New use

Abstract

The present disclosure provides for the administration of inhibitors of phosphodiesterase 1 (PDE1) for the treatment and prevention of diseases or disorders characterized by inflammation, including methods of treatment and pharmaceutical compositions for use therein.

Description

[Technical Field]

[0001] The relevant technical field relates to the administration of phosphodiesterase 1 (PDE1) inhibitors to promote the resolution of inflammation, for example, through polarization from M1 macrophages to M2 macrophages, as well as to the treatment and prevention of inflammation-related diseases or disorders. [Background technology]

[0002] Eleven families of phosphodiesterases (PDEs) have been identified, but Ca 2+ - Only calmodulin-dependent phosphodiesterase (CaM-PDE) Ca 2+ -These PDEs are activated by calmodulin and have been shown to mediate calcium and cyclic nucleotide (e.g., cAMP and cGMP) signaling pathways. Therefore, these PDEs become activated in a stimulated state when intracellular calcium levels are elevated, leading to increased hydrolysis of cyclic nucleotides. The three known CaM-PDE genes, PDE1A, PDE1B, and PDE1C, are all expressed in central nervous system tissues. In the brain, PDE1A is primarily expressed in the cortex and neostriatum, PDE1B is expressed in the neostriatum, prefrontal cortex, hippocampus, and olfactory tubercle, and PDE1C is more ubiquitous.

[0003] PDE4 is a major cAMP-metabolizing enzyme found in inflammatory and immune cells, making PDE4 inhibitors interesting as anti-inflammatory agents. However, PDE1, which is induced in monocyte-macrophage colony-stimulating factor (GM-CSF), a cytokine, is not thought to play a major role in inflammatory responses. Vinpocetine, a PDE1 inhibitor, has been shown to be anti-inflammatory, but its anti-inflammatory effect is thought to be caused by direct inhibition of the IκB kinase complex (IKK) rather than PDE blockade.

[0004] Macrophages play a central role in maintaining homeostasis and mediating inflammation in the body. Macrophages possess the ability to polarize, expressing different functional programs in response to microenvironmental signals. While there are several activated forms of macrophages, two main groups are designated M1 and M2. M1 macrophages, also called "classically activated macrophages," are activated by LPS and IFN-γ, secreting high levels of IL-12 and low levels of IL-10 to exert pro-inflammatory effects. In contrast, the designation M2, also called "selectively activated macrophages," broadly refers to macrophages that function in constructive processes such as wound healing and tissue repair, and macrophages that block harmful immune system activation by producing anti-inflammatory cytokines such as IL-10. M2 macrophages produce high levels of IL-10 and TGF-β, as well as low levels of IL-12. Long-term presence of M1-type macrophages is detrimental to the body, thus requiring tissue repair and recovery.

[0005] When tissue is exposed to pathogens, circulating inflammatory monocytes are recruited and differentiated into macrophages. Generally, macrophages polarize into the M1 phenotype in the early stages of bacterial infection. When bacteria are recognized by pathogen recognition receptors, macrophages are activated and produce large amounts of pro-inflammatory mediators, including TNFα, IL-1, and nitric oxide (NO), to kill the invading organism and activate adaptive immunity. For example, this mechanism is thought to be involved in the early stages of infections with Salmonella typhi, Salmonella typhimurium, and Listeria monocytogenes, as well as infections with Mycobacterium tuberculosis, Mycobacterium ulcerans, and Mycobacterium avium. If the macrophage-mediated inflammatory response cannot be rapidly controlled, a cytokine storm can form, leading to the pathogenesis of severe sepsis. To counteract the excessive inflammatory response, macrophages either undergo apoptosis or polarize into the M2 phenotype to protect the host from excessive injury and promote wound healing.

[0006] Macrophage polarization is also involved in viral infections, and among them, M2 phenotypic macrophages can suppress inflammation and promote tissue healing. Influenza viruses enhance the phagocytic function of human macrophages, a major characteristic of the M2 phenotype, to eliminate apoptotic cells and promote the resolution of inflammation. In severe acute respiratory syndrome (SARS)-CoV infection, M2 phenotypic macrophages are important for regulating the immune response via STAT-dependent pathways and protecting the host from progression to long-term fibrous lung disease. Furthermore, polynuclear respiratory virus (RSV)-induced bronchiolitis is closely associated with the presence of mixed M1 and M2 macrophages.

[0007] Many viruses adapt to macrophage polarization and induce mechanisms that modulate it. In human monocyte-derived macrophages, HIV-1 infection has been observed to shift cells towards an M1-like state, which correlates with the downregulation of M2 state markers (i.e., CD163, CD206, CCL18, and IL-10) and increased secretion of M1-related chemokines, including CCL3, CCL4, and CCL5.

[0008] Macrophage polarization has also been shown to play an important role in various inflammatory diseases and disorders, particularly non-alcoholic steatohepatitis (NASH), atherosclerosis, metabolic diseases, and systemic lupus erythematosus.

[0009] It had not previously been shown that PDE1 plays a crucial role in mediating the resolution of inflammation or significantly impacts inflammatory diseases. Numerous general inflammatory processes and related diseases and disorders exist, and their mechanisms and actions are still not well understood. Currently, there is a largely unmet need for effective methods to treat inflammation and inflammation-related diseases and disorders, particularly regarding inflammation occurring in the brain. [Brief explanation of the drawing]

[0010] [Figure 1] Figure 1 shows the number of white blood cells detected at the site of inflammation after sterile injury following treatment with compound 1. [Figure 2A] Figure 2A shows the number of macrophages detected at the site of inflammation after sterile injury when treated with compound 1. [Figure 2B] Figure 2B shows the number of macrophages, expressed as a percentage of the total white blood cell count, detected at the site of inflammation after sterile injury following treatment with compound 1. [Figure 3A] Figure 3A shows the number of neutrophils detected at the site of inflammation after sterile injury following treatment with compound 1. [Figure 3B]Figure 3B shows the amount of neutrophils, expressed as a percentage of the total white blood cell count, detected at the site of inflammation after sterile injury following treatment with compound 1. [Figure 4A] Figure 4A shows the amount of M1 macrophages, expressed as a percentage of the total number of macrophages detected at the site of inflammation after sterile injury, when treated with compound 1. [Figure 4B] Figure 4B shows the amount of M2 macrophages, expressed as a percentage of the total number of macrophages detected at the site of inflammation after sterile injury, when treated with compound 1. [Figure 5A] Figure 5A shows the number of M1 macrophages detected at the site of inflammation in the M2-activated state after sterile injury following treatment with compound 1. [Figure 5B] Figure 5B shows the number of M2 macrophages detected at the site of inflammation after sterile injury when treated with compound 1. [Figure 6A] Figure 6A shows the mean fluorescence intensity (MFI) of CD38 expression on macrophage populations detected at inflammatory sites after sterile injury after treatment with compound 1. [Figure 6B] Figure 6B shows the mean fluorescence intensity (MFI) of CD38 expression on macrophage populations detected at inflammatory sites after sterile injury following treatment with compound 1. [Figure 7] Figure 7 shows cytokine production in plasma of test subjects after sterile injury following treatment with compound 1. [Figure 8] Figure 8 shows the number of M1-activated macrophages detected at the site of inflammation after sterile injury following treatment with compound 2. [Figure 9] Figure 9 shows the number of M2-activated macrophages detected at the site of inflammation after sterile injury following treatment with compound 2. [Figure 10] Figure 10 shows the results for compound 1 regarding the motility of BV2 cells in a microglia chemotaxis assay. [Figure 11A] Figure 11A shows the amount of CD80+ macrophages as a percentage of the total number of macrophages detected at the site of inflammation. [Figure 11B] Figure 11B shows the amount of iNOS+ macrophages as a percentage of the total number of macrophages detected at the site of inflammation. [Figure 12A] Figure 12A shows the amount of Arg1+ macrophages as a percentage of the total number of macrophages detected at the site of inflammation. [Figure 12B] Figure 12A shows the amount of CD206+ macrophages as a percentage of the total number of macrophages detected at the site of inflammation. [Overview of the project]

[0011] Surprisingly, the inventors discovered that PDE1 mediates the expression of certain pro-inflammatory cytokines and chemokines, and that PDE1 inhibitors have specific anti-inflammatory effects. In one embodiment, inhibition of PDE1 modulates inflammatory activity in macrophages and reduces the expression of pro-inflammatory genes, thereby providing novel therapeutic methods for various macrophage-mediated disorders and conditions.

[0012] Positive regulation of the inflammatory response in macrophages by elevated intracellular cyclic nucleotide levels offers a promising area for therapeutic intervention. PDE1B is known to be present in monocytes and is involved in macrophage differentiation via growth factor-activated signaling, such as GM-CSF. (Bender AT, et al., Selective up-regulation of PDE1B2 upon monocyte-to-macrophage differentiation, Proc Natl Acad Sci US A. 2005 Jan 11; 102(2): 497-502.) Cyclic guanosine monophosphate (cGMP) has been shown to be an important regulator of the macrophage differentiation pathway. cGMP also plays a role in regulating inflammatory processes such as inducible NO synthase induction and TNFα release. Therefore, significant upregulation of PDE1B may be important in regulating these processes in differentiated macrophages. This suggests that PDE1 inhibitors, such as the PDE1 inhibitors disclosed herein, may prove beneficial in diseases associated with inflammatory disorders related to macrophage activation, for example.

