PDE1 inhibitors

Abstract

The present invention provides compounds of Formula I: wherein R1, R2, R3, R4 and R5 are each as defined herein, or pharmaceutically acceptable salts thereof, useful as human PDE1 inhibitors.

Description

[0001] This invention relates to certain human PDE1 inhibitors, pharmaceutical compositions comprising compounds, methods of using said compounds to treat physiological disorders, and intermediates and methods that can be used to synthesize said compounds.

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

[0003] Patients with diabetes often develop some form of chronic kidney disease, known as diabetic nephropathy (or diabetic kidney disease). It is estimated that diabetic nephropathy may affect up to 40% of people with diabetes. Treatment options for diabetic nephropathy are limited and include the use of medications to lower blood pressure, managing blood sugar levels, diet and weight, and regular physical activity. Therefore, patients with chronic kidney disease, especially diabetic nephropathy, require additional treatment options.

[0004] US Patent No. 8,299,080 discloses certain quinoxaline derivatives for the treatment of various disorders such as dysuria and hypertension. Furthermore, European Patent No. 0 404 401 discloses certain substituted triazoloquinoxaline-4-ones with antihypertensive activity. Several other PDE1 inhibitor compounds are disclosed in WO 2018 / 039051, WO 2017 / 139186, WO 2019 / 032383, WO 2019 / 156861, and WO 2019 / 027783.

[0005] There remains a need for compounds used in PDE1 inhibitor therapy that exhibit desirable potency, minimize induction of cytochrome P450 (cyp) enzymes in patients, and minimize toxicity to patients, such as regarding undesirable cardiac effects. This invention provides certain novel compounds as inhibitors of human PDE1. Furthermore, this invention provides certain novel compounds as selective inhibitors of human PDE1A, PDE1B, and PDE1C relative to other human PDEs (such as PDE3A, PDE4D, and PDE6AB). In addition, this invention provides certain novel compounds that can have antihypertensive effects and can also improve renal blood flow. Furthermore, certain compounds of this invention can reduce renal fibrosis.

[0006] Therefore, the present invention provides compounds of formula I: Where R 1 R 2 and R 3 Each is independently H, a halogen, or a C1-C3 alkyl group optionally substituted with one or more halogens; R 4 H, halogen, C1-C4 alkyl group optionally substituted with one or more halogens, cyclopropyl, or CH=CH-CH3; and R 5 It is a C1-C3 alkyl group optionally substituted with one or more halogens or cyclopropyl groups; Or its pharmaceutically acceptable salt.

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

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

[0009] Furthermore, 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 the treatment of diabetic nephropathy, chronic kidney disease, acute kidney injury, hypertension, refractory hypertension, Parkinson's disease, atherosclerotic cardiovascular disease, angina pectoris, myocardial infarction, heart failure, pulmonary hypertension, ischemic stroke, cognitive deficits associated with schizophrenia, cognitive deficits associated with Alzheimer's disease, motor disorders, or attention deficit and hyperactivity disorder.

[0011] Furthermore, the present invention provides the use of compounds of Formula I or pharmaceutically acceptable salts thereof in the manufacture of medicaments for treating diabetic nephropathy, chronic kidney disease, acute kidney injury, hypertension, refractory hypertension, Parkinson's disease, atherosclerotic cardiovascular disease, angina pectoris, myocardial infarction, heart failure, pulmonary hypertension, ischemic stroke, cognitive deficits associated with schizophrenia, cognitive deficits associated with Alzheimer's disease, movement disorders, or attention deficit and hyperactivity disorder.

[0012] The present invention further provides pharmaceutical compositions comprising a compound of formula I or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers, diluents, or excipients. The present invention further provides a method for preparing a pharmaceutical composition comprising 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 covers novel intermediates and methods for synthesizing compounds of formula I.

[0013] As used herein, the terms “treating,” “treatment,” or “to treat” include suppressing, curbing, slowing, stopping, or reversing the progression or severity of existing symptoms or disorders.

[0014] As used in this article, the term “patient” refers to a mammal, such as a dog or a human, with humans being preferred.

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

[0016] As used herein, “C1-C3 alkyl” refers to methyl, ethyl, n-propyl, and isopropyl; “C1-C4 alkyl” refers to methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, and isobutyl.

[0017] As used herein, "halogen" refers to F, Cl, Br, and I. Examples of alkyl groups substituted with halogens include trifluoromethyl, chloromethyl, etc.

