PDE1 inhibitors

Abstract

The present invention relates to a compound of formula (I) for use as a human PDE1 inhibitor, [Formula 1] JPEG2026534076000027.jpg39128In formula, R 1 , R 2 , R 3 , R 4 , and R 5 The invention provides compounds, or pharmaceutically acceptable salts thereof, as defined herein.

Description

[Technical Field]

[0001] The present invention relates to a specific human PDE1 inhibitor, a pharmaceutical composition comprising this compound, a method for treating a physiological disorder using this compound, and intermediates and processes useful for the synthesis of this compound. [Background technology]

[0002] Phosphodiesterases (PDEs) are enzymes that regulate the cellular levels of cyclic nucleotides cAMP and cGMP by controlling the rate at which these nucleotides are hydrolyzed. PDE1, a calcium and calmodulin-dependent PDE, is one of at least 11 known PDE families. PDE1 is expressed in many tissues, including the brain, heart, lungs, kidneys, and smooth muscle. In addition, PDE1 consists of a family of three known isoforms: PDE1A, PDE1B, and PDE1C.

[0003] Patients with diabetes often develop a form of chronic kidney disease called diabetic nephropathy (or diabetic nephropathy). It is estimated that diabetic nephropathy may affect 40% of people with diabetes. Treatment options for diabetic nephropathy are limited and include the use of blood pressure-lowering medications, blood glucose, diet, and weight management, as well as regular physical activity. Therefore, there is a need for additional treatment options for patients with chronic kidney disease, particularly diabetic nephropathy.

[0004] U.S. Patent No. 8,299,080 discloses certain quinoxaline derivatives for the treatment of various disorders, including urinary disorders and hypertension. In addition, European Patent No. 0040401 discloses certain substituted triazoloquinoxaline-4-ones having antihypertensive activity. Certain other PDE1 inhibitor compounds are disclosed in International Publications 2018 / 039051, 2017 / 139186, 2019 / 032383, 2019 / 156861, and 2019 / 027783.

[0005] There is still a need for PDE1 inhibitor therapy using compounds that exhibit desirable efficacy, minimize induction of cytochrome P450 (cyp) enzymes in patients, and minimize toxicity in patients, for example, with respect to undesirable cardiac effects. This invention provides certain novel compounds that are inhibitors of human PDE1. In addition, this invention provides certain novel compounds that are selective inhibitors of human PDE1A, PDE1B, and PDE1C compared to other human PDEs such as PDE3A, PDE4D, and PDE6AB. Furthermore, this invention provides certain novel compounds that may have antihypertensive effects and may also improve renal blood flow. In addition, certain compounds of this invention may alleviate renal fibrosis. [Overview of the Initiative] [Means for solving the problem]

[0006] Therefore, the present invention relates to a compound of formula I, [ka] In the formula, R 1 , R 2 , and R 3 However, each is independently a C1-C3 alkyl group that is optionally substituted with H, a halogen, or one or more halogens. R 4 However, it is H, a halogen, a C1-C4 alkyl group optionally substituted with one or more halogens, a cyclopropyl group, or a CH=CH-CH3 group. R 5 However, the compound is a C1-C3 alkyl or cyclopropyl compound that is optionally substituted with one or more halogens. Or provide a pharmaceutically acceptable salt thereof.

[0007] The PDE1 pathway is associated with conditions and diseases such as renal disease, cardiovascular disease, hypertension, pulmonary hypertension, Parkinson's disease, atherosclerosis, neurocognitive disorders, schizophrenia, cognitive and motor impairments associated with Alzheimer's disease, and attention deficit hyperactivity disorder (Samidurai A, et al., Pharmacol Ther. October 2021, 226:107858. doi:10.1016 / j.pharmthera.2021.107858).