[0013] Accordingly, in one embodiment, the present invention provides the use of various PDE1 inhibitory compounds to treat inflammation and / or inflammation-related diseases or disorders. While not limited to theory, one possible mechanism of this activity is that inhibition of PDE1B may affect macrophage activation in the blood and / or microglia activation in the CNS, such as by reducing M1 activation and the release of pro-inflammatory cytokines, and by promoting the polarization of macrophages to the M2 type through the upregulation of anti-inflammatory cytokines such as IL-10. A discussion of the treatment and prevention of inflammation and / or inflammation-related diseases or disorders such as those related to microglia is found in International Publication WO2018 / 049417 (which, with due attribution, is part of this specification in whole).

[0014] The regulation of M1-to-M2 activation in macrophages is central to inflammatory pathways in many diseases. The role of M1-to-M2 polarization in macrophages is important in many inflammation-related disorders, including: bacterial infections (e.g., infections with Salmonella typhi, Salmonella typhimurium, Listeria monocytogenes, Mycobacterium tuberculosis, Mycobacterium ulcerans, and Mycobacterium avium); viral infections (e.g., African Swine Fever Virus, Classical Swine Fever Virus, Dengue Virus, Foot and Mouth Disease Virus, Human Immunodeficiency Virus). Viruses (HIV) (e.g., HIV1), influenza A virus, porcine circovirus-2, porcine reproductive and respiratory syndrome virus, porcine pseudorabies virus, respiratory syncytial virus, severe acute respiratory syndrome coronavirus, West Nile virus, viral hepatitis (e.g., hepatitis A, hepatitis B, hepatitis C));Parasitic infections (e.g., infections of Taenia crassiceps, Toxoplasma gondii, Leishmania infantum, and Schistosoma mansoni); atopic dermatitis; pneumonia; cardiovascular diseases, e.g., atherosclerosis; obesity and insulin resistance; asthma; pulmonary fibrosis; cardiac obstructive pulmonary disease (COPD); neuropathic pain; stroke; diabetes mellitus; sepsis; non-alcoholic steatohepatitis (NASH); autoimmune hepatitis; systemic lupus erythematosus (SLE); wound healing; pleurisy; peritonitis; and cystic fibrosis.

[0015] Targeted inhibition of PDE1 by the compounds of the present invention is thought to affect macrophage activation and promote the production of anti-inflammatory cytokines and factors involved in the resolution of macrophage-mediated inflammation.

[0016] Accordingly, in one embodiment, the present invention provides a method for promoting the resolution of inflammation for the treatment or prevention of inflammation or inflammation-related diseases, comprising administering a specific inhibitor of phosphodiesterase type I (e.g., a PDE1 inhibitor, e.g., a PDE1B inhibitor) (e.g., a PDE1 inhibitor represented by formulas I, Ia, II, III, IV, V and / or VI as described herein).

[0017] In one embodiment, the present invention provides a method for promoting macrophage activation to an M2 activated state, comprising administering a specific inhibitor of phosphodiesterase type I (e.g., a PDE1 inhibitor, e.g., a PDE1B inhibitor) (e.g., a PDE1 inhibitor represented by formulas I, Ia, II, III, IV, V and / or VI as described herein).

[0018] In one embodiment, the present invention relates to, for example, bacterial infections (e.g., infections caused by Salmonella typhi, Salmonella typhimurium, Listeria monocytogenes, Mycobacterium tuberculosis, Mycobacterium ulcerans, and Mycobacterium avium); viral infections (e.g., African Swine Fever Virus, Classical Swine Fever Virus, Dengue Virus, Foot and Mouth Disease Virus, Human Immunodeficiency Virus (HIV) (e.g., HIV1), Influenza A virus) Viruses, Porcine Circovirus-2, Porcine Reproductive and Respiratory Syndrome Virus, Porcine Pseudorabies Virus, Respiratory Syncytial Virus, Severe Acute Respiratory Syndrome Coronavirus, West Nile Virus, Viral Hepatitis (e.g., Hepatitis A, Hepatitis B, Hepatitis C); Parasitic Infections (e.g., Taenia crassiceps, Toxoplasma gondii, Leishmania infantum, Schistosoma mansoni) Mansoni infections; atopic dermatitis; pneumonia; cardiovascular diseases, e.g., atherosclerosis; obesity and insulin resistance;A method for treating diseases or disorders related to inflammation and / or inflammation and / or microglial function, including asthma; pulmonary fibrosis; cardiac obstructive pulmonary disease (COPD); neuropathic pain; stroke; diabetes mellitus; sepsis; non-alcoholic steatohepatitis (NASH); autoimmune hepatitis; systemic lupus erythematosus (SLE); wound healing; pleurisy; peritonitis; and cystic fibrosis, comprising administering to a patient in need of such treatment an effective amount of the PDE1 inhibitor of the present invention (for example, a PDE1 inhibitor represented by formulas I, Ia, II, III, IV, V and / or VI as described herein), for example, an amount effective in promoting macrophage activation from the M1 activated state to the M2 activated state.

[0019] Further embodiments of the present invention will be shown or become apparent from the following detailed description and the examples herein. [Modes for carrying out the invention]

[0020] Detailed description of the invention Compounds for use in the method of the present invention In one embodiment, the PDE1 inhibitor for use in the therapeutic and prophylactic methods described herein is a substituted 7,8-dihydroimidazo[1,2-a]pyrazolo[4,3-e]pyrimidine-4-one compound and a 7,8,9-trihydro-[1H or 2H]-pyrimido[1,2-a]pyrazolo[4,3-e]pyrimidine-4(5H)-one compound, either in free form or in pharmaceutically acceptable salt form.

[0021] In yet another embodiment, the present invention provides that a PDE1 inhibitor for use in the therapeutic and prophylactic methods described herein is selected from any of the applicant's own publications listed below: U.S. Patent Publication No. 2008-0188492A1, U.S. Patent Publication No. 2010-0173878A1, U.S. Patent Publication No. 2010-0273754A1, U.S. Patent Publication No. 2010-0273753A1, International Publication No. 2010 / 065153, International Publication No. 2 International Publication Nos. 010 / 065151, International Publication Nos. 2010 / 065151, International Publication Nos. 2010 / 065149, International Publication Nos. 2010 / 065147, International Publication Nos. 2010 / 065152, International Publication Nos. 2011 / 153129, International Publication Nos. 2011 / 133224, International Publication Nos. 2011 / 153135, International Publication Nos. 2011 / 153136, and International Publication Nos. 2011 / 153138 (the entire contents of each are included as a whole in this specification with proper attribution).

[0022] Further examples of PDE1 inhibitors suitable for use in the methods and treatments described herein can be found in: International Publication No. 2006133261A2; U.S. Patent No. 8,273,750; U.S. Patent No. 9,000,001; U.S. Patent No. 9,624,230; International Publication No. 2009075784A1; U.S. Patent No. 8,273,751; U.S. Patent No. 8,829,008; U.S. Patent No. 9, Patent No. 403,836; International Publication No. 2014151409A1; U.S. Patent No. 9,073,936; U.S. Patent No. 9,598,426; U.S. Patent No. 9,556,186; U.S. Patent Application Publication No. 2017 / 0231994A1; International Publication No. 2016022893A1; and U.S. Patent Application Publication No. 2017 / 0226117A1 (each of which, as a whole, is part of this Specified Specification by attribution).

[0023] Further examples of PDE1 inhibitors suitable for use in the methods and treatments described herein can be found in: International Publication No. 2018007249A1; U.S. Patent Application Publication No. 2018 / 0000786; International Publication No. 2015118097A1; U.S. Patent No. 9,718,832; International Publication No. 2015091805A1; U.S. Patent No. 9,701,665; U.S. Patent Application Publication No. 2015 / 0175584A1; U.S. Patent Application Publication No. 2017 / 0267664A1; International Publication No. 2016055618A1; U.S. Patent Application Publication No. 2017 / 0298072 Publication No. A1; International Publication No. 2016170064A1; U.S. Patent Application Publication No. 2016 / 0311831A1; International Publication No. 2015150254A1; U.S. Patent Application Publication No. 2017 / 0022186A1; International Publication No. 2016174188A1; U.S. Patent Application Publication No. 2016 / 0318939A1; U.S. Patent Application Publication No. 2017 / 0291903A1; International Publication No. 2018073251A1; International Publication No. 2017178350A1; and U.S. Patent Application Publication No. 2017 / 0291901A1; each, as a whole, is incorporated herein by attribution. If any description of any document incorporated herein by attribution is found to be inconsistent or contradictory to any description in this disclosure, the description in this disclosure shall be understood to prevail.

