 1973], [TW Greene and PGM Wuts, “Protective Groups in Organic Synthesis”, Third edition, Wiley, New York 1999], [“The Peptides”; Volume 3 (editors: E. Gross and J. Meienhofer), Academic Press, London and New York 1981], "Methoden der Organischen Chemie" (Methods of Organic Chemistry), Houben Weyl, 4th edition, Volume 15 / I, Georg Thieme Verlag, Stuttgart 1974], H.-D. Jakubke and H.This is described in Jeschkeit, "Aminosauren, Peptide, Proteine" (Amino acids, Pep-tides, Proteins), Verlag Chemie, Weinheim, Deerfield Beach, and Basel 1982), and in the literature [Jochen Lehmann, "Chemie der Kohlenhydrate: Monosaccharides und Derivate" (Chemistry of Carbohydrates: Monosaccharides and Derivatives), Georg Thieme Verlag, Stuttgart 1974]. The characteristic of the protecting group is that the protecting group can be easily removed (i.e., without the occurrence of unwanted secondary reactions) by, for example, solvent digestion, reduction, photolysis, or alternatively under physiological conditions (e.g., enzymatic cleavage).

[0173] Salts of the compounds of the present invention having at least one salt-forming group can be prepared by methods known in themselves. For example, salts of the compounds of the present invention having an acidic group can be formed, for example, by treating the compound with an alkali metal salt of a suitable organic carboxylic acid, a metal compound such as the sodium salt of 2-ethylhexanoic acid, an organic alkali metal or alkaline earth metal compound, such as a corresponding hydroxide, carbonate or hydrogen carbonate, such as sodium hydroxide or potassium hydroxide, carbonate or hydrogen carbonate, a corresponding calcium compound, or ammonia or a suitable organic amine, wherein stoichiometric amounts or only small excess amounts of the salt-forming agent are occasionally used. Acid addition salts of the compounds of the present invention are obtained by conventional methods, for example, by treating the compound with an acid or a suitable anion exchange reagent. The internal salt of the compound of the present invention, which includes acidic salt and basic salt forming groups, such as free carboxyl groups and free amino groups, can be formed, for example, by neutralizing a salt such as an acid addition salt to an isoelectric point using a weak base or by treating it with an ion exchanger.

[0174] Salts can be converted into free compounds by conventional methods; metal and ammonium salts can be converted, for example, by treatment with a suitable acid, and acid addition salts can be converted, for example, by treatment with a suitable basic agent.

[0175] A mixture of isomers obtainable according to the present invention can be separated into individual isomers by a known method; diastereomers can be separated, for example, by partitioning between multiphase solvent mixtures, recrystallization and / or, for example, by chromatographic separation on silica gel, or, for example, by medium-pressure liquid chromatography on a reverse-phase column, and racemic mixtures can be separated, for example, by forming a salt with an optically pure salt-forming reagent and, for example, by fractional crystallization or by chromatography on an optically active column material.

[0176] Intermediates and final products can be worked up and / or purified according to standard methods using, for example, chromatographic methods, partitioning methods, (re)crystallization, etc.

[0177] Examples

[0178] The present invention is further illustrated by the following examples, which should not be construed as limiting. The analysis used throughout the examples is well established in the art: the demonstration of efficacy in such analysis is generally considered to predict efficacy in the subject.

[0179] The compounds of the present invention can be prepared by organic synthesis methods known to those skilled in the art with reference to the following reaction schemes and examples. A general method for synthesizing the compound of formula (I) is provided in the reaction scheme below.

[0180] High-resolution mass spectroscopy by LC-MS

[0181] ESI-MS data were recorded using an LTQ-XL Orbitrap mass spectrometer (ThermoFisher Scientific) with an electrospray ionization source. The resolution of the MS system was approximately 30,000. Drug candidates were injected into the mass spectrometer from the sample probe via a UPLC (Acquity, Waters). Separation was performed on an Acquity UPLC BEH C18 1×50 mm column at a flow rate of 0.15 mL / min with a gradient from 5% to 95% within 3 minutes. Solvent A was water containing 0.1% trifluoroacetic acid, and Solvent B was 75% methanol and 25% isopropyl alcohol containing 0.1% trifluoroacetic acid. The mass accuracy of the system was found to be <5 ppm.

[0182] Examples 1 and 2: N-((4bR,9bR)-1-amino-4b-hydroxy-7-isopropyl-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide

[0183]

[0184] Reaction Equation 1:

[0185]

[0186] 4-nitroisobenzofuran-1,3-dione (2):

[0187] An initial suspension of 1 (1.0 kg, 4.7 mol) of 3-nitrophthalic acid in 1 Ltr of Ac2O was refluxed at 140°C for 2.5 hours. Afterward, it was cooled to 80°C and slowly added to 4 Ltr of diethyl ether while vigorously stirring. The precipitate was collected by filtration through a Buckner funnel and washed with Et2O to obtain the product as a solid.

[0188] Ethyl 4-nitro-1,3-dioxo-2,3-dihydro-1H-indene-2-carboxylate (3):

[0189] Ethyl acetoacetate (42 mL, 0.31 mol) and Ac2O (48.5 mL, 0.52 mol) were added at ambient temperature to a suspension of anhydrous 2 (50 g, 0.26 mol) in dry DCM (260 mL). Et3N (108 mL, 0.78 mol) was added dropwise to this suspension over a period of 30 minutes at room temperature and filled. The mixture was stirred at the same temperature for more than 15 minutes, after which the DCM was evaporated. Subsequently, the obtained crude material was dissolved in 2 liters of water and cooled to 0°C. The mixture was secured with an overhead stirrer, and 300 mL of 2 N HCl was added while maintaining a temperature below 0°C under vigorous stirring conditions. The mixture was stirred at 0°C for more than 15 minutes, then filtered through a Buckner funnel and washed with ice water (500 mL). After that, the product was obtained by air drying it for 3 days.

[0190] 4-nitro-1H-indene-1,3(2H)-dion (4):

[0191] Ethyl 4-nitro-1,3-dioxo-2,3-dihydro-1H-indene-2-carboxylate 3 (272.5 g, 1.04 mol) was obtained in 1 liter of MeCN:water (20:1, 1.0 M). This suspension was slowly filled with TFA (60 mL, 1.14 mol) at room temperature and heated to 50°C. After 4 hours, the reaction mixture was concentrated over a rotary evaporator until approximately 100 mL of solvent remained. Subsequently, the precipitated solid was filtered through a Buckner funnel and washed with (1:1) CHCl3:hexane. This yielded a solid product, and the filtrate was concentrated again to obtain more product in a second crop.

[0192] Reaction Equation 2:

[0193]

[0194] 2,2-dihydroxy-4-nitro-1H-indene-1,3(2H)-dione (5):

[0195] 4-nitro-1H-indene-1,3(2H)-dione (4) (250 g, 1.31 mol) was obtained from 1,4-dioxane (2 liters) and AcOH (200 ml). SeO2 (291 g, 2.62 mol) was added to this at room temperature and maintained under reflux at 110°C for the next 4 hours. This was stirred at room temperature for the next 12 hours. Afterward, it was packed with 500 g to 600 g of CELITE. It was stirred well and filtered across a CELITE pad. The residue was washed with ethyl acetate (300 to 500 mL). The obtained filtrate was concentrated to obtain a crude mass, which was then used as is in the next step.

[0196] 4b,9b-dihydroxy-7-isopropyl-4-nitro-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one (7):

[0197] 2,2-dihydroxy-4-nitro-1H-indene-1,3(2H)-dione 5 (crude material, 1.31 mol) was obtained in glacial AcOH (2 liters), packed with 3-isopropylphenol 6 (196 g, 1.44 mol), and maintained under reflux for the next 10 hours. Afterward, the mixture was fully concentrated and purified by silica gel column chromatography (30% EA in hexane) to obtain a pure product.

[0198] 9b-chloro-4b-hydroxy-7-isopropyl-4-nitro-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one (8):

[0199] 4b,9b-dihydroxy-7-isopropyl-4-nitro-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one 7 (50 g, 0.147 mol) was obtained in DCM (500 mL), and this suspension was charged with oxalyl chloride (1.2 eq) in a single lot. Then, it was slowly charged with DMF (50 mL). After that, the reaction mixture was left to be stirred at room temperature for the next 6 hours. It was quenched with water (500 mL) and the layers were separated. The aqueous layer was extracted with DCM (300 mL × 2). The combined organic layer was washed with water (300 mL) and brine (300 mL). This was dried and concentrated over sodium sulfate to obtain the crude material, and then purified over a short silica pad (30% ethyl acetate in hexane) to obtain the pure product. mp: 1 H-NMR (300 MHz, CDCl3): δ 1.18 (dd, J = 3.6 Hz, J = 6.9 Hz, 6H), 2.84 (sept, J = 6.9 Hz, 1H), 6.34 (s, 1H), 6.70 (s, 1H), 6.94 (dd, J = 1.0 Hz, J = 7.8 Hz, 1H), 7.45 (d, J = 7.8 Hz, 1H), 7.81-7.83 (m, 1H), 8.21 (m, 1H), 8.52 (m, 1H).