[0008] Accordingly, the present invention also provides a method for treating diabetic nephropathy, chronic kidney disease, acute kidney injury, hypertension, resistant hypertension, Parkinson's disease, atherosclerotic cardiovascular disease, angina pectoris, myocardial infarction, heart failure, pulmonary hypertension, ischemic stroke, cognitive impairment associated with schizophrenia, cognitive impairment associated with Alzheimer's disease, motor impairment, or attention deficit and hyperactivity disorder in a patient, comprising administering an effective amount of a compound of formula I, or a pharmaceutically acceptable salt thereof, to a patient in need of such treatment.

[0009] In addition, the present invention provides compounds of formula I or pharmaceutically acceptable salts thereof for use in therapy.

[0010] The present invention further provides compounds of formula I or pharmaceutically acceptable salts thereof for use in the treatment of diabetic nephropathy, chronic nephropathy, acute kidney injury, hypertension, resistant hypertension, Parkinson's disease, atherosclerotic cardiovascular disease, angina pectoris, myocardial infarction, heart failure, pulmonary hypertension, ischemic stroke, cognitive impairment associated with schizophrenia, cognitive impairment associated with Alzheimer's disease, motor impairment, or attention deficit hyperactivity disorder.

[0011] Furthermore, the present invention provides the use of compounds of formula I or pharmaceutically acceptable salts thereof for the manufacture of pharmaceuticals for the treatment of diabetic nephropathy, chronic nephropathy, acute kidney injury, hypertension, resistant hypertension, Parkinson's disease, atherosclerotic cardiovascular disease, angina pectoris, myocardial infarction, heart failure, pulmonary hypertension, ischemic stroke, cognitive impairment associated with schizophrenia, cognitive impairment associated with Alzheimer's disease, motor impairment, or attention deficit hyperactivity disorder.

[0012] The present invention further provides a pharmaceutical composition comprising a compound of Formula I or a pharmaceutically acceptable salt thereof, together with one or more pharmaceutically acceptable carriers, diluents, or excipients. The present invention further provides a process for preparing a pharmaceutical composition, which comprises mixing a compound of Formula I or a pharmaceutically acceptable salt thereof with one or more pharmaceutically acceptable carriers, diluents, or excipients. The present invention also encompasses novel intermediates and processes for the synthesis of the compound of Formula I. [Mode for Carrying Out the Invention]

[0013] As used herein, the terms "treating", "therapy", or "treat" include suppressing, restraining, delaying, arresting, or reversing the progression or severity of an existing symptom or disorder.

[0014] As used herein, the term "patient" refers to a mammal such as a dog or a human, with a human being preferred.

[0015] As used herein, the term "effective amount" refers to an amount or dose of the compound of the present invention or a pharmaceutically acceptable salt thereof that, when administered to a patient in single or multiple doses, provides the desired effect to the patient under diagnosis or treatment.

[0016] As used herein, "C1-C3 alkyl" refers to methyl, ethyl, n-propyl, and isopropyl, and "C1-C4 alkyl" refers to methyl, ethyl, n-propyl, isopropyl, n-butyl, s-butyl, t-butyl, and isobutyl.

[0017] As used herein, "halogen" refers to F, Cl, Br, and I. Examples of halogen-substituted alkyl include trifluoromethyl, chloromethyl and the like.

[0018] The effective dose can be readily determined by those skilled in the art by using known techniques and by observing results obtained under similar circumstances. Numerous factors, considered by those skilled in the art, when determining the effective dose for a patient, include, but are not limited to, the patient's size, age, and overall health status; the specific disease or disorder involved; the degree or involvement or severity of the disease or disorder; the individual patient's response; the specific compound administered; the mode of administration; the bioavailability characteristics of the administered preparation; the dose regimen chosen; the use of concomitant medications; and other relevant circumstances.

[0019] The compounds of the present invention are formulated as pharmaceutical compositions to be administered by any route that makes the compounds bioavailable, including oral and parenteral routes. More preferably, such compositions are for oral administration. Such pharmaceutical compositions and processes for preparing them are well known in the art (e.g., Remington: The Science and Practice of Pharmacy, LV Allen, Editor, 22 nd (See Edition, Pharmaceutical Press, 2012.)