[0018] Those skilled in the art can readily determine the effective dose by using known techniques and by observing results obtained under similar conditions. In determining the effective dose for a patient, those skilled in the art consider many factors, including but not limited to: the patient's body type, age, and general health condition; the specific disease or disorder involved; the extent or severity of the disease or disorder; the individual patient's response; the specific compound administered; the administration method; the bioavailability characteristics of the administered formulation; the chosen dosing regimen; the use of concomitant medication; and other relevant circumstances.

[0019] The compounds of the present invention are formulated into pharmaceutical compositions for administration via any route that makes the compound bioavailable, including oral and parenteral routes. Most preferably, such compositions are for oral administration. Such pharmaceutical compositions and methods of their preparation are well known in the art (see, for example, Remington: The Science and Practice of Pharmacy, edited by LV Allen, 22nd edition, Pharmaceutical Press, 2012).

[0020] The compounds used according to the methods disclosed herein can be administered as a single compound or in combination. The compounds and compositions disclosed herein can be administered using treatment methods known in the art. Therefore, various such compounds and compositions can be administered in any suitable manner in combination with such methods. For example, administration may include oral, intravenous, intra-arterial, intramuscular, subcutaneous, intraperitoneal, parenteral, transdermal, intravaginal, intranasal, mucosal, sublingual, topical, rectal, or subcutaneous administration, or any combination thereof.

[0021] Compounds of Formula I are particularly useful for the treatment methods of the present invention, but certain groups, substituents, and compounds are preferred. The following paragraphs describe such preferred groups, substituents, and compounds. It will be understood that these preferences apply both to the treatment methods of the present invention and to the novel compounds of the present invention.

[0022] Preferably, R 1 R 2 and R 3 For H.

[0023] Preferably, R 4 It is a C1-C4 alkyl group.

[0024] Preferably, R 5 It is a C1-C3 alkyl group.

[0025] A further preferred option is R 4 It is an ethyl group.

[0026] A further preferred option is R 5 It is a methyl group.

[0027] Further preferably, the compound of formula I is in the form of a free base.

[0028] Especially preferred is when R 1 R 2 and R 3 When it is H, R 4 It is an ethyl group.

[0029] Especially preferred is when R1 R 2 and R 3 When it is H, R 5 It is a methyl group.

[0030] Especially preferred is when R 4 When it is ethyl, R 5 It is a methyl group.

[0031] Examples of compounds disclosed herein include: and Or its pharmaceutically acceptable salt.

[0032] The following compounds: The most preferred compounds are pharmaceutically acceptable salts, with the free base of the compound directly present being the most preferred.

[0033] Pharmaceutically acceptable salts of the compounds of the present invention can be formed, for example, by reacting a suitable free base of the compounds of the present invention with a suitable pharmaceutically acceptable acid in a suitable solvent under standard conditions known in the art. See, for example, 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] Those skilled in the art can separate or resolve individual isomers, enantiomers, and diastereomers at any convenient point in the synthesis of the compounds of the present invention by means of methods such as selective crystallization or chiral chromatography (see, for example, J. Jacques et al., " Enantiomers, Racemates, and Resolutions John Wiley and Sons, Inc., 1981 and El Eliel and S.H. Wilen, “ Stereochemistry of Organic Compounds (Wiley-Interscience, 1994). The names "Isomer 1" and "Isomer 2" refer to the compounds eluted first and second by chiral chromatography under specified conditions, respectively.