[0024] In yet another embodiment, the present invention provides a PDE1 inhibitor for use in the therapeutic and prophylactic methods described herein, comprising enantiomers, diastereomers and racemates, in free form, salt form or prodrug form, of formula I: [ka] [In the formula, (i) R1 is H or C 1-4 It is alkyl (for example, methyl); (ii) R4 is H or C 1-4 It is an alkyl group, and R2 and R3 are independently H or C 1-4 Alkyl (For example, both R2 and R3 are methyl, or R2 is H and R3 is isopropyl), aryl, heteroaryl, (optionally hetero)arylalkoxy, or (optionally hetero)arylalkyl; or R2 is H, and R3 and R4 together form a dimethylene bridge, trimethylene bridge or tetramethylene bridge (Preferably, R3 and R4 have a cis configuration together, for example, the carbons carrying R3 and R4 have the R configuration and the S configuration respectively); (iii) R5 is a substituted heteroarylalkyl, for example substituted with haloalkyl; or R5 is bonded to one of the nitrogens of the pyrazolo moiety of formula I and has the formula A: [Chemical formula] (wherein X, Y and Z are independently N or C, and R8, R9, R 11 and R 12 are independently H or halogen (for example, Cl or F), and R 10 [[ID=2I]]is halogen, alkyl, cycloalkyl, haloalkyl (for example, trifluoromethyl), aryl (for example, phenyl), heteroaryl (for example, pyridyl (for example, pyrid-2-yl), or thiadiazolyl (for example, 1,2,3-thiadiazol-4-yl), optionally substituted with halogen), diazolyl, triazolyl, tetrazolyl, arylcarbonyl (for example, benzoyl), alkylsulfonyl (for example, methylsulfonyl), heteroarylcarbonyl, or alkoxycarbonyl; provided that when X, Y or Z is nitrogen, R8, R9 or R 10 do not exist) is the moiety represented by; (iv) R6 is H, alkyl, aryl, heteroaryl, arylalkyl (e.g., benzyl), arylamino (e.g., phenylamino), heteroarylamino, N,N-dialkylamino, N,N-diarylamino, or N-aryl-N-(arylalkyl)amino (e.g., N-phenyl-N-(1,1'-bifen-4-ylmethyl)amino); (v) n is either 0 or 1; (vi) If n is 1, then A is -C(R 13 R 14 )- and, Here, R 13 and R 14 These are, independently, H, or C 1-4 [They are alkyl, aryl, heteroaryl, (possibly hetero)arylalkoxy, or (possibly hetero)arylalkyl.] It provides that the compound is represented by [the formula shown].

[0025] In another embodiment, the present invention relates to a PDE1 inhibitor for use in the method described herein, comprising enantiomers, diastereomers and racemates, in free form, pharmaceutically acceptable salt form or prodrug form, of formula 1a: [ka] [In the formula, (i) R2 and R5 are independently H or hydroxy, and R3 and R4 together form a trimethylene bridge or a tetramethylene bridge [preferably, the carbon supporting R3 and R4 has an R configuration and an S configuration, respectively]; or R2 and R3 are each methyl, and R4 and R5 are each H; or R2, R4 and R5 are H, and R3 is isopropyl [preferably, the carbon supporting R3 has an R configuration]; (ii) R6 may be phenylamino (which may be halo-substituted or hydroxy-substituted), benzylamino (which may be halo-substituted or hydroxy-substituted), C 1-4 Alkyl or C 1-4It is an alkyl sulfide; for example, phenylamino or 4-fluorophenylamino; (iii) R 10 C 1-4 Alkyl, methylcarbonyl, hydroxyethyl, carboxylic acid, sulfonamide, phenyl (which may be halo-substituted or hydroxy-substituted), pyridyl (e.g., 6-fluoropyridi-2-yl) (which may be halo-substituted or hydroxy-substituted), or thiadiazolyl (e.g., 1,2,3-thiadiazole-4-yl); X and Y are independently C or N. It provides that.

[0026] In another embodiment, the present invention provides a PDE1 inhibitor for use in the therapeutic and prophylactic methods described herein, in free form, salt form or prodrug form, of formula II: [ka] [In the formula, (i) X is C 1-6 Alkylenes (e.g., methylene, ethylene, or propane-2-in-1-ylene); (ii) Y is a single bond, an alkynylene (e.g., -C≡C-), an arylene (e.g., phenylene), or a heteroarylene (e.g., pyridylene); (iii) Z is H, aryl (e.g., phenyl), heteroaryl (e.g., pyridyl, e.g., pyridi-2-yl), halo (e.g., F, Br, Cl), halo C 1-6 Alkyl (e.g., trifluoromethyl), -C(O)-R 1 , -N(R 2 )(R 3 ), or C which may contain at least one atom selected from the group consisting of N or O. 3-7 It is a cycloalkyl (e.g., cyclopentyl, cyclohexyl, tetrahydro-2H-pyran-4-yl, or morpholinyl); (iv) R 1 C 1-6 Alkyl, Halo C1-6 Alkyl, -OH, or -OC 1-6 It is alkyl (e.g., -OCH3); (v) R 2 and R 3 H or C 1-6 It is alkyl; (compared to) R 4 and R 5 H and C are independent of each other. 1-6 Alkyl, or one or more halos (e.g., fluorophenyl, e.g., 4-fluorophenyl), hydroxy (e.g., hydroxyphenyl, e.g., 4-hydroxyphenyl or 2-hydroxyphenyl), or C 1-6 An aryl (e.g., phenyl) which may be substituted with an alkoxy; (vii) Here, X, Y, and Z are independently one or more halos (e.g., F, Cl, or Br), C 1-6 Alkyl (e.g., methyl), halo C 1-6 It may be substituted with an alkyl group (e.g., trifluoromethyl), for example, Z may be one or more halos (e.g., 6-fluoropyrididi-2-yl, 5-fluoropyrididi-2-yl, 6-fluoropyrididi-2-yl, 3-fluoropyrididi-2-yl, 4-fluoropyrididi-2-yl, 4,6-dichloropyrididi-2-yl), halo C 1-6 Alkyl (e.g., 5-trifluoromethylpyridi-2-yl) or C 1-6 Z is either a heteroaryl, e.g., pyridyl, substituted with an alkyl (e.g., 5-methylpyridi-2-yl), or an aryl, e.g., phenyl (e.g., 4-fluorophenyl), substituted with one or more halos. It provides that the compound is represented by [the formula shown].

[0027] In yet another embodiment, the present invention provides a PDE1 inhibitor for use in the therapeutic and prophylactic methods described herein, in free or salt form, of formula III: [ka] [In the formula, (i) R1 is H or C 1-4 Alkyl (e.g., methyl or ethyl); (ii) R2 and R3 independently of H or C 1-6 Alkyl (e.g., methyl or ethyl); (iii) R4 is H or C 1-4 Alkyl (e.g., methyl or ethyl); (iv) R5 is -C(=O)-C 1-6 Alkyl (e.g., -C(=O)-CH3) and C 1-6 An aryl (e.g., phenyl) which may be substituted with one or more groups independently selected from hydroxyalkyl (e.g., 1-hydroxyethyl); (v) R6 and R7 are independently H or C 1-6 Aryl (e.g., phenyl) which may be substituted with one or more groups independently selected from alkyl (e.g., methyl or ethyl) and halogen (e.g., F or Cl), e.g., unsubstituted phenyl, or phenyl substituted with one or more halogens (e.g., F), or one or more C 1-6 Phenyl substituted with alkyl and one or more halogens, or one C 1-6 Phenyls substituted with alkyl and one halogen, such as 4-fluorophenyl, 3,4-difluorophenyl, or 4-fluoro-3-methylphenyl; (vi) n is 1, 2, 3 or 4] To provide

[0028] In yet another embodiment, the present invention provides a PDE1 inhibitor for use in the therapeutic and prophylactic methods described herein, in free or salt form, of formula IV: [ka] [In the formula, (i) R1 is C 1-4Alkyl (e.g., methyl or ethyl), or -NH(R2), where R2 is a phenyl, e.g., 4-fluorophenyl, which may be substituted with a halo (e.g., fluoro); (ii) X, Y, and Z are independently N or C; (iii) R3, R4 and R5 are independently H or C 1-4 It is alkyl (e.g., methyl); or R3 is H, and R4 and R5 together form a trimethylene bridge (preferably R4 and R5 together have a cis configuration, for example, the carbon supporting R4 and R5 has an R configuration and an S configuration, respectively). (iv) R6, R7 and R8 independently: H, C 1-4 Alkyl (e.g., methyl), Pyridi-2-yl substituted with hydroxyl, or -S(O)2-NH2 and; (v) However, if X, Y and / or Z are N, then R6, R7 and / or R8 are not present, respectively; and if X, Y and Z are all C, then at least one of R6, R7 or R8 is -S(O)2-NH2 or pyridi-2-yl substituted with hydroxyl. It provides that.

[0029] In another embodiment, the present invention provides a PDE1 inhibitor for use in the method described herein, in free or salt form, of formula V: [ka] [In the formula, (i) R1 is -NH(R4), where R4 is a phenyl which may be substituted with a halo (e.g., fluoro), e.g., 4-fluorophenyl; (ii) R2 is H or C 1-6 Alkyl (e.g., methyl, isobutyl, or neopentyl); (iii) R3 is -SO2NH2 or -COOH] It provides that.

[0030] In another embodiment, the present invention provides a PDE1 inhibitor for use in the method described herein, in free form or salt form, of formula VI: [ka] [In the formula, (i) R1 is -NH(R4), where R4 is a phenyl which may be substituted with a halo (e.g., fluoro), such as 4-fluorophenyl; (ii) R2 is H or C 1-6 Alkyl (e.g., methyl or ethyl); (iii) R3 is H, halogen (e.g., bromo), C 1-6 [These are alkyl (e.g., methyl), aryl (e.g., 4-fluorophenyl) which may be substituted with a halogen, heteroaryl (e.g., 6-fluoropyrididi-2-yl or pyridi-2-yl) which may be substituted with a halogen, or acyl (e.g., acetyl)] It provides that.