[0020] Compounds for use according to the methods disclosed herein may be administered as a single compound or in combination of compounds. Compounds and compositions disclosed herein may be administered by therapeutic methods known in the art. Accordingly, various such compounds and compositions may be administered in any preferred manner in combination with such methods. For example, administration may include oral, intravenous, intra-arterial, intramuscular, subcutaneous, intraperitoneal, parenteral, transdermal, vaginal, intranasal, mucosal, sublingual, topical, rectal, or subcutaneous administration, or any combination thereof.

[0021] Compounds of formula I are particularly useful in the therapeutic methods of the present invention, but certain groups, substituents, and compounds are preferred. These preferred groups, substituents, and compounds are described in the following paragraphs. It will be understood that these preferences are applicable to both the therapeutic methods and novel compounds of the present invention.

[0022] R 1 , R 2 , and R 3 are preferably H.

[0023] R 4 is preferably C1-C4 alkyl.

[0024] R 5 is preferably C1-C3 alkyl.

[0025] R 4 is more preferably ethyl.

[0026] R 5 is more preferably methyl.

[0027] It is further preferred that the compound of Formula I is in free base form.

[0028] R 1 , R 2 , and R 3 are H, R 4 is particularly preferably ethyl.

[0029] R 1 , R 2 , and R 3 are H, R 5 is particularly preferably methyl.

[0030] R 4 is ethyl, R 5 is particularly preferably methyl.

[0031] Examples of the compounds disclosed in the present specification include [Chemical Formula] [Chemical Formula] or a pharmaceutically acceptable salt thereof.

[0032] Compounds of the following formula [ka] Most preferably, the pharmaceutically acceptable salt thereof, and most preferably, the free base of the compound described immediately above.

[0033] A pharmaceutically acceptable salt of the compound of the present invention can be formed, for example, by the reaction of a suitable free base of the compound of the present invention with a suitable pharmaceutically acceptable acid in a suitable solvent under standard conditions well known in the art. For example, this can be found in Gould, PL, "Salt selection for basic drugs," International Journal of Pharmaceutics, 33:201-217 (1986), Bastin, RJ, et al., "Salt Selection and Optimization Procedures for Pharmaceutical New Chemical Entities," Organic Process Research and Development, 4:427-435 (2000), and Berge, SM, et al., "Pharmaceutical Salts," Journal of Pharmaceutical Sciences, 66:1-19 (1977).

[0034] Individual isomers, enantiomers, and diastereomers can be separated or degraded by those skilled in the art at any convenient point in the synthesis of the compounds of the present invention by methods such as selective crystallization techniques or chiral chromatography (see, for example, J. Jacques, et al., "Enantiomers, Racemates, and Resolutions," John Wiley and Sons, Inc., 1981, and ELEliel and SHWilen, "Stereochemistry of Organic Compounds," Wiley-Interscience, 1994). The names "Isomer 1" and "Isomer 2" refer to the compounds that elute from chiral chromatography under the first and second specific conditions, respectively.

[0035] Certain abbreviations are defined as follows: "ACN (acetonitrile)" refers to acetonitrile, "AcOH" refers to glacial acetic acid, "DBU" refers to 1,8-diazabicyclo[5.4.0]undec-7-ene, "DCM" refers to dichloromethane or methylene chloride, "DIPEA" refers to N,N-diisopropylethylamine, "DMF" refers to N,N-dimethylformamide, "DMSO" refers to dimethyl sulfoxide, "EDCI" refers to 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, and "ES / MS" refers to electrospray mass. In liquid chromatography-mass spectrometry, "siRNA" refers to ethyl acetate, "Et2O" refers to diethyl ether, "EtOH" refers to ethanol, "HMDS" refers to heamethyldisilazane, "HOBT" refers to hydroxybenzotriazole, "IPA" refers to isopropanol, "TFA" refers to trifluoroacetic acid, "HATU" refers to 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxidehexafluorophosphate, "LCMS" refers to liquid chromatography-mass spectrometry, "RBF" refers to a round-bottom flask, "TR" refers to retention time, "hr" refers to time, and "IC" refers to a time interval. 50" refers to the concentration of the drug that produces 50% of the maximum possible inhibitory response for that drug, "μmol" refers to micromoles or micromoles, "min" refers to minutes, "MeOH" refers to methanol or methyl alcohol, "MTBE" refers to methyl tert-butyl ether, "NiNTA" refers to chromatography using an agarose stationary phase functionalized with nitrilotriacetic acid as a chelating agent, "POCl3" refers to phosphorus oxychloride, "RT" refers to room temperature, "SNAr" refers to a nucleophilic aromatic substituent, "TEA" refers to triethylamine, "THF" refers to tetrahydrofuran, "Tris" refers to 2-amino-2-hydroxymethyl-propane-1,3-diol, "U / ml" refers to units per milliliter, "wt" refers to weight, and "Pd(OAc)2" refers to palladium(II) acetate.