[0200] 9b-amino-4b-hydroxy-7-isopropyl-4-nitro-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one (9):

[0201] 9b-chloro-4b-hydroxy-7-isopropyl-4-nitro-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one 8 (36.0 g, 0.1 mol) was obtained in THF (350 mL) and cooled to -40°C. A 2.0 M solution of NH3 in IPA (100 mL, 0.20 mol) was added to this using a dropping funnel, and the temperature was maintained below -20°C. The reaction mixture was monitored at -20°C for 1 hour, then heated to room temperature. It was stirred at room temperature until the reaction was complete, and then fully concentrated. The crude material was obtained in ethyl acetate (500 mL) and washed with water (200 mL × 2) and brine (100 mL). This was dried over anhydrous Na2SO4 and concentrated to obtain crude material, which was then purified over a short silica pad to obtain a pure product. 1 H-NMR (300 MHz, CDCl3) δ 1.18 (d, J = 6.9 Hz, 6H), 2.84 (sept, J = 6.9 Hz, 1H), 3.46 (s, 1H), 6.25 (s, 1H), 6.74 (s, 2H), 6.90 (dd, J = 1.2 Hz, J = 7.8 Hz, 1H), 7.55 (d, J = 7.8 Hz, 1H), 7.77 (t, J = 8.1 Hz, 1H), 8.22 (dd, J = 1.2 Hz, J = 8.4 Hz, 1H), 8.52 (dd, J = 1.2 Hz, J = 8.1 Hz, 1H).

[0202] N-(4b-hydroxy-7-isopropyl-4-nitro-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (10):

[0203] 9b-amino-7-cyclopropyl-4b-hydroxy-4-nitro-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one 9 (950 mg, 2.80 mmol) was obtained in AcOH (10 mL, 0.1 M), to which acetic anhydride (0.263 mL, 2.8 mmol) was added at ambient temperature. The mixture was heated at 80°C for 30 minutes. The reaction mixture was concentrated and obtained in EA (100 mL). It was washed with water (30 mL) and brine (30 mL). It was dried and concentrated in anhydrous Na2SO4. The obtained crude material was purified by silica gel column chromatography (30-40% EA in hexane) to obtain a pure product.

[0204] N-(1-amino-4b-hydroxy-7-isopropyl-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (11):

[0205] N-(4b-hydroxy-7-isopropyl-4-nitro-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide 10 (500 mg, 1.3 mmol) was obtained in EtOH:water (10:1, 15 mL, 0.1 M), and Fe powder (0.219 mg, 3.92 mmol) was added to it. This was charged with a catalytic amount of concentrated HCl (3 drops) and refluxed at 90°C for the next 3 hours. The reaction mixture was filtered through CELITE under high temperature conditions, and the residue was washed with EA. It was concentrated and obtained in EA (250 mL). It was washed with water (100 mL) and brine (100 mL). It was dried over anhydrous Na2SO4 and concentrated. The obtained crude material was purified through silica gel column chromatography (1:1 = EA : hexane) to obtain a pure product.

[0206] N-((4bR,9bR)-1-amino-4b-hydroxy-7-isopropyl-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (12) and N-((4bS,9bS)-1-amino-4b-hydroxy-7-isopropyl-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (13):

[0207] N-(1-amino-4b-hydroxy-7-isopropyl-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (560 mg) was purified by chiral chromatography using an AD column (SFC=100 ml / min, CO2 / EtOH=70 / 30, 236 bar) to obtain 243 mg of N-((4bR,9bR)-1-amino-4b-hydroxy-7-isopropyl-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide 12 as (peak 2, tR 4.33 min); 1H NMR (500 MHz, methanol-d4) δ 7.41-7.50 (m, 1H), 7.32-7.40 (m, 1H), 6.94-7.03 (m, 1H), 6.79-6.91 (m, 1H), 6.58-6.74 (m, 2H), 2.77-2.94 (m, 1H), 1.96-2.05 (m, 3H), 1.12-1.26 (m, 6H) and 246 mg of N-((4bS,9bS)-1-amino-4b-hydroxy-7-isopropyl-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide 13 (peak 1. Obtained as tR 2.30 min); 1H NMR (400 MHz, methanol-d4) δ: 7.39-7.46 (m, 1H), 7.35 (br d, J=7.8 Hz, 1H), 6.97 (br d, J=7.3 Hz, 1H), 6.84 (br d, J=7.6 Hz, 1H), 6.61-6.69 (m, 2H), 2.82 (dt, J=13.6, 6.8 Hz, 1H), 1.98 (s, 3H), 1.17 (dd, J=6.9, 1.6 Hz, 6H).

[0208] Examples 3 to 5: N-(1-amino-4b-hydroxy-10-oxo-7-(trifluoromethyl)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (19); N-((4bR,9bR)-1-amino-4b-hydroxy-10-oxo-7-(trifluoromethyl)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (20) and N-((4bS,9bS)-1-amino-4b-hydroxy-10-oxo-7-(trifluoromethyl)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (21)

[0209]

[0210] N-(1-amino-4b-hydroxy-10-oxo-7-(trifluoromethyl)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (19) (500 mg) was purified by chiral chromatography using an AD column (SFC=100 ml / min, CO2 / IPA=80 / 20, 226 bar) to obtain N-((4bR,9bR)-1-amino-4b-hydroxy-10-oxo-7-(trifluoromethyl)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (20) as (peak 2, tR 4.50 min); 1H NMR (500 MHz, methanol-d4) δ 7.58-7.70 (m, 1H), 7.42-7.53 (m, 1H), 7.26 (br d, J=7.80 Hz, 1H), 6.97-7.11 (m, 2H), 6.67-6.83 (m, 1H), 2.02 (s, 3H) and N-((4bS,9bS)-1-amino-4b-hydroxy-10-oxo-7-(trifluoromethyl)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (21) was obtained as (peak 1, tR 2.49 min); 1H NMR (500 ㎒, methanol-d4) δ: -1.13 (br d, J=7.6 ㎐, 1H), -1.29 (br t, J=7.6 ㎐, 1H), -1.50 (br d, J=7.3 ㎐, 1H), -1.79--1.69 (m, 2H), -2.09--1.95 (m, 1H), -6.75 (s, 3H).

[0211] Reaction Equation 3:

[0212]

[0213] 4b,9b-dihydroxy-4-nitro-7-(trifluoromethyl)-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one (15):

[0214] SeO2 (48.8 g, 0.44 mol) was added to a solution of 4-nitro-1H-indene-1,3(2H)-dione 4 (42.1 g, 0.22 mol) in 1,4-dioxane:AcOH (10:1, 330 mL, 0.6 M). The resulting solution was refluxed at 130°C for 3 hours. Afterward, it was cooled and filtered through CELITE using EA (approx. 200 mL). The filtrate was fully concentrated, and crude 2,2-dihydroxy-4-nitro-1H-indene-1,3(2H)-dione 5 was obtained in MeSO3H (350 mL, 0.6 M). To this, 3-(trifluoromethyl)phenol 14 (29 mL, 0.24 mol) was added dropwise, and the mixture was stirred at room temperature (30°C) for the next 24 hours. Afterward, the reaction material was quenched in ice water (1500 mL) and the solid was filtered. The residue was dissolved in EA (500 mL) and washed with water (200 mL) and brine (200 mL). This was dried over anhydrous Na2SO4 and concentrated into a crude material. The crude material was purified by silica gel column chromatography (10-40% EA in hexane with 5-10% DCM) to obtain a pure product.

[0215] 9b-chloro-4b-hydroxy-4-nitro-7-(trifluoromethyl)-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one (16):

[0216] 4b,9b-dihydroxy-4-nitro-7-(trifluoromethyl)-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one 15 (25 g, 68 mmol) was obtained in DCM (270 mL, 0.25 M) and charged with oxalyl chloride (7.1 mL, 82 mmol) at room temperature. DMF (26 mL, 340 mmol) was slowly added to this mixture and stirred at ambient temperature (20°C). Afterward, the reaction mixture was stirred at room temperature (20°C) for the next 6 hours. The reaction was recharged with oxalyl chloride (1.8 mL, 0.3 eq) and stirred for the next 12 hours. The reaction mixture was diluted with water (approx. 300 mL). The aqueous layer was extracted with DCM (approx. 300 mL x 2). The combined organic layer was washed with water (approx. 300 ml) and brine (approx. 300 ml). It was dried over anhydrous Na2SO4 and concentrated to obtain a crude product. The crude material was purified by silica gel column chromatography (10-25% EA in hexane) to obtain a pure product.

[0217] 9b-amino-4b-hydroxy-4-nitro-7-(trifluoromethyl)-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one (17):

[0218] 9b-chloro-4b-hydroxy-4-nitro-7-(trifluoromethyl)-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one 16 (20.5 g, 53 mmol) was prepared in THF (350 mL, 0.15 M) and cooled to -40°C. To this, 2.0 M NH3 in IPA (65 mL, 0.13 mol) was added dropwise over a period of 10 minutes. Subsequently, the reaction mixture was left to be stirred at -40°C for the next 3 hours. Afterward, it was diluted with EA (about 200 to 300 mL) and washed with 10% brine (about 200 mL × 2). The organic layer was dried over anhydrous Na2SO4 and concentrated to obtain the crude product.

[0219] N-(4b-hydroxy-4-nitro-10-oxo-7-(trifluoromethyl)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (18):

[0220] 9b-amino-4b-hydroxy-4-nitro-7-(trifluoromethyl)-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one 17 (21 g, 50 mmol) was obtained in ice AcOH (250 mL, 0.2 M) and immediately charged with Ac2O (9.5 mL, 0.1 mol). Subsequently, the reaction mixture was heated at 80°C for the next 60 minutes. The reaction mixture was concentrated to obtain the crude material. The crude material was immediately purified by silica gel column chromatography (20-40% EA in Hx with 10% DCM as a co-solvent) to obtain the pure product.