[0031] In one embodiment, the present disclosure provides an administration of a PDE1 inhibitor (e.g., a compound represented by formula I, Ia, II, III, IV, V, and / or VI) for use in the method described herein, wherein the inhibitor is one of the following compounds: [ka] [ka] [ka] [ka] It provides administration

[0032] In one embodiment, the present invention relates to the administration of a PDE1 inhibitor for the treatment or prevention of inflammation or inflammation-related disease or disorder, wherein the inhibitor is a compound of the following forms, either in free form or in pharmaceutically acceptable salt form: [ka] Therefore, it provides administration.

[0033] In yet another embodiment, the present invention relates to the administration of a PDE1 inhibitor for the treatment or prevention of inflammation or inflammation-related disease or disorder, wherein the inhibitor is a compound in free form or pharmaceutically acceptable salt form: [ka] Therefore, it provides administration.

[0034] In yet another embodiment, the present invention relates to the administration of a PDE1 inhibitor for the treatment or prevention of inflammation or inflammation-related disease or disorder, wherein the inhibitor is a compound in free form or pharmaceutically acceptable salt form: [ka] Therefore, it provides administration.

[0035] In yet another embodiment, the present invention relates to the administration of a PDE1 inhibitor for the treatment or prevention of inflammation or inflammation-related disease or disorder, wherein the inhibitor is a compound in free form or pharmaceutically acceptable salt form: [ka] Therefore, it provides administration.

[0036] In one embodiment, a selective PDE1 inhibitor represented by any of the above formulas (e.g., formulas I, Ia, II, III, IV, V, and / or VI) is a compound that inhibits phosphodiesterase-mediated (e.g., PDE1-mediated, particularly PDE1B-mediated) hydrolysis of cGMP, for example, preferred compounds in free or salt form exhibiting an IC50 of less than 1 μM, preferably less than 500 nM, preferably less than 50 nM, and preferably less than 5 nM in an immobilized metal affinity particle reagent PDE assay. 50 It holds.

[0037] Unless otherwise specified or evident from the context, the following terms used herein have the following meanings:

[0038] As used herein, "alkyl" is a saturated or unsaturated hydrocarbon moiety, preferably saturated, preferably having 1 to 6 carbon atoms, and may be linear or branched, and may be monosubstituted, disubstituted or trisubstituted with, for example, halogens (e.g., chloro or fluoro), hydroxy, or carboxy.

[0039] As used herein, "cycloalkyl" is a saturated or unsaturated non-aromatic hydrocarbon moiety, preferably saturated, and preferably containing 3 to 9 carbon atoms, at least some of which form a non-aromatic monocyclic, bicyclic, or bridging cyclic structure, which may be substituted with, for example, halogens (e.g., chloro or fluoro), hydroxy, or carboxy. If the cycloalkyl may contain one or more atoms selected from N and O and / or S, then the cycloalkyl may also be a heterocycloalkyl.

[0040] Unless otherwise specified, "heterocycloalkyl" is a saturated or unsaturated non-aromatic hydrocarbon moiety, preferably saturated, and preferably containing 3 to 9 carbon atoms, where at least some of the carbon atoms form a non-aromatic monocyclic, bicyclic, or bridging cyclic structure, where at least one carbon atom is replaced by N, O, or S, and the heterocycloalkyl may be substituted with, for example, a halogen (e.g., chloro or fluoro), hydroxy, or carboxy.

[0041] As used herein, "aryl" is a monocyclic or bicyclic aromatic hydrocarbon, preferably phenyl, which may be substituted with, for example, alkyl (e.g., methyl), halogen (e.g., chloro or fluoro), haloalkyl (e.g., trifluoromethyl), hydroxy, carboxy, or further aryl or heteroaryl (e.g., biphenyl or pyridylphenyl).

[0042] As used herein, "heteroaryl" is an aromatic moiety in which one or more atoms constituting the aromatic ring are sulfur or nitrogen rather than carbon, for example pyridyl or thiadiazolyl, and may be substituted with alkyl, halogen, haloalkyl, hydroxy or carboxy, for example.

[0043] The compounds of the present invention, for example, the substituted 7,8-dihydroimidazo[1,2-a]pyrazolo[4,3-e]pyrimidine-4-one compounds and 7,8,9-trihydro-[1H or 2H]-pyrimido[1,2-a]pyrazolo[4,3-e]pyrimidine-4(5H)-one compounds, for example, the compounds represented by formulas I, Ia, II, III, IV, V, and / or VI, may exist in free form or as salts, for example, as acid addition salts. In this specification, unless otherwise specified, terms such as “compounds of the present invention” should be understood to encompass compounds in all forms, for example, free form or acid addition salt form, or, if the compound contains an acidic substituent, the base addition salt form. The compounds of the present invention are intended for pharmaceutical use, and therefore, pharmaceutically acceptable salts are preferred. Salts unsuitable for medicinal use may be useful, for example, for the isolation or purification of the free compounds of the present invention or their pharmaceutically acceptable salts, and are therefore included as well.

[0044] The compounds of the present invention may, in some cases, also exist in prodrug form. A prodrug form is a compound that is converted into the compound of the present invention in the body. For example, if the compound of the present invention contains a hydroxy substituent or a carboxy substituent, these substituents may form a physiologically hydrolyzable and acceptable ester. As used herein, “physiologically hydrolyzable and acceptable ester” means an ester of the compound of the present invention that is hydrolyzable under physiological conditions and produces a physiologically acceptable acid (in the case of the compound of the present invention having a hydroxy substituent) or alcohol (in the case of the compound of the present invention having a carboxy substituent) at the dose administered. Therefore, if the compound of the present invention contains a hydroxyl group, for example, compound -OH, then an acyl ester prodrug of such a compound, i.e., compound -OC(O)-C, may be formed. 1-4 Alkyl compounds undergo hydrolysis in the body, yielding, on the one hand, physiologically hydrolyzable alcohols (compounds -OH), and on the other hand, acids (e.g., HOC(O)-C). 1-4Alkyl compounds can be formed. Alternatively, if the compound of the present invention contains a carboxylic acid, for example, compound -C(O)OH, then an acid ester prodrug of such a compound, i.e., compound -C(O)OC 1-4 Alkyl compounds undergo hydrolysis to form compounds -C(O)OH and HO-C 1-4 Alkyl compounds can be formed. Therefore, as is to be understood, this term encompasses the conventional pharmacoprodrug forms.

[0045] In another embodiment, the present invention also provides a pharmaceutical composition for use as an anti-inflammatory agent, comprising a mixture of the compound of the present invention in free form or in pharmaceutically acceptable salt form with a pharmaceutically acceptable carrier.

[0046] The compounds of the present invention may, in some cases, also exist in prodrug form. A prodrug form is a compound that is converted into the compound of the present invention in the body. For example, if the compound of the present invention contains a hydroxy substituent or a carboxy substituent, these substituents may form a physiologically hydrolyzable and acceptable ester. As used herein, “physiologically hydrolyzable and acceptable ester” means an ester of the compound of the present invention that is hydrolyzable under physiological conditions and produces a physiologically acceptable acid (in the case of the compound of the present invention having a hydroxy substituent) or alcohol (in the case of the compound of the present invention having a carboxy substituent) at the dose administered. Therefore, if the compound of the present invention contains a hydroxyl group, for example, compound -OH, then an acyl ester prodrug of such a compound, i.e., compound -OC(O)-C, may be formed. 1-4 Alkyl compounds undergo hydrolysis in the body, yielding, on the one hand, physiologically hydrolyzable alcohols (compounds -OH), and on the other hand, acids (e.g., HOC(O)-C). 1-4 Alkyl compounds can be formed. Alternatively, if the compound of the present invention contains a carboxylic acid, for example, compound -C(O)OH, then an acid ester prodrug of such a compound, i.e., compound -C(O)OC 1-4 Alkyl compounds undergo hydrolysis to form compounds -C(O)OH and HO-C1-4 Alkyl compounds can be formed. Therefore, as is to be understood, this term encompasses the conventional pharmacoprodrug forms.

[0047] In another embodiment, the present invention also provides a pharmaceutical composition for use as an anti-inflammatory agent, comprising a compound of the present invention in free form, pharmaceutically acceptable salt form, or prodrug form, mixed with a pharmaceutically acceptable carrier.

[0048] Method for producing the compound of the present invention The compounds of the present invention and their pharmaceutically acceptable salts may be produced using the methods described and illustrated herein, and similarly, and by methods known in the chemical art. Such methods include, but are not limited to, the following. If the starting materials for these methods are not commercially available, they may be produced by procedures selected from the chemical art using techniques similar to or similar to those used in the synthesis of known compounds.

[0049] Various starting materials and / or compounds of the present invention are published in U.S. Patent Publication No. 2008-0188492A1, U.S. Patent Publication No. 2010-0173878A1, U.S. Patent Publication No. 2010-0273754A1, U.S. Patent Publication No. 2010-0273753A1, International Publication No. 2010 / 065153, International Publication No. 2010 / 065151, International Publication No. 2010 / 065151, International Publication No. 2010 / 065149, International Publication It may be manufactured using the methods described in Publication No. 2010 / 065147, International Publication No. 2010 / 065152, International Publication No. 2011 / 153129, International Publication No. 2011 / 133224, International Publication No. 2011 / 153135, International Publication No. 2011 / 153136, International Publication No. 2011 / 153138, and U.S. Patent No. 9,073,936 (each of which, with due attribution, constitutes part of this Specified Specified Specified Specified as a whole).