[0036] The compounds of the present invention can be prepared by various procedures known to those skilled in the art, some of which are illustrated in the following schemes, preparations, and examples. Those skilled in the art will recognize that, in order to prepare the compounds of the present invention, specific synthesis steps for each of the described routes may be combined in different ways or in combination with steps of different schemes. The products of each of the following steps can be recovered by conventional methods well known in the art, including extraction, evaporation, precipitation, chromatography, filtration, grinding, and crystallization. In the following schemes, all substituents are as previously defined unless otherwise indicated. Reagents and starting materials are readily available to those skilled in the art. To further illustrate the present invention without limiting its scope, the following representative schemes, preparations, and examples are provided.

[0037] Scheme 1 [ka]

[0038] Preparation 1 Synthesis of 2-fluoro-1-nitro-3-vinylbenzene [ka]

[0039] Scheme 1, Process A 1-Bromo-2-fluoro-3-nitrobenzene (9.500 g, 43.182 mmol), 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (9.976 g, 64.773 mmol), S-phos (1.772 g, 4.318 mmol), and potassium phosphate, tribasic, N-hydrate (18.332 g, 86.364 mmol) were combined in a mixture of 1,4-dioxane (100 mL, 1.171 mol) and water (10 mL, 555.084 mmol). The mixture was purged with nitrogen for 30 minutes. Pd(OAc)2 (969.4805 mg, 4.318 mmol) was added, and the mixture was then heated to 80°C over the weekend. The mixture was cooled to ambient temperature and poured through filter paper. The filtrate was then partitioned between ethyl acetate and brine. The aqueous layer was removed, the organic layer was washed once with brine, dried over sodium sulfate, and then reduced to brown oil. The material was purified on silica gel using 5% ethyl acetate / hexane as the solvent to obtain orange oil as the title compound (7.2 g, quantitative yield).

[0040] Preparation 2 Synthesis of N-(4-methoxybenzyl)-2-nitro-6-vinylaniline [ka]

[0041] Scheme 1, Process B2-Fluoro-1-nitro-3-vinylbenzene (7.22 g, 43.197 mmol) and 4-methoxybenzenemethaneamine (11.851 g, 86.395 mmol) were combined in ethanol (150 mL, 2.576 mol), and the mixture was heated under reflux overnight. LC-MS indicated that the reaction was complete. The mixture was cooled to ambient temperature, and the solvent was removed under vacuum. The residue was partitioned between ethyl acetate and water. The aqueous layer was removed, and the organic layer was washed once with water and once with brine, dried over sodium sulfate, and then reduced to oil. The material was purified on silica gel using 5% ethyl acetate / hexane as the solvent to obtain orange oil as the title compound (11.73 g, 96% yield).