[0221] N-(1-amino-4b-hydroxy-10-oxo-7-(trifluoromethyl)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (19):

[0222] N-(4b-hydroxy-4-nitro-10-oxo-7-(trifluoromethyl)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide 18 (9.0 g, 22 mmol) was prepared in EtOH:water (10:1, 110 mL, 0.2 M), charged with Fe powder (3.7 g, 66 mmol), and then charged with HCl (0.5 mL). This was refluxed at 90°C for the next 3 hours. The reaction mixture was filtered through CELITE under heated conditions using high-temperature EA (about 50 to 100 mL). The filtrate was concentrated and obtained in EA (about 600 to 800 mL) and washed with water (400 mL). The aqueous layer was extracted with EA (about 200 mL × 2). The combined organic layer was washed with water (approx. 300 mL) and brine (approx. 200 mL). It was dried and concentrated in anhydrous Na2SO4 (approx. 100 to 150 mL). Afterward, the precipitated solid was thoroughly sonicated and filtered to obtain a pure product. The filtrate was concentrated to obtain a crude material. The crude material was purified by silica gel column chromatography (20-50% EA in Hx with DCM as an additive) to obtain an additional amount of pure product. 1H-NMR (300 ㎒, CD3OD) δ 2.0 (s, 3H), 6.74 (s, 1H), 7.00-7.02 (m, 2H), 7.24 (d, J = 7.8 ㎐, 1H), 7.43-7.48 (m, 1H), 7.61 (d, J = 7.8 ㎐, 1H). LCMS: 378.6 [M + H] + .

[0223]

[0224] N-((4bR,9bR)-1-amino-4b-hydroxy-10-oxo-7-(trifluoromethyl)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (20)

[0225] N-(1-amino-4b-hydroxy-10-oxo-7-(trifluoromethyl)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (19) (500 mg) was purified by chiral chromatography using an AD column (SFC=100 ml / min, CO2 / IPA=80 / 20, 226 bar) to obtain 202 mg of N-((4bR,9bR)-1-amino-4b-hydroxy-10-oxo-7-(trifluoromethyl)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (20) as (peak 2, tR 4.50 min); 1H NMR (500 MHz, methanol-d4) δ 7.58-7.70 (m, 1H), 7.42-7.53 (m, 1H), 7.26 (br d, J=7.80 Hz, 1H), 6.97-7.11 (m, 2H), 6.67-6.83 (m, 1H), 2.02 (s, 3H) and 205 mg of N-((4bS,9bS)-1-amino-4b-hydroxy-10-oxo-7-(trifluoromethyl)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (21) was obtained as (peak 1, tR 2.49 min); 1H NMR (500 ㎒, methanol-d4) δ: -1.13 (br d, J=7.6 ㎐, 1H), -1.29 (br t, J=7.6 ㎐, 1H), -1.50 (br d, J=7.3 ㎐, 1H), -1.79--1.69 (m, 2H), -2.09--1.95 (m, 1H), -6.75 (s, 3H).

[0226] Example 6: N-((4bR,9bR)-1-amino-7-bromo-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (31)

[0227]

[0228] Reaction Equation 4:

[0229]

[0230] 7-bromo-4b,9b-dihydroxy-4-nitro-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one (23):

[0231] 4-nitro-1H-indene-1,3(2H)-dione 4 (10.0 g, 52.3 mmol) was obtained in AcOH:dioxane (1:10, 105 mL, 0.5 M). This was charged with SeO2 (12.77 g, 115.1 mmol) and refluxed at 105-110°C for 5 hours. Afterward, the reaction mixture was filtered through CELITE under high temperature conditions, and the volatile products were concentrated to obtain crude 2,2-dihydroxy-4-nitro-1H-indene-1,3(2H)-dione 5. Subsequently, this crude product was obtained in ice AcOH (210 mL, 0.25 mmol), charged with 3-bromophenol 22 (9.96 g, 57.5 mmol), and maintained under reflux for the next 12 hours. The reaction material was concentrated and obtained in EA (500 to 600 mL). This was filtered through CELITE, and the residue was washed with EA. The filtrate was washed with water (200 mL × 2) and brine (100 mL). This was dried and concentrated in anhydrous Na2SO4 to obtain a crude product. The crude material was purified twice by silica gel column chromatography (35-40% EA in hexane) to obtain a pure product.

[0232] 7-bromo-9b-chloro-4b-hydroxy-4-nitro-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one (24):

[0233] 7-bromo-4b,9b-dihydroxy-4-nitro-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one 23 (39.5 g, 0.105 mol) was obtained in DCM (520 mL, 0.2 M) and charged with oxalyl chloride (11 mL, 0.13 mmol) at room temperature. DMF (40 mL, 0.53 mol) was slowly added to this (0.05 mL / min over 30 minutes, then 0.1 mL / min over 30 minutes, then the remainder), and stirred at ambient temperature (30°C). Afterward, the reaction mixture was stirred at rt (20°C) for the next 12 hours. The reaction mixture was diluted with water (approx. 300 mL). The aqueous layer was extracted with DCM (approx. 500 mL × 2). The combined organic layer was washed with water (approx. 300 ml) and brine (approx. 300 ml). It was dried in anhydrous Na2SO4 and concentrated to obtain a crude product. The crude material was purified by silica gel column chromatography (10-30% EA in hexane) to obtain a pure product.

[0234] 9b-amino-7-bromo-4b-hydroxy-4-nitro-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one (25):

[0235] 9b-chloro-4b-hydroxy-4-nitro-8-(trifluoromethyl)-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one 24 (21.2 g, 53.4 mmol) was obtained in THF (530 mL, 0.1 M) and cooled to -40°C. This was charged at the same temperature into 2.0 M NH3 in IPA (54 mL, 0.11 mmol) and stirred for the next 3 hours. The reaction mixture was diluted with water (approx. 150 mL) and brine (150 mL). The aqueous layer was extracted with EA (approx. 300 mL × 2). The combined organic layer was washed with brine (approx. 100 mL). This was dried over anhydrous Na2SO4 and concentrated to obtain the crude product. The crude material was purified by silica gel column chromatography (20-30% EA in hexane with 20% DCM as a co-solvent) to obtain a pure product.

[0236] Tert-butyl (7-bromo-4b-hydroxy-4-nitro-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)carbamate (26):

[0237] Boc anhydride (8.74 g, 40 mmol) and molecular I2 (0.69 g, 2.67 mmol) were added to a solution of a racemic mixture of 9b-amino-7-bromo-4b-hydroxy-4-nitro-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one 25 (10.1 g, 31 mmol) in THF (5.0 mL, 5.0 M), and stirred at rt (30°C) for the following 72 hours. The reaction mixture was concentrated and purified. The crude material was purified by silica gel column chromatography (10%-30% EA in hexane with 5-10% DCM) to obtain a pure product.

[0238] Tert-butyl (1-amino-7-bromo-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)carbamate (27):

[0239] A mixture of racemic tert-butyl (7-bromo-4b-hydroxy-4-nitro-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)carbamate 26 (10.3 g, 21.5 mmol) was obtained in EtOH:water (10:1, 110.0 mL, 0.20 M), charged with Fe powder (3.57 g, 63.9 mmol), and then charged with concentrated HCl (0.8 mL, cat.). This was refluxed at 90°C for the next 3 hours. The reaction mixture was filtered through CELITE under heated conditions using high-temperature EA (50 to 100 mL). The filtrate was concentrated and obtained in EA (about 1000 to 1200 mL), and washed with water (about 300 to 500 mL) and brine (about 300 mL). This was dried over anhydrous Na2SO4 and concentrated to obtain a crude material. The crude material was purified by silica gel column chromatography (10-30% EA in hx) to obtain a pure product.

[0240] Tert-butyl ((4bR,9bR)-1-amino-7-bromo-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)carbamate (28) and tert-butyl ((4bR,9bR)-1-amino-7-bromo-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)carbamate (29):

[0241] Tert-butyl (1-amino-7-bromo-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)carbamate (27) was purified by chiral chromatography using (AD column, HPLC=20 ml / min, heptane / EtOH=70 / 30, 724 psi) to obtain tert-butyl ((4bR,9bR)-1-amino-7-bromo-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)carbamate (28) as (peak 2, tR 15.59 min); 1H NMR (500 MHz, methanol-d4) δ: 7.48 (br t, J=7.7 Hz, 1H), 7.37 (br s, 1H), 7.11 (br s, 1H), 7.02 (br d, J=7.1 Hz, 1H), 6.95 (s, 1H), 6.72 (br s, 1H), 1.42 (br s, 5H), 1.13 (br s, 4H) LCMS: 447.2 / 449.2 [M + H] + And tert-butyl ((4bS,9bS)-1-amino-7-bromo-4b-hydroxy-10-oxo-9b,10-dihydro-4bH-indeno[1,2-b]benzofuran-9b-yl)carbamate (29) was obtained (peak 1, tR 8.97 min); 1H NMR (500 MHz, methanol-d4) δ: 7.48 (br t, J=7.6 Hz, 1H), 7.37 (br s, 1H), 7.11 (br s, 1H), 7.02 (br d, J=6.9 Hz, 1H), 6.95 (s, 1H), 6.72 (br s, 1H), 1.42 (br s, 5H), 1.13 (br s, 4H) LCMS: 447.2 / 449.2 [M + H] +

[0242] (4bR,9bR)-1,9b-diamino-7-bromo-4b-hydroxy-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one (30):

[0243] Tert-butyl ((4bR,9bR)-1-amino-7-bromo-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)carbamate 28 (112 mg, 0.25 mmol) was obtained from DCM (2.5 mL, 0.1 M) and immediately charged with 4.0 M HCl in dioxane (0.63 mL, 2.50 mmol). Afterward, the reaction mixture was stirred at rt (20°C) for the next 6 hours. The reaction mixture was diluted with EA (50 mL) and vigorously stirred with saturated NaHCO3 (20 mL) for 5-10 minutes. The layers were separated, and the aqueous layer was extracted with EA (30 mL × 2). The combined organic layer was washed with water (20 mL) and brine (20 mL). This was dried and concentrated in anhydrous Na2SO4 to obtain the product. The crude material was used as is in the next step without further purification.