[0050] The compounds of the present invention encompass their enantiomers, diastereoisomers, and racemates, as well as their polymorphs, hydrates, solvates, and complexes. Some individual compounds within the scope of the present invention may contain a double bond. The expression "double bond" in the present invention means that both the E and Z isomers of the double bond are included. In addition, some compounds within the scope of the present invention may contain one or more chiral centers. The present invention encompasses the use of any one or more optically pure stereoisomers and combinations of stereoisomers.

[0051] The compounds of the present invention are also intended to include their stable and unstable isotopes. Stable isotopes are non-radioactive isotopes that contain one additional neutron compared to the abundant nuclide (i.e., element) of the same type. The activity of compounds containing such isotopes is preserved, and such compounds are also expected to be useful for measuring the pharmacokinetics of non-isotope analogs. For example, a hydrogen atom at a certain position in the compounds of the present invention can be replaced with deuterium (a stable isotope that is non-radioactive). Examples of known stable isotopes include deuterium, 13 C, 15 N, 18 O is one example, but it is not limited to these. Alternatively, unstable isotopes that contain more neutrons compared to the same type of nuclide (i.e., element) can be considered, for example, 123 I, 131 I, 125 I, 11 C, 18 F can be replaced by the corresponding abundant species of I, C, and F. Another example of a useful isotope of the compound of the present invention is: 11 These are 13C isotopes. These radioactive isotopes are useful for radioimaging and / or pharmacokinetic studies of the compounds of the present invention.

[0052] Melting points are uncorrected, and (dec) indicates decomposition. Temperatures are given in degrees Celsius (°C); unless otherwise specified, operations are performed at room temperature or ambient temperature, i.e., in the range of 18–25°C. Chromatography refers to flash silica gel chromatography; thin-layer chromatography (TLC) is performed on silica gel plates. NMR data are the delta values ​​of major characteristic protons, given in parts per million (ppm) relative to tetramethylsilane (TMS) as an internal standard. Conventional abbreviations for signal shape are used. Coupling constants (J) are given in Hz. For mass spectra (MS), the lowest mass principal ion is reported for molecules where isotopic fission yields multiple mass spectral peaks. Solvent mixture composition is given as volume percentage or volume ratio. If the NMR spectrum is complex, only characteristic signals are reported.

[0053] Method of using the compound of the present invention The compounds of the present invention are useful for the treatment of inflammatory diseases or conditions, particularly inflammatory diseases or conditions. Accordingly, the administration or use of preferred PDE1 inhibitors described herein, such as the above-mentioned PDE1 inhibitors, for example, compounds represented by formulas I, Ia, II, III, IV, V and / or VI, provides a means to modulate inflammation (e.g., to prevent, alleviate, and / or promote recovery from inflammation and inflammation-related diseases or disorders), and in some embodiments, provides the treatment of a variety of inflammatory diseases and disorders.

[0054] In one embodiment, the present invention provides a method for promoting the resolution of inflammation, comprising administering an effective amount of a phosphodiesterase type I specific inhibitor (PDE1) to a patient in need of such promotion (Method 1), for example:

[0055] For example, in one embodiment, the present invention provides a method for treating or preventing inflammation or inflammation-related diseases (Method 1), for example, the following method, which includes administering an effective amount of a specific inhibitor of type I phosphodiesterase (PDE1) to a patient in need:

[0056] 1.1. The patient suffers from a disease or disorder mediated by inflammation and / or macrophage activation, Method 1.

[0057] 1.2. Method 1 or 1.1, which includes promoting the resolution of inflammation by promoting the activation of M2 macrophages.

[0058] 1.3. Diseases or conditions to be treated include: bacterial infections (e.g., infections with Salmonella typhi, Salmonella typhimurium, Listeria monocytogenes, Mycobacterium tuberculosis, Mycobacterium ulcerans, and Mycobacterium avium); viral infections (e.g., African Swine Fever Virus, Classical Swine Fever Virus, Dengue Virus, Foot and Mouth Disease Virus, Human Immunodeficiency Virus (HIV) (e.g., HIV1), and Influenza A virus). Viruses, Porcine Circovirus-2, Porcine Reproductive and Respiratory Syndrome Virus, Porcine Pseudorabies Virus, Respiratory Syncytial Virus, Severe Acute Respiratory Syndrome Coronavirus, West Nile Virus, Viral Hepatitis (e.g., Hepatitis A, Hepatitis B, Hepatitis C); Parasitic Infections (e.g., Taenia crassiceps, Toxoplasma gondii, Leishmania infantum, Schistosoma mansoni infections); Atopic Dermatitis; Pneumonia; Cardiovascular Diseases, e.g., Atherosclerosis;Method 1 or 1.1, selected from obesity and insulin resistance; asthma; pulmonary fibrosis; cardiac obstructive pulmonary disease (COPD); neuropathic pain; stroke; diabetes mellitus; sepsis; non-alcoholic steatohepatitis (NASH); autoimmune hepatitis; systemic lupus erythematosus (SLE); wound healing; pleurisy; peritonitis; and cystic fibrosis.

[0059] 1.4. Any of the above methods, where the disease or condition to be treated is a bacterial infection.

[0060] 1.5. Any of the above methods, in which the disease or condition to be treated is an infection of Salmonella typhi, Salmonella typhimurium, Listeria monocytogenes, Mycobacterium tuberculosis, Mycobacterium ulcerans, or Mycobacterium avium.

[0061] 1.6. If the disease or condition to be treated is a viral infection, follow methods 1-1.3.

[0062] 1.7. Viral infections include African Swine Fever Virus, Classical Swine Fever Virus, Dengue Virus, Foot and Mouth Disease Virus, Human Immunodeficiency Virus (HIV) (e.g., HIV1), Influenza A Virus, Porcine Circovirus-2, Porcine Reproductive and Respiratory Syndrome Virus, Porcine Pseudorabies Virus, Respiratory Syncytial Virus, Severe Acute Respiratory Syndrome Coronavirus, and West Nile Virus. Method 1.6, which is a virus, or viral hepatitis (e.g., hepatitis A, hepatitis B, hepatitis C).

[0063] 1.8. If the disease or condition to be treated is a parasitic infection, follow methods 1-1.3.

[0064] 1.9. If the parasitic infection is an infection with Taenia crassiceps, Toxoplasma gondii, Leishmania infantum, or Schistosoma mansoni, Method 1.8.

[0065] 1.10. If the disease or condition to be treated is atopic dermatitis; pneumonia; cardiovascular disease, e.g., atherosclerosis; obesity and insulin resistance; asthma; pulmonary fibrosis; cardiac obstructive pulmonary disease (COPD); neuropathic pain; stroke; diabetes; sepsis; non-alcoholic steatohepatitis (NASH); autoimmune hepatitis; systemic lupus erythematosus (SLE); wound healing; pleurisy; peritonitis; and cystic fibrosis, then methods 1-1.3.

[0066] 1.11. Methods 1-1.3 or 1.10, where the disease or condition to be treated is non-alcoholic steatohepatitis (NASH); autoimmune hepatitis; systemic lupus erythematosus (SLE); wound healing; pleurisy; peritonitis; and cystic fibrosis.

[0067] 1.12. Any of the above methods, comprising administering an effective amount of the PDE1 inhibitor of the present invention (e.g., a PDE1 inhibitor represented by formulas I, Ia, II, III, IV, V and / or VI as described herein) to a patient in need, in an amount effective to (i) reduce or inhibit the activation of M1 macrophages, and / or (ii) reduce the levels of one or more pro-inflammatory cytokines (e.g., IL1β, TNFα, IL6 and Ccl2, or a combination thereof).

[0068] 1.13. Any of the above methods, comprising administering an effective amount of the PDE1 inhibitor of the present invention (for example, a PDE1 inhibitor represented by formulas I, Ia, II, III, IV, V and / or VI as described herein) to a patient in need, in an amount effective to (i) promote the activation of M2 macrophages, and / or (ii) promote the production of anti-inflammatory cytokines (e.g., IL-10).

[0069] 1.14. Any of the above methods, comprising administering an effective amount of the PDE1 inhibitor of the present invention (for example, a PDE1 inhibitor represented by formulas I, Ia, II, III, IV, V and / or VI as described herein) to a patient in need in an amount effective to decrease the level of M1 phenotype macrophages and / or increase the level of M2 phenotype macrophages.

[0070] 1.15. Any of the above methods, wherein the PDE1 inhibitor is a compound represented by formula I, Ia, II, III, IV, V and / or VI.

[0071] 1.16. Inflammation is associated with increased expression and / or activation of macrophages (e.g., M1 macrophages) in any of the above ways.

[0072] 1.17. Any of the above methods, wherein a PDE1 inhibitor slows down or inhibits the expression and / or activity of pro-inflammatory cytokines, selected from the group consisting of, for example, IL1B, IL-6, TNFα, Ccl2, nitric oxide (NO), and reactive oxygen species (ROS).

[0073] 1.18. Any of the above methods, in which a PDE1 inhibitor is administered in combination with a PDE4 inhibitor (e.g., lolipram).

[0074] 1.19. The patient shows elevated levels of pro-inflammatory cytokines (e.g., IL1B, IL6, TNFα, Ccl2) by any of the above methods.