[0042] Preparation 3 6-ethyl-N 1 Synthesis of (4-methoxybenzyl)benzene-1,2-diamine [ka]

[0043] Scheme 1, Process C To a 2250 ml Parr shaker bottle, N-(4-methoxybenzyl)-2-nitro-6-vinylaniline (11.73 g, 41.257 mmol), 5% Pt / C (2.65 g, 11.670 mmol), and ethyl acetate (1000 mL) were added. The reaction vessel was placed on a shaker at 60 psig at room temperature for 4 hours. LC-MS showed product formation. The reaction mixture was filtered and rinsed with ethyl acetate. The solvent was removed under vacuum to obtain a yellow oil as the title compound (9.94 g, 89% yield). MS (m / z): 257.1 (M+1).

[0044] Preparation 4 Synthesis of 8-ethyl-1-(4-methoxybenzyl)-1,4-dihydroquinoxaline-2,3-dione [ka]

[0045] Scheme 1, Process D :6-ethyl-N 1 -(4-methoxybenzyl)benzene-1,2-diamine (9.94 g, 36.875 mmol) was dissolved in ethanol (270 mL, 4.638 mol). Then, sodium ethoxide (25.093 g, 77.438 mmol) and diethyl oxalate (10.778 g, 73.751 mmol) were added. The reaction mixture was stirred overnight at room temperature. LC-MS indicated that the reaction was complete. The solvent was then removed under vacuum to obtain a yellowish-brown foam. This foam was sonicated in water and then stirred at ambient temperature for 2 hours. The resulting solid was isolated by vacuum filtration, washed with water, dried for 30 minutes, and then placed overnight in a vacuum oven at 50°C to obtain a yellow solid as the title compound (10.91 g, 95% yield). MS (m / z): 309.0 (M-1).

[0046] Preparation 5 Synthesis of 3-chloro-8-ethyl-1-(4-methoxybenzyl)quinoxaline-2(1H)-one [ka]

[0047] Scheme 1, Process E 8-ethyl-1-(4-methoxybenzyl)-1,4-dihydroquinoxaline-2,3-dione (6.00 g, 19.333 mmol) was combined with thionyl chloride (12 mL, 164.713 mmol) and dimethylformamide (3 mL, 38.798 mmol) in toluene (150 mL, 1.418 mol). The mixture was heated at 115°C for 45 minutes. LC-MS showed that the starting material had been consumed. Toluene was then removed under vacuum. The resulting residue was dissolved in toluene and removed again under vacuum to obtain brown oil as the title compound (6.36 g, quantitative yield). The material was proceeded to the next stage without further purification. MS (m / z): 329.0 (M+1).

[0048] Preparation 6 Synthesis of 8-ethyl-3-hydrazineyl-1-(4-methoxybenzyl)quinoxaline-2(1H)-one [ka]

[0049] Scheme 1, Process F 3-Chloro-8-ethyl-1-(4-methoxybenzyl)quinoxaline-2(1H)-one (6.355 g, 19.33 mmol) was suspended in ethanol (80 mL, 1.374 mol), and hydrazine (3.097 mL, 96.651 mmol) was added. The mixture was heated under reflux for 3 hours and then cooled to ambient temperature. The reaction mixture was stirred over the weekend. The resulting precipitate was isolated by vacuum filtration. The cake was washed with ethanol and dried for 2 hours to obtain a yellow solid as the title compound (4.23 g, 67% yield). MS (m / z): 325.2 (M+1).

[0050] Preparation 7 Synthesis of N'-(5-ethyl-4-(4-methoxybenzyl)-3-oxo-3,4-dihydroquinoxaline-2-yl)-1-methylcyclopropane-1-carbohydrazide [ka]

[0051] Scheme 1, Process G8-Ethyl-3-hydrazineyl-1-(4-methoxybenzyl)quinoxaline-2(1H)-one (4.23 g, 13.04 mmol), 1-methylcyclopropane-1-carboxylic acid (22.219 g, 22.168 mmol), HATU (8.429 g, 22.168 mmol), and diisopropylethylamine (7.960 mL, 45.640 mmol) were combined in dimethylformamide (80 mL, 1.035 mol) and stirred overnight at ambient temperature. The mixture was partitioned between ethyl acetate and water. The aqueous layer was removed, and the organic layer was washed twice with saturated sodium bicarbonate, then twice with brine, dried over sodium sulfate, and then reduced to the residue. The residue was purified on silica gel using 50% ethyl acetate / hexane as the solvent to obtain the title compound as yellow foam (3.0 g, 56% yield). MS(m / z): 407.2 (M+1).