[0244] N-((4bR,9bR)-1-amino-7-bromo-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (31):

[0245] (4bR,9bR)-1,9b-diamino-7-bromo-4b-hydroxy-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one 30 (76 mg, 0.22 mmol) was obtained in ice AcOH (2.2 mL, 0.1 M) and immediately charged with Ac2O (0.03 mL, 1.2 mmol). Subsequently, the reaction mixture was heated at 80°C for the next 30 minutes. Afterward, 10 mL of 2 N HCl (aq.) was added, and the mixture was stirred at 80°C for the next 2 hours. The reaction mixture was concentrated to obtain the crude material. The crude material was immediately purified by silica gel column chromatography (20-50% EA in Hx with 1-2% MeOH as a co-solvent) to obtain the pure product. 1 H-NMR (300 MHz, MeOD) δ 7.50-7.40 (br, 1H), 7.40-7.25 (br, 1H), 7.10 (d, J= 7.4 Hz, 1H), 7.05-6.85 (m, 2H), 6.69 (br, 1H), 1.99 (s, 3H).

[0246]

[0247] Examples 7 to 9: N-(1-amino-7-cyclopropyl-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (40); N-((4bR,9bR)-1-amino-7-cyclopropyl-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (41) and N-((4bS,9bS)-1-amino-7-cyclopropyl-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (42)

[0248]

[0249] Reaction Equation 5:

[0250]

[0251] (3-bromophenoxy)(tert-butyl)dimethylsilane (32):

[0252] 3-Bromophenol 22 (2.08 g, 12 mmol) was obtained in dry DCM (40 mL, 0.3 M). This was charged with TBDMS-Cl (2.0 g, 13 mmol). Subsequently, it was charged with imidazole (1.37 g, 20 mmol) and stirred at room temperature for the next 15 hours. The reaction mixture was immediately filtered, and the residue was washed with DCM. The filtrate was concentrated, and the obtained crude material was purified by silica gel column chromatography (0-5% EA:hexane) to obtain a pure product. 1 H-NMR (300 MHz, CDCl3) δ 0.20 (s, 6H), 0.97 (s, 9H), 6.74-6.78 (m, 1H), 7.00 (s, 1H), 7.07-7.09 (m, 1H).

[0253] Tert-butyl(3-cyclopropylphenoxy)dimethylsilane (34):

[0254] (3-bromophenoxy)(tert-butyl)dimethylsilane 32 (430 mg, 1.5 mmol) was obtained in toluene:water (pre-purged with nitrogen) (7.33 mL, 0.2 M). This was charged with cyclopropaneboronic acid 33 (154 mg, 1.8 mmol). Subsequently, it was charged with PCy3 (42 mg, 0.15 mmol), K3PO4 (1.1 g, 5.24 mmol), and Pd(OAc)2 (17 mg, 0.07 mmol). Afterward, it was refluxed at 110°C for the following 3 hours. The reactants were passed through CELITE and washed with ether. The organic layer was washed with water (30 mL) and brine (30 mL). This was dried and concentrated in anhydrous Na2SO4 to obtain a crude material, and purified by silica gel column chromatography (0-5% EA:hexane) to obtain a pure product. 1 H-NMR (300 MHz, CDCl3) δ 0.19 (s, 6H), 0.63-0.68 (m, 2H), 0.89-1.02 (m, 11H), 1.79-1.88 (m, 1H), 6.52-6.54 (m, 1H), 6.59-6.68 (m, 2H), 7.06-7.11 (m, 1H).

[0255] 3-cyclopropylphenol (35)

[0256] Tert-butyl(3-cyclopropylphenoxy)dimethylsilane 34 (1.74 g, 7.0 mmol) was obtained in THF (23 mL, 0.3 M), and 1.0 M TBAF (9.1 mL, 9.1 mmol) was added to it. The mixture was stirred at room temperature for 75 minutes. The reaction mixture was concentrated and obtained in EA (200 mL). It was washed with saturated NH4Cl (50 mL), water (50 mL), and brine (50 mL). It was dried and concentrated in anhydrous Na2SO4. The obtained crude material was purified by silica gel column chromatography (5% EA in hexane) to obtain a pure product. 1H-NMR (500 MHz, CDCl3) δ 0.69-0.73 (m, 2H), 0.95-0.99 (m, 2H), 1.85-1.90 (m, 1H), 4.71 (br, 1H), 6.56-6.57 (m, 1H), 6.63 (dd, J = 2.5 Hz, J = 8.0 Hz, 1H), 6.70 (d, J = 8.0 Hz, 1H), 7.13-7.16 (m, 1H, ArH).

[0257] 7-cyclopropyl-4b,9b-dihydroxy-4-nitro-4b,9b-dihydro-10H-indeno[1,2-b]benzofuran-10-one (36):

[0258] 2,2-dihydroxy-4-nitro-1H-indene-1,3(2H)-dione 5 (1.80 g, 8.0 mmol) and 3-cyclopropylphenol 35 (1.1 g, 8.0 mmol) were refluxed in ice AcOH (40 mL, 0.2 M) for 2 hours. The reaction mixture was concentrated and dissolved in EA (200 mL). This was washed with water (50 mL) and brine (50 mL). It was dried and concentrated in anhydrous Na2SO4. Subsequently, the obtained crude material was leached in DCM:hexane (approx. 50 mL), and the resulting solid was sonicated. This was filtered to obtain a pure product. The filtrate was concentrated again and purified by silica gel column chromatography (30% EA in hexane) to obtain the remainder product. 1 H-NMR (300 MHz, CD3OD) δ 0.65-0.67 (m, 2H), 0.95-0.99 (m, 2H), 1.81-1.86 (m, 1H), 3.67 (br, 1H), 6.22 (br, 1H), 6.50 (s, 1H), 6.77 (d, J = 8.0 Hz, 1H), 7.43 (d, J = 8.0 Hz, 1H), 7.78-7.82 (m, 1H), 8.18 (d, J = 7.5 Hz, 1H), 8.50 (d, J = 8.0 Hz, 1H).

[0259] 9b-chloro-7-cyclopropyl-4b-hydroxy-4-nitro-4b,9b-dihydro-10H-indeno[1,2-b]benzofuran-10-one (37):

[0260] 7-cyclopropyl-4b,9b-dihydroxy-4-nitro-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one 36 (1.70 g, 5.0 mmol) was obtained from DCM (20 mL, 0.25 M) and filled with oxalyl chloride (0.52 mL, 6.0 mmol). Afterward, it was slowly filled with DMF (2 mL). After 3 hours, additional oxalyl chloride (0.08 mL) was added. Then, the reaction mixture was stirred at room temperature for the next 30 minutes. The reaction mixture was diluted with DCM to 200 mL. It was washed with water (100 mL × 2). Afterward, it was washed with saturated brine (100 mL) and dried over anhydrous Na2SO4. This was concentrated to obtain a crude substance, and purified through silica gel column chromatography (10-15% EA in Hx) to obtain a pure product. 1 H-NMR (300 MHz, CDCl3) δ 0.61-0.68 (m, 2H), 0.93-0.99 (m, 2H), 1.81-1.87 (m, 1H), 6.28 (br, 1H), 6.49 (s, 1H), 6.78 (d, J = 8.1 Hz, 1H, ArH), 7.39 (d, J = 8.1 Hz, 1H), 7.78-7.823 (m, 1H), 8.19 (d, J = 7.5 Hz, 1H), 8.49 (d, J = 8.1 Hz, 1H).

[0261] 9b-amino-7-cyclopropyl-4b-hydroxy-4-nitro-4b,9b-dihydro-10H-indeno[1,2-b]benzofuran-10-one (38):

[0262] 9b-chloro-7-cyclopropyl-4b-hydroxy-4-nitro-4b,9b-dihydro-10H-indeno[1,2-b]benzofuran-10-one 37 (715 mg, 2.0 mmol) was obtained in dry THF (20.0 mL, 0.1 M). This was cooled to -40°C and then charged with 2.0 M NH3 in IPA (2.0 mL, 4.0 mmol). Subsequently, it was stirred at -40°C to -30°C for the next 2 hours. The reaction product was concentrated to half its volume at 25°C and then quenched with water. It was concentrated again to remove all volatiles and obtained in EA (150 mL). It was washed with water (50 mL × 2) and brine (50 mL). It was dried and concentrated over anhydrous Na2SO4. The obtained crude material was purified by passing it through silica gel column chromatography (1:2 = EA : hexane) that had been deactivated with TEA beforehand, to obtain a pure product. 1 H-NMR (300 MHz, CD3OD) δ 0.59-0.64 (m, 2H), 0.85-0.90 (m, 2H), 1.73-1.85 (m, 1H), 6.56 (s, 1H), 6.81 (d, J = 7.8 Hz, 1H), 7.46 (d, J = 7.8 Hz, 1H), 7.77-7.80 (m, 1H), 8.25 (d, J = 7.5 Hz, 1H), 8.58 (d, J = 8.1 Hz, 1H).