[0075] 1.20. When the "PDE1 inhibitor" has an IC of less than 1 μM, preferably less than 750 nM, more preferably less than 500 nM, and more preferably less than 50 nM in, for example, an immobilized metal affinity particle reagent PDE assay, 50 The above method represents a compound that selectively inhibits phosphodiesterase-mediated (e.g., PDE1-mediated, particularly PDE1B-mediated) hydrolysis of cGMP.

[0076] 1.21. PDE1 inhibitors have an IC50 of less than 10 nM. 50 This inhibits the activity of PDE1 (for example, bovine PDE1 in the assay described in Example 1), and for example, the PDE1 inhibitor does not inhibit the activity of PDEs other than PDE1, and for example, it inhibits IC50 at least 1000 times compared to PDEs other than PDE1. 50 Having any of the above methods.

[0077] 1.22. PDE1 inhibitors, in free form or pharmaceutically acceptable form, as follows: [ka] One of the above methods.

[0078] 1.23. PDE1 inhibitors, in free form or pharmaceutically acceptable form, as follows: [ka] One of the above methods.

[0079] 1.24. PDE1 inhibitors, in free form or pharmaceutically acceptable form, as follows: [ka] One of the above methods.

[0080] 1.25. PDE1 inhibitors, in free form or pharmaceutically acceptable form, as follows: [ka] One of the above methods.

[0081] 1.26. Any of the above methods, in which the patient has elevated levels of one or more pro-inflammatory cytokines (e.g., selected from IL1β, TNFα, Ccl2, IL-6, and combinations thereof).

[0082] 1.27. Any of the above methods, wherein the patient has a reduced level of one or more anti-inflammatory cytokines (e.g., IL-10).

[0083] 1.28. Any of the above methods, wherein the patient has elevated levels of M1 phenotype macrophages compared to M2 phenotype macrophages.

[0084] 1.29. The patient is further administered one or more of the following: antibiotics, antivirals, corticosteroids, or NSAIDs, in any of the manner described above.

[0085] For example, in one embodiment, the present invention provides a method (Method 2) for promoting macrophage activation to an M2 activated state, comprising administering an effective amount of a phosphodiesterase type I specific inhibitor (PDE1) to a patient suffering from inflammation or an inflammation-related disease or condition (e.g., macrophage-mediated inflammation), for example:

[0086] 2.1 Diseases or conditions to be treated include bacterial infections (e.g., infections with Salmonella typhi, Salmonella typhimurium, Listeria monocytogenes, Mycobacterium tuberculosis, Mycobacterium ulcerans, and Mycobacterium avium); viral infections (e.g., African Swine Fever Virus, Classical Swine Fever Virus, Dengue Virus, Foot and Mouth Disease Virus, Human Immunodeficiency Virus (HIV) (e.g., HIV1), and Influenza A virus). Viruses, Porcine Circovirus-2, Porcine Reproductive and Respiratory Syndrome Virus, Porcine Pseudorabies Virus, Respiratory Syncytial Virus, Severe Acute Respiratory Syndrome Coronavirus, West Nile Virus, Viral Hepatitis (e.g., Hepatitis A, Hepatitis B, Hepatitis C); Parasitic Infections (e.g., Taenia crassiceps, Toxoplasma gondii, Leishmania infantum, Schistosoma mansoni infections); Atopic Dermatitis; Pneumonia; Cardiovascular Diseases, e.g., Atherosclerosis;Method 2, selected from obesity and insulin resistance; asthma; pulmonary fibrosis; cardiac obstructive pulmonary disease (COPD); neuropathic pain; stroke; diabetes; sepsis; non-alcoholic steatohepatitis (NASH); autoimmune hepatitis; systemic lupus erythematosus (SLE); wound healing; pleurisy; peritonitis; and cystic fibrosis.

[0087] 2.2 Any of the above methods, where the disease or condition to be treated is a bacterial infection.

[0088] 2.3 Any of the above methods, in which the disease or condition to be treated is an infection of Salmonella typhi, Salmonella typhimurium, Listeria monocytogenes, Mycobacterium tuberculosis, Mycobacterium ulcerans, or Mycobacterium avium.

[0089] 2.4 If the disease or condition to be treated is a viral infection, Method 2 or 2.1.

[0090] 2.5 Viral infections include African Swine Fever Virus, Classical Swine Fever Virus, Dengue Virus, Foot and Mouth Disease Virus, Human Immunodeficiency Virus (HIV) (e.g., HIV1), Influenza A Virus, Porcine Circovirus-2, Porcine Reproductive and Respiratory Syndrome Virus, Porcine Pseudorabies Virus, Respiratory Syncytial Virus, Severe Acute Respiratory Syndrome Coronavirus, and West Nile Virus. Method 2 or 2.4, which is a virus, or viral hepatitis (e.g., hepatitis A, hepatitis B, hepatitis C).

[0091] 2.6 If the disease or condition to be treated is a parasitic infection, Method 2 or 2.1.

[0092] 2.7 The parasitic infection is an infection with Taenia crassiceps, Toxoplasma gondii, Leishmania infantum, or Schistosoma mansoni, Method 2.7.

[0093] 2.8 Method 2 or 2.1, where the disease or condition to be treated is atopic dermatitis; pneumonia; cardiovascular disease, e.g., atherosclerosis; obesity and insulin resistance; asthma; pulmonary fibrosis; cardiac obstructive pulmonary disease (COPD); neuropathic pain; stroke; diabetes mellitus; sepsis; non-alcoholic steatohepatitis (NASH); autoimmune hepatitis; systemic lupus erythematosus (SLE); wound healing; pleurisy; peritonitis; and cystic fibrosis.

[0094] 2.9 Method 2 or 2.8, where the disease or condition to be treated is non-alcoholic steatohepatitis (NASH); autoimmune hepatitis; systemic lupus erythematosus (SLE); wound healing; pleurisy; peritonitis; and cystic fibrosis.

[0095] 2.10 Any of the above methods, comprising administering an effective amount of the PDE1 inhibitor of the present invention (for example, a PDE1 inhibitor represented by formulas I, Ia, II, III, IV, V and / or VI as described herein) to a patient in need, in an amount effective to (i) reduce or inhibit the activation of M1 macrophages, and / or (ii) reduce the levels of one or more pro-inflammatory cytokines (e.g., IL1β, TNFα, IL6 and Ccl2, or a combination thereof).

[0096] 2.11 Any of the above methods, comprising administering an effective amount of the PDE1 inhibitor of the present invention (for example, a PDE1 inhibitor represented by formulas I, Ia, II, III, IV, V and / or VI as described herein) to a patient in need, in an amount effective to (i) promote the activation of M2 macrophages, and / or (ii) promote the production of anti-inflammatory cytokines (e.g., IL-10).

[0097] 2.12 Any of the above methods, comprising administering an effective amount of the PDE1 inhibitor of the present invention (for example, a PDE1 inhibitor represented by formulas I, Ia, II, III, IV, V and / or VI as described herein) to a patient in need in an amount effective to reduce the level of M1 phenotype macrophages and / or increase the level of M2 phenotype macrophages.

[0098] 2.13 Any of the above methods, wherein the PDE1 inhibitor is a compound represented by formula I, Ia, II, III, IV, V and / or VI.

[0099] 2.14 Inflammation is associated with any of the above methods, specifically with increased expression and / or activation of macrophages (e.g., M1 macrophages).

[0100] 2.15 Any of the above methods, wherein a PDE1 inhibitor slows down or inhibits the expression and / or activity of pro-inflammatory cytokines, selected from the group consisting of, for example, IL1B, IL-6, TNFα, Ccl2, nitric oxide (NO), and reactive oxygen species (ROS).

[0101] 2.16 Any of the above methods, in which a PDE1 inhibitor is administered in combination with a PDE4 inhibitor (e.g., lolipram).

[0102] 2.17 The patient shows elevated levels of pro-inflammatory cytokines (e.g., IL1B, IL6, TNFα, Ccl2) by any of the above methods.

[0103] 2.18 If the "PDE1 inhibitor" has an IC of less than 1 μM, preferably less than 750 nM, more preferably less than 500 nM, and more preferably less than 50 nM in, for example, an immobilized metal affinity particle reagent PDE assay, 50 The above method represents a compound that selectively inhibits phosphodiesterase-mediated (e.g., PDE1-mediated, particularly PDE1B-mediated) hydrolysis of cGMP.

[0104] 2.19 PDE1 inhibitors have an IC50 of less than 10 nM 50 This inhibits the activity of PDE1 (for example, bovine PDE1 in the assay described in Example 1), and for example, the PDE1 inhibitor does not inhibit the activity of PDEs other than PDE1, and for example, it inhibits IC50 at least 1000 times compared to PDEs other than PDE1. 50 Having any of the above methods.

[0105] 2.20 PDE1 inhibitors, either in free form or pharmaceutically acceptable salt form, as follows: [ka] One of the above methods.

[0106] 2.21 PDE1 inhibitors, either in free form or pharmaceutically acceptable salt form, as follows: [ka] One of the above methods.

[0107] 2.22 PDE1 inhibitors, in free form or pharmaceutically acceptable form, include the following: [ka] One of the above methods.

[0108] 2.23 PDE1 inhibitors, in free form or pharmaceutically acceptable form, as follows: [ka] One of the above methods.