[0052] Example 1 Synthesis of 6-ethyl-1-(1-methylcyclopropyl)-[1,2,4]triazolo[4,3-a]quinoxaline-4(5H)-one [ka]

[0053] Scheme 1, Process H:N'-(5-ethyl-4-(4-methoxybenzyl)-3-oxo-3,4-dihydroquinoxaline-2-yl)-1-methylcyclopropane-1-carbohydrazide (3.00 g, 7.380 mmol) was dissolved in acetic acid (30 mL, 523.541 mmol) and heated overnight at 115°C. LC-MS showed that the starting material had been consumed. The reaction mixture was cooled to ambient temperature and stirred overnight. The acetic acid was removed under vacuum. The resulting yellow semi-solid was dissolved in TFA (8 ml) and heated in a microwave at 90°C for 1 hour. The mixture was allowed to stand overnight at ambient temperature. The TFA was removed under vacuum, and the residue was sonicated in diethyl ether. The resulting solid was isolated by vacuum filtration. The cake was washed with ether and dried for 4 hours to obtain an off-white solid. The material was purified using silica gel with 70% ethyl acetate / hexane to 100% ethyl acetate to obtain an off-white solid as the title compound (1.22 g, 62% yield). MS (m / z): 269.0 (M+H), Molecular weight: 268.

[0054] PDE protein synthesis Nucleotide sequences encoding full-length human PDE1A (NP_001003683.1), PDE1C (NP_005011.1), PDE5A (NP_001074.2), PDE7B (NP_061818.1), and PDE9A (NP_002597.1) are inserted into a pFastBac1 (Invitrogen) vector with an N-terminal HIS tag. Nucleotide sequences encoding the catalytic domains (residues 641-1141) of full-length human PDE4D (NP_006194.2) and PDE3A (NP_000912.3) are inserted into a pFastBac1 (Invitrogen) vector with a C-terminal HIS tag. Nucleotide sequences encoding full-length human PDE8A (NP_002596.1) and PDE11A (AAI12394.1) are inserted into a pFastBac1 (Invitrogen) vector with an N-terminal Flag tag. Nucleotide sequences encoding full-length human PDE10A (AAD32595.1) are inserted into a pFastBac1 (Invitrogen) vector with a C-terminal Flag-His tag. Nucleotide sequences encoding full-length human PDE6A (NP_000431.2) and PDE6B (AAH00249.1) are inserted into pFastBacDual (Invitrogen) vectors with an N-terminal HIS tag and an N-terminal Flag tag, respectively, to produce PDE6A / 6B dimers. Baculovirus production and protein expression in Sf9 cells are performed using the Bac-to-Bac baculovirus expression system (Invitrogen) protocol. Nucleotide sequences encoding full-length human PDE1B (NP_000915.1) and PDE2A (NP_002590.1) were inserted into pIEX4 (Novagen) with a C-terminal HIS tag, and both proteins were produced in Sf9 cells according to the vendor's protocol (Novagen). The His-tagged PDE proteins were purified using size exclusion chromatography on a SUPERDEX® 200 column (GE Healthcare) in storage buffer (20 mM Tris HCl, pH 7.5, 150 mM NaCl, 10% glycerol) after Ni-NTA agarose (Qiagen).Flag-tagged PDE proteins, including PDE6A / 6B, are purified using anti-Flag M2-agarose (Sigma), followed by NiNTA column chromatography, and then eluted in storage buffer (50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 10% glycerol, 0.1 mg / ml Flag peptide). All purified proteins are stored in small, fixed volumes at -80°C.