[0263] 9b-chloro-7-cyclopropyl-4b-hydroxy-4-nitro-4b,9b-dihydro-10H-indeno[1,2-b]benzofuran-10-one (39):

[0264] 2,4,6-trichlorobenzoic acid (168 mg, 0.75 mmol) was obtained in THF (5 mL, 0.1 M), and NMM (0.083 mL, 0.75 mmol) was added to it at 0°C. This was charged with acetyl chloride (0.054 mL, 0.75 mmol) and stirred for the next 30 minutes at 0°C. Next, this was charged in a single lot with 9b-amino-7-cyclopropyl-4b-hydroxy-4-nitro-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one 38 (170 mg, 0.5 mmol) and stirred for the next 3 hours at 0°C. The reaction product was concentrated and obtained in EA (100 mL). This was washed with water (30 mL) and brine (30 mL). This was dried and concentrated in anhydrous Na2SO4. The obtained crude material was purified by silica gel column chromatography (1:1 = EA : hexane) to obtain a pure product. 1 H-NMR (300 MHz, CDCl3) δ 0.59-0.64 (m, 2H), 0.92-0.98 (m, 2H), 1.77-1.85 (m, 1H), 2.07 (s, 3H), 6.06 (br, 1H), 6.46 (br, 2H), 6.76 (dd, J = 7.8 Hz, J = 1.2 Hz, 1H), 7.38 (d, J = 7.8 Hz, 1H), 7.71-7.76 (m, 1H), 8.19 (d, J = 7.5 Hz, 1H), 8.45 (d, J = 7.8 Hz, 1H).

[0265] N-(1-amino-7-cyclopropyl-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (40):

[0266] N-(7-cyclopropyl-4b-hydroxy-4-nitro-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide 39 (150 mg, 0.39 mmol) was obtained in EtOH : water (10 : 1, 8 mL, 0.05 M), and Fe powder (66 mg, 1.18 mmol) was added to it. This was charged with 2 drops of concentrated HCl, and the reaction was refluxed for the next 2 hours. The reaction mixture was dried over CELITE, and the residue was washed with EA under high temperature conditions. The filtrate was concentrated, and the crude material was obtained in EA (100 mL). This was washed with water (30 mL) and brine (30 mL). This was dried over anhydrous Na2SO4 and concentrated. Afterwards, the obtained crude material was purified through silica gel column chromatography (1:1 = EA : hexane) to obtain a pure product. 1 H-NMR (300 MHz, CD3OD) δ 0.59-0.61 (m, 2H), 0.89-0.92 (m, 2H), 1.79-1.85 (m, 1H), 2.01 (s, 3H), 6.44 (s, 1H), 6.52-6.72 (m, 2H), 6.95-7.02 (m, 1H), 7.29-7.32 (m, 1H), 7.39-7.44 (m, 1H).

[0267]

[0268] N-((4bR,9bR)-1-amino-7-cyclopropyl-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (41) and N-((4bS,9bS)-1-amino-7-cyclopropyl-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (42):

[0269] N-(1-amino-7-cyclopropyl-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (40) (500 mg) was purified by chiral chromatography using (AD column, HPLC=20 ml / min, heptane / EtOH=70 / 30, 722 psi) to obtain N-((4bR,9bR)-1-amino-7-cyclopropyl-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (41) (peak 2, tR 17.95 min); 1H NMR (500 MHz, methanol-d4) δ: 7.43 (br s, 1H), 7.30 (br s, 1H), 6.98 (br s, 1H), 6.60-6.76 (m, 2H), 6.45 (br s, 1H), 1.99 (s, 3H), 1.84 (br s, 1H), 0.91 (br d, J=8.0 Hz, 2H), 0.58-0.66 (m, 2H).

[0270]

[0271] And N-((4bS,9bS)-1-amino-7-cyclopropyl-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (42) was obtained (peak 1, tR 9.16 min); 1H NMR (500 MHz, methanol-d4) δ: 7.44 (br d, J=2.8 Hz, 1H), 7.33 (br d, J=5.7 Hz, 1H), 7.00 (br d, J=1.7 Hz, 1H), 6.73 (br d, J=6.9 Hz, 1H), 6.64-6.71 (m, 1H), 6.47 (br s, 1H), 2.00 (s, 3H), 1.79-1.92 (m, 1H), 0.86-0.99 (m, 2H), 0.57-0.70 (m, 2H).

[0272]

[0273] Examples 10 to 12: N-(1-amino-4b-hydroxy-10-oxo-7-(trifluoromethoxy)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (48); N-((4bR,9bR)-1-amino-4b-hydroxy-10-oxo-7-(trifluoromethoxy)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (49) and N-((4bS,9bS)-1-amino-4b-hydroxy-10-oxo-7-(trifluoromethoxy)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (50)

[0274]

[0275] Reaction Equation 6:

[0276]

[0277] 4b,9b-dihydroxy-4-nitro-7-(trifluoromethoxy)-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one (44):

[0278] 2,2-dihydroxy-4-nitro-1H-indene-1,3(2H)-dione 5 (1.34 g, 6 mmol) was obtained in TFA (24 mL, 0.25 M). 3-(trifluoromethoxy)phenol 43 (1.07 g, 6 mmol) was added to this, and the mixture was stirred at room temperature (30°C) for the next 12 hours. The reaction mixture was concentrated. Afterward, it was obtained in EA (200 mL) and washed with water (100 mL × 2) and brine (100 mL). It was dried over anhydrous Na2SO4 and concentrated to obtain the crude product. Subsequently, the crude product was purified by silica gel column chromatography (1:2 = EA : hexane) to obtain the pure product.

[0279] 9b-chloro-4b-hydroxy-4-nitro-7-(trifluoromethoxy)-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one (45):

[0280] 4b,9b-dihydroxy-4-nitro-7-(trifluoromethoxy)-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one 44 (385 mg, 1.0 mmol) was obtained in DCM (4.0 mL, 0.25 M). Oxalyl chloride (0.103 mL, 1.21 mmol) was added to this, followed by DMF (0.4 mL, 5.0 mmol), and stirred at room temperature (30°C) for 3 hours. The reaction mixture was diluted with DCM (approx. 100 mL), washed with water (50 mL × 2) and brine (50 mL), dried over anhydrous Na2SO4, and concentrated to obtain the crude material. The crude material was purified by silica gel column chromatography (10-15% EA in hexane) to obtain a pure product.

[0281] 9b-amino-4b-hydroxy-4-nitro-7-(trifluoromethoxy)-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one (46):

[0282] 9b-chloro-4b-hydroxy-4-nitro-7-(trifluoromethoxy)-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one 45 (200 mg, 0.50 mmol) was obtained in THF (5.0 mL, 0.1 M). This was cooled to -40°C, charged with 2.0 M NH3 in IPA (0.5 mL, 1.0 mmol), and heated at -10°C for the next 3 hours. The reaction mixture was concentrated, quenched with water (50 mL), and extracted with EA (100 mL). The combined organic layer was washed with water (50 mL) and brine (30 mL), dried over anhydrous Na2SO4, and concentrated to obtain the crude material. The crude material was purified over a short pad (30-40% EA in hexane) of a silica gel column chromatography to obtain a pure product.

[0283] N-(4b-hydroxy-4-nitro-10-oxo-7-(trifluoromethoxy)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (47):

[0284] 2,4,6-trichlorobenzoic acid (89 mg, 0.40 mmol) was obtained from THF (2.0 mL, 0.1 M) and cooled to 0°C. NMM (0.44 mL, 0.40 mmol) was added to this, followed by the addition of AcCl (0.021 mL, 0.30 mmol). The reaction mixture was stirred for 10 minutes, and 9b-amino-4b-hydroxy-4-nitro-7-(trifluoromethoxy)-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one 46 (76 mg, 0.2 mmol) was added. The reaction mixture was stirred at 0°C for the following 1.5 hours. The reaction mixture was concentrated, the residue was quenched with water (50 mL), and extracted with EA (50 mL × 2). The combined organic layer was washed with water (30 mL) and brine (30 mL), dried over anhydrous Na2SO4, and concentrated to obtain a crude material. The crude product was purified by silica gel column chromatography (30-35% EA in hexane) to obtain a pure product.

[0285] N-(1-amino-4b-hydroxy-10-oxo-7-(trifluoromethoxy)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (48):

[0286] N-(4b-hydroxy-4-nitro-10-oxo-7-(trifluoromethoxy)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide 47 (21 mg, 0.05 mmol) was prepared in EtOH:water (10:1, 2.5 mL, 0.02 M) and charged with Fe powder (8.3 mg, 0.15 mmol). This was charged with concentrated HCl (1 drop) and refluxed at 90°C for the following 3 hours. The reaction mixture was filtered through CELITE under high temperature conditions. The residue was washed with EA (approx. 20 mL). It was concentrated and obtained in EA (approx. 50 mL). This was washed with water (approx. 20 mL) and brine (approx. 20 mL). It was dried over anhydrous Na2SO4 and concentrated to obtain the crude material. The crude material was purified directly through reverse-phase HPLC (MeCN:water as eluent) to obtain the crude product. 1 H-NMR (300 MHz, CD3OD) δ 2.01 (s, 3H), 6.71 (s, 1H), 6.77 (d, J = 8.4 Hz, 1H), 6.87 (d, J = 8.4 Hz, 1H), 7.02 (d, J = 7.2 Hz, 1H), 7.45-7.52 (m, 2H).