[0109] 2.24 Any of the above methods, wherein the patient has elevated levels of one or more pro-inflammatory cytokines (e.g., selected from IL1β, TNFα, Ccl2, IL-6, and combinations thereof).

[0110] 2.25 Any of the above methods, wherein the patient has a reduced level of one or more anti-inflammatory cytokines (e.g., IL-10).

[0111] 2.26 Any of the above methods, wherein the patient has elevated levels of M1 phenotype macrophages compared to M2 phenotype macrophages.

[0112] 2.27 The patient is further administered one or more of the following: antibiotics, antivirals, corticosteroids, or NSAIDs, in any of the manner described above.

[0113] The present invention further provides the use of any of the PDE1 inhibitors, such as compounds represented by formulas I, Ia, II, III, IV, V and / or VI, in the manufacture of a pharmaceutical for use in any of the methods from Method 1 onward.

[0114] The present invention further provides PDE1 inhibitors, for example, any of the compounds represented by formulas I, Ia, II, III, IV, V and / or VI, for use in any of the methods from Method 1 onward.

[0115] The present invention further provides a pharmaceutical composition for use in any of the methods from Method 1 onward, comprising a PDE1 inhibitor, for example, any of formulas I, Ia, II, III, IV, V, and / or VI.

[0116] The phrase "compounds of the present invention" or "PDE1 inhibitors of the present invention" or similar terms encompass any or all of the compounds described herein, for example, the compounds represented by formulas I, Ia, II, III, IV, V and / or VI.

[0117] The terms “treatment” and “to treat” should be appropriately understood to include the prevention and treatment or remission of the symptoms of a disease, as well as the treatment of the underlying cause of the disease.

[0118] Regarding treatment methods, the term “effective dose” is intended to encompass a therapeutically effective dose for treating or alleviating a particular disease or disorder and / or its symptoms, and / or for reducing inflammatory cytokines, e.g., those produced by macrophages, and / or for reducing M1 macrophage activation, and / or for increasing anti-inflammatory cytokines, e.g., those produced by macrophages, and / or for enhancing M2 macrophage activation.

[0119] The term "patient" can refer to a human or a non-human (i.e., animal) patient. In certain embodiments, the invention encompasses both humans and non-human animals. In other embodiments, the invention encompasses non-human animals. In other embodiments, it encompasses humans.

[0120] The term "including" as used in this disclosure is intended to be open-ended and does not exclude any further elements or method steps not described herein.

[0121] The compounds of the present invention, for example, in free form or pharmaceutically acceptable salt form, of formulas I, Ia, II, III, IV, V and / or VI above, may be used as single therapeutic agents, but may also be used in combination with other active agents or for co-administration.

[0122] For example, in one embodiment, the compounds of the present invention, e.g., in free form or pharmaceutically acceptable salt form, of formulas I, Ia, II, III, IV, V and / or VI above, can be administered in combination with other active agents, e.g., one or more antidepressants, e.g., one or more compounds in free form or pharmaceutically acceptable salt form selected from, e.g., selective serotonin reuptake inhibitors (SSRIs), c) serotonin-norepinephrine reuptake inhibitors (SNRIs), a) tricyclic antidepressants (TCAs), and atypical antipsychotics (e.g., they can be administered sequentially, simultaneously, or within 24 hours).

[0123] The dosage used when carrying out the present invention will, of course, vary depending on the specific disease or condition to be treated, the specific compound of the present invention used, the mode of administration, and the desired treatment. The compounds of the present invention can be administered by appropriate routes, including orally, parenterally, transdermally, or by inhalation, but are preferably administered orally. Generally, satisfactory effects for the treatment of the above-mentioned diseases, for example, have been shown to be obtained with oral administration at doses of approximately 0.01 to 2.0 mg / kg. In larger mammals, such as humans, the indicated daily dose for oral administration will accordingly range from approximately 0.75 to 150 mg (depending on the drug administered and the condition being treated, for example, in the case of compound 214, 0.5 to 25 mg per day for the treatment of inflammatory conditions, e.g., 1 to 10 mg, e.g., in the form of a monophosphate), and may be conveniently administered once daily, or in 2 to 4 divided doses, or in a sustained-release form. Therefore, a unit dosage form for oral administration may contain, for example, about 0.2 to 75 or 150 mg of the compound of the present invention, for example, about 0.2 or 2.0 to 50, 75 or 100 mg (e.g., 1, 2.5, 5, 10 or 20 mg) together with a pharmaceutically acceptable diluent or carrier for that purpose.

[0124] Pharmaceutical compositions containing the compounds of the present invention can be prepared using conventional diluents or excipients and techniques known in the Galenic art. Therefore, oral administration forms include tablets, capsules, liquids, and suspensions. [Examples]

[0125] Example 1: Evaluation of peripheral inflammation using a mouse zymosan pleurisy model To induce sterile inflammation, zymosan is injected into the pleural cavity of mice. Leukocyte, neutrophil, and macrophage infiltration is monitored on days 3 and 7 post-injection. Detection of various cell types is performed according to the gating strategy outlined in Table 1 below.

[0126] [Table 1]

[0127] In this model, injection of zymosan mobilizes various waves of leukocytes, which are observed and recorded. The amount of exudate increases to a maximum over 24 hours, neutrophils increase within 4 hours and reach a maximum at 48 hours. Lymphocytes of the adaptive immune system enter later, at 3 days, and are signaled by antigen-presenting macrophages. The demise phase is well demonstrated in this model and involves a decrease in total macrophage number and a transition to the M2 phenotype.

[0128] In these studies, subjects were administered compounds 1 and 2, and the effects of the compounds on the infiltration of leukocytes, neutrophils, and macrophages were observed.

[0129] [ka]

[0130] As shown in the attached Figures 1–9, subjects treated with compound 1 or 2 were observed to show enhanced inflammation resolution by promoting a shift from M1 to M2 macrophages. This data indicates that in treated specimens, inflammation attributable to M1 macrophages was consistently reduced, while M2 activation was promoted. As shown in Figure 1, intraperitoneal injection of zymosan 1 mgi.p. resulted in significant total CD45+ leukocyte infiltration. This increase in total leukocyte count resulted in a general increase in total macrophage count at days 3 and 7 after zymosan injection in disease-only animals compared to untreated animals (Figure 2A). The macrophage rate based on total leukocyte count (Figure 2B) decreased slightly between days 3 and 7.

[0131] Neutrophil counts decreased significantly on day 7 in disease-only animals and vehicle test animals, but not as dramatically in animals administered compound 1 (Figure 3A). These results are reflected in Figure 3B, which shows that the overall ratio of neutrophils to CD45+ leukocytes was significantly reduced in all subjects.

[0132] To further evaluate CD38 and Egr2 expression on macrophages, the total number of CD38+ and Egr2+ macrophages was analyzed. The total number of CD38+ macrophages increased in the diseased and vehicle day 3 groups, but decreased in animals treated with compound 1 on day 3 (Figure 5A). The number of Egr2+ macrophages decreased in all groups on day 3 (Figure 5B). The mean fluorescence intensity (MFI) of both CD38 and Egr2 was also analyzed in macrophages in Figures 6A and 6B. MFI provides a numerical value regarding the relative expression of a given marker in a cell population. CD38+ MFI increased in all groups on day 3, was lowest in the group treated with compound 1, and decreased in all groups on day 7. On the other hand, Egr2+ MFI decreased in all groups on day 3 and day 7 compared to the untreated group.

[0133] Overall, the results showed a tendency for the number of CD38+ cells to decrease and the number and proportion of Egr2+ cells to increase, indicating an increased phase of inflammatory injury resolution on day 7. Furthermore, as shown in Figure 7, animals treated with compound 1 also tended to show lower levels of the inflammatory biomarker (MCP-1 / CCL2) on days 3 and 7 compared to the control group.

[0134] Similar tests were conducted with compound 2, and the results are shown in Figures 8 and 9. Treatment with 2 mg / kg of dexamethasone did not significantly affect the number of CD38+ or Erg2+ macrophage populations on day 3 or day 7. However, treatment with 3 mg / kg of compound 2 resulted in a significant decrease in CD38+ macrophages and a rapid and significant increase in Erg2+ macrophages on day 7. Further tests were conducted according to the same method. Mice were intraperitoneally injected with 1 mg of zymosan, followed by intraperitoneal injection of 10 mg / kg of compound 1. Macrophage levels were recorded at the time of injection, and then at 4, 8, 16, 24, 48, and 72 hours after injection. The results are summarized in Figures 11A, 11B, 12A, and 12B. As shown in Figures 11A and 11B, treatment with compound 1 resulted in a decrease in M1 macrophage levels at all observed time points, with a significant difference observed in CD80+ macrophages on day 7. Correspondingly, Arg1+ M2 macrophages increased at hour 4, and CD206+ M2 macrophages significantly increased compared to controls on days 2 and 3.