[0055] Phosphodiesterase Enzyme assay All 3',5' cyclic nucleotide phosphodiesterase (PDE) enzyme activity is measured using a radioenzyme assay (scintillation proximity assay) based on an SPA detection system. The test compound is diluted in pure dimethyl sulfoxide (DMSO) using a 10-point concentration response curve. The maximum compound concentration in the reaction mixture is 10 or 100 μM. After pre-incubating the appropriate concentration of the compound with one of the PDE enzymes for 30 minutes, the reaction is initiated by adding the substrate. The reaction is allowed to proceed at room temperature for 60 minutes. The reaction is then stopped by adding SPA beads. The sample is read after 12 hours using a MICROBETA® TRILUX® counter. 50 IC refers to the concentration of a compound that produces 50% of the maximum possible inhibitory response for that compound. 50 The value is calculated by plotting normalized data against log[compound] and fitting the data using the four-parameter logistic equation.

[0056] Ca 2+ - Calmodulin-dependent PDE enzyme assay PDE1B, PDE1A, and PDE1C are cloned and purified according to a standard protein synthesis procedure. Assay buffer is prepared at pH 7.5 to obtain the final concentrations in an assay using 50 mM Tris HCl, 50 mM MgCl2, 4 mM CaCl2, 0.1% bovine serum albumin, and 6 U / ml calmodulin in water. The final enzyme concentrations are 0.25, 0.074, and 0.0012 nM for PDE1A, PDE1B, and PDE1C, respectively. The reaction is carried out with the substrate [3 The process is initiated by adding [H]cAMP, and a final concentration of 47 nM is obtained.

[0057] [Table 1]

[0058] The data in Table 1 demonstrate that the compound from Example 1 inhibits human PDE1A, PDE1B, and PDE1C enzyme activity in vitro.

[0059] [ 3 PDE enzyme assay using [H]cAMP as a substrate The following phosphodiesterase activities, human PDE3A (catalytic domain), human PDE4D, human PDE7B, and human PDE8A, [ 3 The enzymes are measured using [H]cAMP as the reaction substrate. All of these enzymes are cloned and purified according to standard procedures. Assay buffers are prepared at pH 7.5 to obtain the final concentrations in the assay of 50 mM Tris-HCl, 8.3 mM MgCl2, 1.7 mM ethylenediaminetetraacetic acid (EDTA), and 0.1% bovine serum albumin. The final enzyme concentrations are 0.008, 0.021, 0.5, and 0.06 nM for PDE3A, PDE4D, PDE7B, and PDE8A, respectively. The reaction is carried out with the substrate [ 3 The process is initiated by adding [H]cAMP, and a final concentration of 47 nM is obtained.

[0060] [Table 2]

[0061] [ 3 PDE enzyme assay using [H]cGMP as a substrate The following phosphodiesterase activities, human PDE2A, human PDE5A, human PDE6A / 6B, human PDE9A, human PDE10A, and human PDE11A, [ 3The reaction substrate is cGMP (H) used for measurement. The catalytically active form of human PDE6 is a dimer composed of α (human PDE6A) and β subunit (human PDE6B). The human PDE6A / 6B dimer is produced by expression and purification strategies using two purification steps: NiNTA and anti-FLAG Sepharose chromatography. The remaining enzyme is cloned and purified in-house according to standard procedures. Assay buffers are prepared to obtain the final concentrations in the assay using 50 mM Tris HCl, 8.3 mM MgCl2, 1.7 mM EDTA, and 0.1% bovine serum albumin at pH 7.5. The final enzyme concentrations are 0.2, 0.002, 5, 1, 0.03, and 0.03 nM for human PDE2A, human PDE5A, human PDE6AB, human PDE9A, human PDE10A, and human PDE11A, respectively. The reaction involves the substrate [ 3 The assay is initiated by adding [H]cGMP, and for human PDE2A, human PDE10A, human PDE5A, human PDE6AB, and human PDE11A assays, a final concentration of 80 nM is obtained, but for human PDE9A, 20 nM is used. 3 Use H]cGMP.