[0287]

[0288] N-((4bR,9bR)-1-amino-4b-hydroxy-10-oxo-7-(trifluoromethoxy)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (49) and N-((4bS,9bS)-1-amino-4b-hydroxy-10-oxo-7-(trifluoromethoxy)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (50):

[0289] N-(1-amino-4b-hydroxy-10-oxo-7-(trifluoromethoxy)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (48) (200 mg) was purified by chiral chromatography using an AD column (SFC=100 ml / min, CO2 / IPA=85 / 15, 206 bar) to obtain N-((4bR,9bR)-1-amino-4b-hydroxy-10-oxo-7-(trifluoromethoxy)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (49) (peak 2, tR 7.41 min); 1H NMR (500 MHz, methanol-d4) δ: 7.37-7.55 (m, 2H), 7.00 (d, J=7.3 Hz, 1H), 6.85 (br d, J=8.3 Hz, 1H), 6.75 (br d, J=6.9 Hz, 1H), 6.69 (s, 1H), 1.99 (s, 3H).

[0290]

[0291] And N-((4bS,9bS)-1-amino-4b-hydroxy-10-oxo-7-(trifluoromethoxy)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (50) was obtained as (peak 1, tR 4.10 min); 1H NMR (500 MHz, methanol-d4) δ: 7.42-7.52 (m, 2H), 7.00 (d, J=7.3 Hz, 1H), 6.84 (br d, J=8.3 Hz, 1H), 6.75 (br d, J=8.3 Hz, 1H), 6.68 (s, 1H), 1.99 (s, 3H).

[0292]

[0293] Examples 13 to 15: N-(1-amino-7-chloro-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide and N-((4bR,9bR)-1-amino-7-chloro-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide and N-((4bS,9bS)-1-amino-7-chloro-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide

[0294]

[0295] Reaction Equation 7:

[0296]

[0297] 2,2-dihydroxy-4-nitro-1H-indene-1,3(2H)-dione(5):

[0298] Selenium dioxide (2.7 g, 24 mmol) was added to a mixture of 4-nitro-1H-indene-1,3(2H)-dione 4 (2.3 g, 12 mmol) in dioxane: AcoH (20 mL / 2 mL). The resulting reaction mixture was refluxed at 130°C for 3 hours. The reaction mixture was cooled to ambient temperature, diluted with ethyl acetate, passed through a CELITE bed, washed with ethyl acetate, and the solvent evaporated to obtain the crude material. The residue was used as is in the next step without further purification.

[0299] 7-chloro-4b,9b-dihydroxy-4-nitro-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one(52):

[0300] 3-chlorophenol 51 (1.6 g, 12 mmol) was added to a mixture of 2,2-dihydroxy-4-nitro-1H-indene-1,3(2H)-dione 5 (3.7 g, crude material) in acetic acid (20 mL). The resulting reaction mixture was refluxed at 110°C for 12 hours. The reaction mixture was cooled to ambient temperature, diluted with ethyl acetate, passed through a CELITE bed, washed with ethyl acetate, and the solvent evaporated to obtain the crude material. The crude material was purified by silica gel column chromatography (ethyl acetate:hexane) to obtain the product.

[0301] 7,9b-dichloro-4b-hydroxy-4-nitro-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one (53):

[0302] Oxalyl chloride (0.12 mL, 1.44 mmol) was added to a mixture of 52 (400 mg, 1.2 mmol) of 7-chloro-4b,9b-dihydroxy-4-nitro-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one (400 mg, 1.2 mmol) in 6 mL of DCM. DMF (0.4 mL) was added dropwise to the resulting reaction mixture over 1 hour, and the reaction mixture was stirred at ambient temperature for 15 hours. The reaction mixture was diluted with DCM, washed with water (50 mL × 2), the organic layer was washed with a saline solution, dried over Na2SO4, and the solvent was evaporated to obtain the crude material. The crude material was purified by silica gel column chromatography (ethyl acetate:hexane) to obtain the product.

[0303] 9b-amino-7-chloro-4b-hydroxy-4-nitro-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one (54):

[0304] Ammonia in IPA (0.8 mL, 1.6 mmol) was added for 5 minutes to a mixture of 53 (220 mg, 0.63 mmol) of 7,9b-dichloro-4b-hydroxy-4-nitro-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one in THF (3 mL) at -40°C, and the reaction mixture was stirred at -40°C for 2 hours. The reaction mixture was diluted with ethyl acetate and washed with a saline solution (50 mL × 2). Afterward, the organic layer was dried over Na2SO4, and the solvent was evaporated to obtain the (crude) product. The crude material was used as is without purification in the next step.

[0305] N-(7-chloro-4b-hydroxy-4-nitro-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide(55):

[0306] Acetic anhydride (0.07 mL, 0.76 mmol) was added to a solution of 54 (210 mg, 0.63 mmol) of 9b-amino-7-chloro-4b-hydroxy-4-nitro-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one in AcoH (6 mL). The resulting reaction mixture was stirred at 80°C for 1 hour. The reaction mixture was evaporated dry, the residue was dissolved in ethyl acetate (50 mL), the organic layer was washed with water (25 mL x 2), the organic layer was dried over Na2SO4, and the solvent was evaporated to obtain the crude material. The crude material was purified by silica gel column chromatography (ethyl acetate:hexane) to obtain the product.

[0307] N-(1-amino-7-chloro-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (56):

[0308] Fe powder (45 mg 0.8 mmol) and concentrated HCl in a solution of N-(7-chloro-4b-hydroxy-4-nitro-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (100 mg, 0.26 mmol) in EtOH:H2O (9 mL). (1 drop) was added. The resulting reaction mixture was stirred at 90°C for 3 hours. The reaction mixture was passed through a CELITE bed and washed with ethyl acetate. The solvent was evaporated to obtain a residue, and the residue was dissolved in ethyl acetate (100 mL). The organic layer was washed with water (50 mL × 2), dried over Na2SO4, and the solvent was evaporated to obtain a crude material. The crude material was purified by silica gel column chromatography using (ethyl acetate:hexane) to obtain a product. 1 ¹H NMR (300 MHz, methanol- d 4) δ 7.62 - 7.24 (m, 2H), 6.99 (t, J = 8.8 Hz, 2H), 6.82 (d, J = 1.9 Hz, 1H), 6.72 (s, 1H), 2.01 (s, 3H).

[0309]

[0310] N-((4bR,9bR)-1-amino-7-chloro-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (57) and N-((4bS,9bS)-1-amino-7-chloro-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (58):

[0311] N-(1-amino-7-chloro-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (56) (200 mg) was purified by chiral chromatography using an IC column (SFC=100 ml / min, CO2 / MeOH=85 / 15, 206 bar) to obtain N-((4bR,9bR)-1-amino-7-chloro-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (57) (peak 2, tR 7.20 min); 1H NMR (500 MHz, methanol-d4) δ: 7.42-7.49 (m, 1H), 7.39 (br d, J=3.8 Hz, 1H), 6.99 (br d, J=4.5 Hz, 1H), 6.90-6.96 (m, 1H), 6.80 (br s, 1H), 6.66-6.77 (m, 1H), 1.99 (br s, 3H).

[0312]

[0313] And N-((4bS,9bS)-1-amino-7-chloro-4b-hydroxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (58) was obtained (peak 1, tR 4.92 min); 1 H NMR (500 MHz, methanol-d4) δ: 7.47 (t, J=7.8 Hz, 1H), 7.41 (br d, J=8.3 Hz, 1H), 7.01 (d, J=7.3 Hz, 1H), 6.97 (br d, J=8.3 Hz, 1H), 6.82 (s, 1H), 6.76 (br s, 1H), 2.01 (s, 3H).

[0314]

[0315] Examples 16 to 18: N-(1-amino-4b-hydroxy-7-methyl-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide and N-((4bR,9bR)-1-amino-4b-hydroxy-7-methyl-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide and N-((4bS,9bS)-1-amino-4b-hydroxy-7-methyl-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide

[0316]

[0317] Reaction Equation 8:

[0318]

[0319] 4b,9b-dihydroxy-7-methyl-4-nitro-4b,9b-dihydro-10H-indeno[1,2-b]benzofuran-10-one (60)

[0320] 2,2-dihydroxy-4-nitro-1H-indene-1,3(2H)-dione 5 (6.3 g, crude material, 26.15 mmol) was suspended in ice AcOH (44 mL). m-cresol 59 (3.0 mL, 28.77 mmol) was added. The resulting solution was refluxed at 120°C for the next 6 hours and then concentrated. The residue was purified by column chromatography (50% EA in hexane with 50% dichloromethane) and precipitated to obtain a pure product. 1 H NMR (300 MHz, CDCl3) δ 8.51 (dd, J = 8.0, 0.8 Hz, 1H), 8.19 (dd, J = 7.7, 0.9 Hz, 1H), 7.80 (t, J = 7.8 Hz, 1H), 7.45 (d, J = 7.8 Hz, 1H), 6.85 (d, J = 7.8 Hz, 48H), 6.67 (s, 1H), 2.31 (s, 3H).