[0135] Example 2: Effect of PDE1 inhibitor on microglial chemotaxis assay BV2 cells were added to the upper chamber of a 5 μm pore Transwell 96-well plate above a reservoir containing 100 μM ADP and incubated at 37°C and 5% CO2 for 4 hours. After incubation, cells were harvested in a pre-warmed cell detachment solution for 30 minutes under the same incubation conditions. 75 μl of this cell detachment solution was mixed with 75 μl of culture medium in a new 96-well plate compatible with a fluorescence reader. The number of cells in the lower chamber was determined by adding CyQuant® GR dye and reading with an Envision fluorescence reader at 480 nm EX / 520 nm EM. CyQuant® GR dye exhibits strong fluorescence upon binding to nucleic acids and has sufficient precision to measure differences down to single cells. As shown in Figure 10, the presence of compound 1, a PDE1 inhibitor, showed a significant inhibitory effect on the transmembrane motility of BV2 cells, providing further evidence that compound 1 suppresses the release of pro-inflammatory markers.

[0136] Example 3: Detection of inflammatory biomarkers using a mouse zymon pleuritis model Zymosan was injected into the pleural cavity of mice to induce sterile inflammation using the method described in Example 1. Compound 1 was administered to the test subjects, and its effects on various inflammatory biomarkers were observed. Results were recorded after 4 hours. The subjects showed a clear decrease in cytokine markers after administration of compound 1. IFNγ, IL-1β, MCP1-β, and TNFα decreased in all serum and plasma samples after administration of compound 1. IL10 showed a decrease in serum.

[0137] Lipids are known to be involved in regulating numerous cellular responses, including cell growth and death, as well as inflammation / infection, via receptor-mediated pathways. Various lipids are involved in both the initiation and resolution of inflammation. Pro-resolving lipid mediators are naturally produced in the body from unsaturated fatty acids such as arachidonic acid (AA) and docosahexaenoic acid (DHA). Further research was conducted to identify metabolites of AA and DHA, which are summarized in Tables 2 and 3 below.

[0138] [Table 2]

[0139] [Table 3]

[0140] Regarding AA metabolism, as shown above, 12-HETE and 15-HETE, both intermediate mediators that contribute to the resolution of inflammation, were increased in frequency compared to the control, while TXB2, PGE2, and LTB4, all pro-inflammatory mediators, were decreased. In DHA metabolism, 17-HDOHE, RVD5, and 14-HDOHE, all involved in inflammation resolution, were increased. This lipid biomarker profile suggests that the test compound induces metabolites in the 15-LOX and 12-LOX pathways, indicating the recruitment of pathways that promote resolution. Furthermore, the test compound was shown not to induce metabolites in the pro-inflammatory pathway of 5-LOX. This application also includes the following aspects. [Aspect 1] A method for promoting the resolution of inflammation for the treatment or prevention of inflammation or inflammation-related diseases, comprising administering a PDE1 inhibitor to a patient in need. [Aspect 2] The patient may have bacterial infections (e.g., infections with Salmonella typhi, Salmonella typhimurium, Listeria monocytogenes, Mycobacterium tuberculosis, Mycobacterium ulcerans, and Mycobacterium avium); viral infections (e.g., African Swine Fever Virus, Classical Swine Fever Virus, Dengue Virus, Foot and Mouth Disease Virus, Human Immunodeficiency Virus (HIV) (e.g., HIV1), Influenza A virus) Viruses, Porcine Circovirus-2, Porcine Reproductive and Respiratory Syndrome Virus, Porcine Pseudorabies Virus, Respiratory Syncytial Virus, Severe Acute Respiratory Syndrome Coronavirus, West Nile Virus, Viral Hepatitis (e.g., Hepatitis A, Hepatitis B, Hepatitis C); Parasitic Infections (e.g., Taenia crassiceps, Toxoplasma gondii, Leishmania infantum, Schistosoma mansoni) Mansoni's infections; atopic dermatitis; pneumonia; cardiovascular diseases, e.g., atherosclerosis; obesity and insulin resistance; asthma; pulmonary fibrosis;The method according to embodiment 1, relating to a macrophage-mediated disease or disorder selected from cardiac obstructive pulmonary disease (COPD); neuropathic pain; stroke; diabetes mellitus; sepsis; non-alcoholic steatohepatitis (NASH); autoimmune hepatitis; systemic lupus erythematosus (SLE); wound healing; pleurisy; peritonitis; and cystic fibrosis. [Aspect 3] The patient, a. Elevated levels of one or more pro-inflammatory cytokines (e.g., selected from IL1β, TNFα, Ccl2, IL-6, and combinations thereof); b. A decrease in the level of one or more anti-inflammatory cytokines (e.g., IL-10); c. Increased level of M1 phenotype macrophages compared to M2 phenotype macrophages. The method according to any of the above embodiments, having the following characteristics. [Aspect 4] PDE1 inhibitors (A) Formula I, encompassing enantiomers, diastereomers and racemates, in free form, salt form or prodrug form: [ka] [In the formula, (i) R 1 is H or C 1-4 It is alkyl (for example, methyl); (ii) R 4 is H or C 1-4 It is alkyl, R 2 and R 3 H or C 1-4 Alkyl (For example, R 2 and R 3 Both are methyl, or R 2 H is R 3 It is isopropyl, aryl, heteroaryl, (possibly hetero)arylalkoxy, or (possibly hetero)arylalkyl; or R 2 H is R 3 and R 4 They combine to form dimethylene bridges, trimethylene bridges, or tetramethylene bridges. (preferably, R 3 and R 4 They have a cis configuration together, for example, R 3 and R 4 The carbon atoms supporting the molecule each have an R configuration and an S configuration. (iii) R 5 is, for example, a substituted heteroarylalkyl, which is substituted with a haloalkyl; or R 5 It combines with one of the nitrogen atoms in the pyrazoro portion of formula I, and formula A:

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Claims

1. A pharmaceutical agent for the treatment of inflammation in patients requiring treatment of inflammation, comprising an amount of a PDE1 inhibitor effective in promoting macrophage activation from an M1 activated state to an M2 activated state in patients requiring treatment of inflammation, The patient may have a bacterial infection selected from infections caused by Salmonella typhi, Salmonella typhimurium, Listeria monocytogenes, Mycobacterium tuberculosis, Mycobacterium ulcerans, and Mycobacterium avium; African Swine Fever Virus, Classical Swine Fever Virus, Dengue Virus, Foot and Mouth Disease Virus, Human Immunodeficiency Virus (HIV) (e.g., HIV-1), Influenza A Virus, Porcine Circovirus (Porcelain) Viral infections selected from Circovirus-2, Porcine Reproductive and Respiratory Syndrome Virus, Porcine Pseudorabies Virus, Respiratory Syncytial Virus, Severe Acute Respiratory Syndrome Coronavirus, West Nile Virus, and viral hepatitis (e.g., Hepatitis A, Hepatitis B, Hepatitis C); parasitic infections selected from Taenia crassiceps, Toxoplasma gondii, Leishmania infantum, and Schistosoma mansoni; atopic dermatitis; pneumonia; obesity and insulin resistance; asthma; pulmonary fibrosis;A patient suffering from a macrophage-mediated disease or disorder selected from chronic obstructive pulmonary disease (COPD); diabetes mellitus; sepsis; non-alcoholic steatohepatitis (NASH); autoimmune hepatitis; systemic lupus erythematosus (SLE); pleurisy; peritonitis; and cystic fibrosis, the patient having elevated levels of M1 phenotype macrophages compared to M2 phenotype macrophages; The PDE1 inhibitor, (A) In free form or salt form, formula Ia: 【Chemistry 1】 [In the formula, (i) R 2 and R 3 Each of them is methyl, and R 4 and R 5 Each of these is H; (ii) R 6 is a phenylamino acid that may be halosubstituted; (iii) R 10 is a pyridyl that may be halo-substituted; (iv) X and Y are each C]; (B) Formula II in free form, salt form or prodrug form: 【Chemistry 2】 [In the formula, (i) X is methylene; (ii) Y is allirene; (iii) Z is aryl, pyridyl, or -C(O)-R 1 And; (iv) R 1 is C 1-6 alkyl, halo C 1-6 alkyl, -OH or -OC 1-6 alkyl (e.g., -OCH 3 ) and; (v) R 4 H is R 5 is an aryl which may be substituted with a halo; (vi) Here, Z may be replaced by a halo; (C) Formula III in free or salt form: 【Transformation 3】 [In the formula, (i) R 1 It is methyl; (ii) R 2 and R 3 Both are methyl; (iii) R 4 H is; (iv) R 5 is -C(=O)-C 1-6 Alkyl or C 1-6 An aryl which may be substituted with a hydroxyalkyl group; (v) R 6 H is R 7 is an aryl which may be substituted with a halo; (vi) n is 1] Selected from, Pharmaceuticals.

2. The patient, further, a. Elevated levels of one or more pro-inflammatory cytokines (e.g., selected from IL-1β, TNFα, Ccl2, IL-6, and combinations thereof); and b. A decrease in the level of one or more anti-inflammatory cytokines (e.g., IL-10). A pharmaceutical product according to claim 1, having the following characteristics.

3. PDE1 inhibitors, in free form or pharmaceutically acceptable form, include the following: 【Chemistry 4】 The pharmaceutical product according to claim 1 or 2.

4. PDE1 inhibitors, in free form or pharmaceutically acceptable form, include the following: 【Transformation 5】 The pharmaceutical product according to claim 1 or 2.

5. PDE1 inhibitors, in free form or pharmaceutically acceptable form, include the following: 【Transformation 6】 The pharmaceutical product according to claim 1 or 2.

6. The pharmaceutical product according to any one of claims 1 to 5, wherein a PDE1 inhibitor is administered in combination with a PDE4 inhibitor (for example, lolipram).

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