[0062] [Table 3]

[0063] The data in Tables 1, 2, and 3 demonstrate that the compound of Example 1 is a selective inhibitor of human PDE1A, PDE1B, and PDE1C in vitro, compared to human PDE2A, PDE3A, PDE4D, PDE5A, PDE6AB, PDE7B, PDE8A, PDE9A, PDE10A, and PDE11A.

[0064] Example 2 Other compounds were prepared in the same manner as in Example 1, as shown below.

[0065] [Table 4]

[0066] Table 5

[0067] Table 6

[0068] Table 7

[0069] Table 8

Claims

1. The following compound is given by the formula: 【Chemistry 1】 In the formula, R １ , R ２ , and R ３ However, each is independently a C1-C3 alkyl group that is optionally substituted with H, a halogen, or one or more halogens. R ４ However, H, halogens, C1-C4 alkyl groups optionally substituted with one or more halogens, cyclopropyl groups, or CH=CH-CH ３ And, R ５ However, the compound is a C1-C3 alkyl or cyclopropyl compound that is optionally substituted with one or more halogens. or a pharmaceutically acceptable salt thereof.

2. R １ , R ２ , R ３ is H, the compound or salt according to claim 1.

3. R ４ The compound or salt according to claim 1 or 2, wherein the compound is a C1-C4 alkyl group.

4. R ４ The compound or salt according to any one of claims 1 to 3, wherein the compound is ethyl.

5. R ５ The compound or salt according to any one of claims 1 to 4, wherein the compound is a C1 to C3 alkyl group.

6. R ５ The compound or salt according to any one of claims 1 to 5, wherein the compound is methyl.

7. The compound or salt according to any one of claims 1 to 6, wherein the compound is in the form of a free base.

8. The aforementioned compound, 【Chemistry 2】 The compound or salt according to claim 1, or a pharmaceutically acceptable salt thereof.

9. The aforementioned compound, 【Transformation 3】 The compound or salt according to claim 8. 【Request Item 10】 【Chemistry 4】 The compound or salt according to claim 8, which is a pharmaceutically acceptable salt of the compound.

11. A method for treating diabetic nephropathy, chronic nephropathy, acute kidney injury, hypertension, resistant hypertension, Parkinson's disease, atherosclerotic cardiovascular disease, angina pectoris, myocardial infarction, heart failure, pulmonary hypertension, ischemic stroke, cognitive impairment associated with schizophrenia, cognitive impairment associated with Alzheimer's disease, motor impairment, or attention deficit and hyperactivity disorder in a patient, comprising administering an effective amount of a compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof, to a patient in need thereof.

12. A compound according to any one of claims 1 to 10 or a pharmaceutically acceptable salt thereof, for use in therapy.

13. A compound according to any one of claims 1 to 10 or a pharmaceutically acceptable salt thereof for use in the treatment of diabetic nephropathy, chronic nephropathy, acute kidney injury, hypertension, resistant hypertension, Parkinson's disease, atherosclerotic cardiovascular disease, angina pectoris, myocardial infarction, heart failure, pulmonary hypertension, ischemic stroke, cognitive impairment associated with schizophrenia, cognitive impairment associated with Alzheimer's disease, motor impairment, or attention deficit hyperactivity disorder.

14. Use of a compound according to any one of claims 1 to 10 or a pharmaceutically acceptable salt thereof for the manufacture of a pharmaceutical product for the treatment of diabetic nephropathy, chronic nephropathy, acute kidney injury, hypertension, resistant hypertension, Parkinson's disease, atherosclerotic cardiovascular disease, angina pectoris, myocardial infarction, heart failure, pulmonary hypertension, ischemic stroke, cognitive impairment associated with schizophrenia, cognitive impairment associated with Alzheimer's disease, motor impairment, or attention deficit hyperactivity disorder.

15. A pharmaceutical composition comprising a compound according to any one of claims 1 to 10 or a pharmaceutically acceptable salt thereof, together with one or more pharmaceutically acceptable carriers, diluents, or excipients.

16. A process for preparing a pharmaceutical composition, comprising mixing a compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof, with one or more pharmaceutically acceptable carriers, diluents, or excipients.

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