[0321] 9b-chloro-4b-hydroxy-7-methyl-4-nitro-4b,9b-dihydro-10H-indeno[1,2-b]benzofuran-10-one (61)

[0322] 4b,9b-dihydroxy-7-methyl-4-nitro-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one 60 (5.3 g, 16.91 mmol) was suspended in DCM (67 mL). Oxalyl chloride (1.7 mL, 20.3 mmol) was slowly added (5 min) at ambient temperature, then dried, and DMF (5 mL) was slowly added at ambient temperature. The reaction mixture was stirred overnight at ambient temperature, diluted with DCM, and washed with water. The organic layer was dried over anhydrous Na2SO4, concentrated, purified by column chromatography (25% EA in Hex with 25% DCM), and reprecipitated (DCM / Hex = 1 / 2) to obtain the product. 1 H NMR (300 MHz, CDCl3) δ 8.53 (dd, J = 8.0, 0.9 Hz, 1H), 8.23 ​​(dd, J = 7.7, 1.0 Hz, 1H), 7.83 (t, J = 7.9 Hz, 1H), 7.44 (d, J = 7.9 Hz, 1H), 6.89 (d, J = 7.9 Hz, 1H), 6.68 (d, J = 7.2 Hz, 1H), 6.34 (s, 1H), 2.32 (d, J = 6.1 Hz, 3H).

[0323] 9b-amino-4b-hydroxy-7-methyl-4-nitro-4b,9b-dihydro-10H-indeno[1,2-b]benzofuran-10-one (62)

[0324] 9b-chloro-4b-hydroxy-7-methyl-4-nitro-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one 61 (2.29 g, 6.9 mmol.) was dissolved in dried THF (69 mL), and then a 2.0 M solution of NH3 in IPA (6.9 mL) was added at -40°C. The reaction mixture was heated to -10°C, stirred for 3 hours, diluted with EA, and washed with water. The organic layer was dried over anhydrous Na2SO4, concentrated, purified by column chromatography (33% EA in Hex with 3.3% DCM), reprecipitated (DCM / Hex = 1 / 2), and the product was obtained. 1 H NMR (300 MHz, CDCl3) δ 8.52 (d, J = 8.0 Hz, 1H), 8.14 (d, J = 7.6 Hz, 1H), 7.75 (t, J = 7.8 Hz, 1H), 7.31 (m, 1H), 6.83 (t, J = 8.6 Hz, 1H), 6.67 (s, 1H), 2.30 (d, J = 6.0 Hz, 3H).

[0325] N-(4b-hydroxy-7-methyl-4-nitro-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (63)

[0326] 9b-amino-4b-hydroxy-7-methyl-4-nitro-4b,9b-dihydro-10H-deno[1,2-b]benzofuran-10-one (310 mg, 1.0 mmol.) and acetic anhydride (0.113 mL, 1.2 mmol.) were dissolved in acetic acid (10 mL). The resulting solution was stirred at 80°C for 2 hours. The reaction mixture was cooled to room temperature, then diluted with EA and washed with water. The organic layer was dried over anhydrous MgSO4, concentrated, purified by column chromatography (50% EA in hexane with 5% DCM), and reprecipitated (DCM / Hex = 1 / 2) to obtain the product. 1H NMR (300 MHz, CDCl3) δ 8.47 (dd, J = 8.1, 0.8 Hz, 1H), 8.22 (d, J = 6.9 Hz, 1H), 7.76 (t, J = 7.8 Hz, 1H), 7.42 (d, J = 7.8 Hz, 1H), 6.85 (d, J = 7.9 Hz, 1H), 6.64 (s, 1H), 6.50 (s, 1H), 6.08 (s, 1H), 2.31 (s, 3H), 2.10 (s, 3H).

[0327] N-(1-amino-4b-hydroxy-7-methyl-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (64)

[0328] N-(4b-hydroxy-7-methyl-4-nitro-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide 63 (150 mg, 0.4233 mmol.) was dissolved in ethanol (8 mL). Iron powder (70 mg, 1.27 mmol.), water (0.8 mL), and concentrated HCl (2 drops) were added to this solution. The resulting solution was stirred at 90°C for 2 hours. The reaction mixture was cooled to room temperature and filtered through a CELITE pad. The filtrate was concentrated, then purified by column chromatography (50% to 150% EA in Hex with 5% DCM), reprecipitated with EA, and the product was obtained. 1 H NMR (300 MHz, CDCl3) δ 8.83 (s, 0.5H), 7.52 (m, 1.5H), 7.23 (m, 1.6H), 7.17 (d, J = 7.5 Hz, 0.7H), 6.85 (m, 1H), 6.77 (d, J = 7.8 Hz, 0.7H), 6.66 (m, 1.5H), 6.60 (d, J = 8.1 Hz, 0.7H), 6.54 (brs, 0.3H), 5.76 (d, J= 8.9 Hz, 1H), 5.55 (brs, 1H), 2.29 (s, 2H), 2.26 (s, 1H), 2.06 (s, 3H).

[0329]

[0330] N-((4bR,9bR)-1-amino-4b-hydroxy-7-methyl-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (65) and N-((4bS,9bS)-1-amino-4b-hydroxy-7-methyl-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (66)

[0331] N-(1-amino-4b-hydroxy-7-methyl-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (64) (200 mg) was purified by chiral chromatography using (AD column, HPLC=20 ml / min, heptane / IPA=70 / 30, 759 psi) to obtain N-((4bR,9bR)-1-amino-4b-hydroxy-7-methyl-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (65) (peak 2, tR 12.49 min); 1H NMR (500 mHz, methanol-d4) δ: 7.38-7.47 (m, 1H), 7.28-7.37 (m, 1H), 6.97 (br d, J=6.6 Hz, 1H), 6.79 (br d, J=6.9 Hz, 1H), 6.66 (br d, J=7.3 Hz, 1H), 6.59 (br s, 1H), 2.27 (s, 3H), 1.99 (s, 3H).

[0332]

[0333] And N-((4bS,9bS)-1-amino-4b-hydroxy-7-methyl-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (66) was obtained (peak 1, tR 7.77 min); 1H NMR (500 ㎒, methanol-d4) δ: 7.44 (br s, 1H), 7.34 (br s, 1H), 6.99 (br d, J=5.7 ㎐, 1H), 6.80 (br d, J=6.1 ㎐, 1H), 6.68 (br d, J=4.0 ㎐, 1H), 6.61 (br s, 1H), 2.28 (s, 3H), 2.01 (s, 3H).

[0334]

[0335] Example 19: N-(1-amino-4b-hydroxy-8-methyl-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide.

[0336]

[0337] The procedure for the above compound follows a route similar to that described in Examples 16 to 18, except that p-cresol was used instead of m-cresol to obtain N-(1-amino-4b-hydroxy-8-methyl-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide. 1 H NMR (500 ㎒, methanol-d4) δ: 7.44 (br t, J=7.2 ㎐, 1H), 7.30 (br s, 1H), 7.09 (br d, J=7.8 ㎐, 1H), 6.99 (br d, J=6.9 ㎐, 1H), 6.67 (br t, J=9.2 Hz, 2H), 2.30 (br s, 3H), 2.01 (s, 3H).

[0338]

[0339] Example 20: N-((4bR,9bR)-1-amino-4b-hydroxy-8-methyl-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide.

[0340]

[0341] Racemic mixture N-(1-amino-4b-hydroxy-8-methyl-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (200 mg) was purified by chiral chromatography using an AD column (SFC=100 ml / min, CO2 / EtOH=75 / 25, 226 bar) to obtain the product N-((4bR,9bR)-1-amino-4b-hydroxy-8-methyl-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide as (peak 2, tR 5.43 min); 1H NMR (400 MHz, methanol-d4) δ: 7.42 (br t, J=7.1 Hz, 1H), 7.27 (br s, 1H), 7.06 (br d, J=7.9 Hz, 1H), 6.97 (br d, J=6.8 Hz, 1H), 6.64 (br d, J=8.1 Hz, 2H), 2.28 (s, 3H), 1.98 (s, 3H) and also N-((4bS,9bS)-1-amino-4b-hydroxy-8-methyl-10-oxo-9b,10-dihydro-4bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (peak 1, tR It was obtained as 3.68 minutes.

[0342]

[0343] Example 21: N-(1-amino-4b-hydroxy-10-oxo-7-(trifluoromethyl)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)-N-methylacetamide

[0344]

[0345] The procedure for the above compound followed a route similar to that described in Examples 3 to 5, except that 2.0 M methylamine in THF was used instead of 2.0 M ammonia in IPA to obtain the product N-(1-amino-4b-hydroxy-10-oxo-7-(trifluoromethyl)-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)-N-methylacetamide. 1H NMR (500 MHz, methanol-d4) δ: 7.63 (br d, J=7.6 Hz, 1H), 7.44 (br t, J=7.8 Hz, 1H), 7.31 (br d, J=8.0 Hz, 1H), 7.10 (s, 1H), 6.99 (br d, J=7.3 Hz, 1H), 6.70 (br d, J=7.3 Hz, 1H), 2.88 (s, 3H), 2.19 (s, 3H).

[0346]

[0347] Examples 22 to 24: N-(1-amino-4b-hydroxy-7-methoxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (Example 22), N-((4bR,9bR)-1-amino-4b-hydroxy-7-methoxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (Example 23), and N-((4bS,9bS)-1-amino-4b-hydroxy-7-methoxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (Example 24)

[0348]

[0349] The procedure for the above compound followed a route similar to that described in Examples 16 to 18, except that 3-methoxyphenol was used instead of m-cresol to obtain the racemic mixture N-(1-amino-4b-hydroxy-7-methoxy-10-oxo-9b,10-dihydro-4bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (Example 22) (200 mg), which was purified by chiral chromatography using AD column (HPLC=20 ml / min, heptane / IPA=70 / 30, 723 psi) to obtain the product N-((4bR,9bR)-1-amino-4b-hydroxy-7-methoxy-10-oxo-4b,10-dihydro-9bH-indeno[1,2-b]benzofuran-9b-yl)acetamide As (peak 2, tR 24.20 min); 1H NMR (500 MHz, methanol-d4) δ: 7.39-7.50 (m, 1H), 7.31 (br dd, J=3.1, 2.4 Hz, 1H), 6.92-7.05 (m, 1H), 6.62-6.74 (m, 1H), 6.44-6.57 (m, 1H), 6.34 (s, 1H), 3.72 (s, 3H), 1.99 (s, 3H) (Example 23) and also N-((4bS,9bS)-1-amino-4b-hydroxy-7-methoxy-10-oxo-9b,10-dihydro-4bH-indeno[1,2-b]benzofuran-9b-yl)acetamide (peak 1, tR 9.78 min) (Example 24) was obtained.

[0350]

[0351] The biological activity of the compound of the present invention was measured using the following method.

[0352] Determination of drug efficacy regarding picornavirus cytopathic effect (CPE)

[0353] Inhibition test

[0354] For the assay, HeLa (human cervical cancer cells), MRC-5 (human fetal lung fibroblasts), and RD cells (derived from human rhabdomyosarcoma) were used. For comparison, ribavirin (Riv), pleconaryl (pleco), and BTA-798 (BTA) were used as controls. The reagents were dissolved in 100% dimethyl sulfoxide (DMSO) at concentrations of 10 to 40 mg / ml. The water-soluble reagents were dissolved in PBS(-) solution and stored at -20°C. On the day of the experiment, they were used at 3 to 5 times the concentration in this manner, such that the concentration of dimethyl sulfoxide in each well was 0.5% to 1%.

[0355] Pharmaceutical efficacy was determined using the virus-induced cytopathic effect (CPE) inhibition assay. In this regard, virus-compatible cells were grown in 96-well plates, and then 100 CCID of a viral dilution—either DME supplemented with 2% FBS (DME / 2% FBS) or MEM supplemented with 2% FBS (MEM / 2% FBS)—was introduced into each well of the plate. 50 100 μl of a concentration corresponding to the (50% cell culture infection dose) was inoculated and incubated at 33°C or 37°C for 30 minutes to 1 hour to allow the virus to adsorb onto the cells. After removing the culture medium, aliquots of drug dilutions of various concentrations were added to each well in an amount of 100 μl. Human Rhinovirus (HRV) was grown at 33°C, while other viruses were incubated in a 37°C CO2 incubator for 2 to 3 days. Alternatively, cells were cultured for 2 to 3 days without removing the medium, after which they were added with 50 μl of each drug dilution at twice the concentration, followed by 50 μl of the virus dilution. The viruses were incubated in host HeLa cells in DME / 2% or MEM / 2% FBS at 37°C for 2 to 3 days.

[0356] In the case of HeLa cells, the drug EC 50 It was measured at the (50% maximum effective concentration), which is the drug concentration that induces a response half between the baseline and the maximum using the MTT assay. For RD and MRC-5 cells, CPE was determined using FDA (fluorescein diacetate) or MTT. To determine the effect of drug toxicity on efficacy results, mock infection was also included when inoculated with the virus. Virus-free medium was added to the cell culture, and then subjected to the same treatment as virus-infected cells inoculated with the virus. In other words, after 1 hour of incubation, the medium was removed, and a diluted drug solution was added to the medium one or more times. After incubation for 2 to 3 days, the cells were observed under a microscope, and the drug CC was determined using the MTT assay. 50 The cytotoxicity concentration (50%) was determined, at which 50% of the cells were killed, and in the above assay, the count of surviving cells in the mock-infected wells containing the drug was compared with the count of surviving cells in the control wells not containing the drug. In the FDA hydrolysis assay, FDA was added to each well after removing the medium and incubated for 20 to 30 minutes, and then the fluorescence intensity was measured using a spectrofluorescence meter to determine CPE in the same manner as MTT.

[0357] In other words, the survival rate (survival %) of mock-infected cells for cytotoxicity measurement was calculated using Equation 1 below:

[0358] Cell viability calculated using the formula: Cell Drug = [A (Drug) ― A (Background Solution) / A (Cell Count) ― (Background × 100% Solution)]

[0359] A 100% cell viability indicates no cytotoxicity of the drug, while peak cytotoxicity is reflected by a 0% cell viability. A 50% cytotoxic concentration was defined as the concentration required to reduce the cell count by 50%. These drug concentrations are CC 50 It is represented as follows. A higher value means lower cytotoxicity.

[0360] In addition, the antiviral effect can be calculated using the following mathematical formula 2:

[0361] Antiviral effect = [A (Drug / Virus) ― A (Virus control) / A (Cell control) ― A (Virus control)]

[0362] If the survival rate is 100%, its antiviral effect is 100%, whereas if the survival rate is 0%, it has no antiviral effect. The concentration of a drug that enables a 50% survival rate of virus-infected cells within a well is EC 50 While it can be calculated as such, the lower this value, the better the antiviral effect.

[0363] CC exhibiting cytotoxicity to the compound in some embodiments 50 EC12 showing activity against concentrations and many rhinoviruses belonging to picornaviruses 50 The concentrations are listed in Table 1 below.

[0364] Determination of drug effects against picornavirus using multicycle cytopathic effects (CPE)

[0365] Reduction test

[0366] The drug efficacy against picornavirus was determined using a multicycle CPE reduction assay. The antiviral activity of the compound was initially determined by a CPE reduction assay based on MIS[3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium.

[0367] Specifically, cells grown to confluence in a 96-well plate at 100 50% cell culture infection dose (CCID 50 The cells were infected with the virus. After an adsorption period of 2 hours at 37°C, the virus was removed, and a series of dilutions of the compound were added. The cultures were further incubated at 37°C for 3 days until complete CPE was observed in the infected and uninfected virus control (VC). After removing the medium, 90 μl of cell medium and 10 μl of MTS-phenazine methosulfate (Promega, Leiden, The Netherlands) were added to each well. After an incubation period of 2 hours at 37°C, the optical density (OD) of each well was read at 498 nm using a microplate reader.

[0368] The CPE value % for evaluating antiviral activity was calculated using Equation 3 below:

[0369] % CPE = 100 × [OD (CC) ― OD (Virus + Compound) / OD (CC) ― OD (VC)]

[0370] The CPE value % for measuring the cytotoxicity of the drug was calculated by the following Equation 4:

[0371] % CPE = 100 × [OD (CC) - OD (Virus + Compound) / OD (CC) - OD (Blank)]

[0372] In mathematical formulas 3 and 4 above,

[0373] OD(CC) represents the OD of background cell cultures not induced by viruses or treated by chemicals, and

[0374] OD (VC) represents the OD of a control cell culture induced by a virus but not treated with chemicals, and

[0375] OD(virus + compound) represents the OD of a cell culture infected by a virus that was treated with a concentrated compound, and

[0376] OD(compound) represents the OD of a cell culture treated only with a concentrated compound, and OD(blank) represents the OD of a well to which only the cell culture was added.

[0377] Effective concentration (EC 50 ) represents the drug concentration at which 50% of cells survive due to the CPE of the induced virus, and the cytotoxic concentration (CC 50 ) represents the concentration of the drug at which the compound killed 50% of the cells, and these were calculated by logarithmic interpolation.

[0378] Toxicity concentrations against various viruses for some compounds of the examples (CC 50 ) and effective concentration (EC 50 ) is listed in Table 1 below.

[0379] [Table 1]

[0380]

[0381] As shown in Table 1 above, most compounds according to the present invention have high CC. 50 It is found to have low toxicity as it exhibits a high concentration. In addition, the novel compound according to the present invention has been found to have very high antiviral activity against many rhinoviruses (HRV). In addition, the novel compound according to the present invention has been found to have high antiviral activity against Coxsackievirus B4 (Cox B4) and Poliovirus 1 (PV1).

[0382] Therefore, since the compounds according to the present invention exhibit low cytotoxicity and high antiviral activity against various rhinoviruses, these compounds can be usefully used in pharmacological compositions for preventing or treating diseases caused by picornaviruses to which the virus belongs.

[0383] Accordingly, since the compounds according to the present invention have low cytotoxicity against picornaviruses, including Coxsackievirus, poliovirus, and rhinovirus, and exhibit excellent antiviral activity, they can be effectively used for the prevention or treatment of diseases caused by these viruses, such as polio, acute hemorrhagic conjunctivitis, viral meningitis, hand, foot, and mouth disease, blister disease, hepatitis A, myositis, myocarditis, pancreatitis, diabetes, epidemic myalgia, encephalitis, influenza, herpangina, foot and mouth disease, asthma, chronic obstructive pulmonary disease, pneumonia, sinusitis, or otitis media, as well as respiratory, cardiovascular, and neurological diseases.

[0384] Since compounds represented by the chemical formula according to the present invention, which exist in a mutually equilibrium state, have low cytotoxicity as well as excellent antiviral activity against picornaviruses including Coxsackievirus, Enterovirus, Echovirus, Poliovirus, and Rhinovirus, they can be effectively used as pharmaceutical compositions for the prevention or treatment of viral diseases, such as polio, acute hemorrhagic conjunctivitis, viral meningitis, hand, foot, and mouth disease, blistering disease, hepatitis A, myositis, myocarditis, pancreatitis, diabetes, epidemic myalgia, encephalitis, influenza, herpangina, foot and mouth disease, asthma, chronic obstructive pulmonary disease, pneumonia, sinusitis, or otitis media.

Claims

Claim 1 Human treatment methods.

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