[0035] Some abbreviations are defined as follows: “ACN” 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; “ES / MS” refers to electrospray mass spectrometry; “EtOAc” refers to ethyl acetate; “Et2O” refers to diethyl ether; “EtOH” refers to ethanol; “HMDS” refers to N-methyldisilazane; “HOBT” refers to hydroxybenzotriazole; “IPA” refers to isopropanol; “TFA” refers to trifluoroacetic acid; “HATU” refers to 1-[bis(dimethylamino)methylene]-1 H -1,2,3-triazolo[4,5- b Pyridinium 3-oxide hexafluorophosphate; "LCMS" refers to liquid chromatography-mass spectrometry; "RBF" refers to a round-bottom flask; "TR" refers to retention time; "hr" refers to one or more hours; "IC 50 "" refers to the concentration of the reagent that produces 50% of the maximum possible inhibitory response; "μmol" refers to one micromolar or a few micromolars; "min" refers to one minute or a few minutes; "MeOH" refers to methanol or methyl alcohol; "MTBE" refers to methyl tert-butyl ether; "NiNTA" refers to chromatography with an agarose stationary phase functionalized with nitrotriacetic acid as a chelating agent; "POCl3" refers to phosphorus oxychloride; "RT" refers to room temperature; "SNAr" refers to nucleophilic aromatic substitution; "TEA" refers to triethylamine; "THF" refers to tetrahydrofuran; "Tris" refers to 2-amino-2-hydroxymethyl-prop-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 the specific synthetic steps of the various routes can be combined in different ways, or combined with steps from different schemes, to prepare the compounds of the present invention. The products of the following steps can be recovered by conventional methods known in the art, including extraction, evaporation, precipitation, chromatography, filtration, preparation, and crystallization. In the following schemes, unless otherwise specified, all substituents are as defined above. Reagents and raw materials are readily available to those skilled in the art. The following representative schemes, preparations, and examples are provided to further illustrate the invention without limiting its scope.

[0037] Option 1 .

[0038] Preparation 1 Synthesis of 2-fluoro-1-nitro-3-vinylbenzene Scheme 1, Step A: Combine 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 tribasic N-hydrate (18.332 g, 86.364 mmol) in a mixture of 1,4-dioxane (100 mL, 1.171 mol) and water (10 mL, 555.084 mmol). Purge the mixture with nitrogen for 30 minutes. Pd(OAc)₂ (969.4805 mg, 4.318 mmol) was added and heated to 80 °C over the weekend. The mixture was cooled to ambient temperature and poured onto filter paper. The filtrate was then partitioned between ethyl acetate and brine. The aqueous solution was removed and the organic matter was washed once with brine (1X), dried over sodium sulfate, and then reduced to a brown oil. The material was purified on silica gel using 5% ethyl acetate / hexane as a solvent to give an orange oil as the title compound (7.2 g, quantitative yield).

[0039] Preparation 2 Synthesis of N-(4-methoxybenzyl)-2-nitro-6-vinylaniline Scheme 1, Step B: 2-fluoro-1-nitro-3-vinylbenzene (7.22 g, 43.197 mmol) and 4-methoxybenzene (11.851 g, 86.395 mmol) were combined in ethanol (150 mL, 2.576 mol) and the mixture was then heated under reflux overnight. LCMS 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 solution was removed and the organic matter was washed once with water, once with brine, dried over sodium sulfate, and then reduced to an oil. The material was purified on silica gel using 5% ethyl acetate / hexane as a solvent to give an orange oil as the title compound (11.73 g, 96% yield).

[0040] Preparation 3 6-Ethyl-N 1 Synthesis of 1,2-(4-methoxybenzyl)phenyl-1,2-diamine Scheme 1, Step C: 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 to a 2250 ml Parr shake flask. The reaction vessel was placed on a shaker at 60 psig for 4 hours at room temperature. LCMS showed that the product had formed. The reaction mixture was filtered and washed with ethyl acetate. The solvent was removed under vacuum to give a yellow oil as the title compound (9.94 g, 89% yield). MS (m / z): 257.1 (M+1).

[0041] Preparation 4 Synthesis of 8-ethyl-1-(4-methoxybenzyl)-1,4-dihydroquinoxaline-2,3-dione Option 1, Step D: 6-Ethyl-N 1-(4-methoxybenzyl)phenyl-1,2-diamine (9.94 g, 36.875 mmol) was dissolved in ethanol (270 mL, 4.638 mol). Sodium ethoxide (25.093 g, 77.438 mmol) and diethyl oxalate (10.778 g, 73.751 mmol) were then added. The reaction mixture was stirred overnight at ambient temperature. LC-MS showed that the reaction was complete. The solvent was then removed under vacuum to give a brown foam. The foam was sonicated in water and then stirred at ambient temperature for 2 hours. The resulting solid was separated by vacuum filtration, washed with water, dried for 30 minutes, and then placed in a vacuum oven at 50 °C overnight to give a yellow solid as the title compound (10.91 g, 95% yield) after drying. MS (m / z): 309.0 (M-1).

[0042] Preparation 5 Synthesis of 3-chloro-8-ethyl-1-(4-methoxybenzyl)quinoxalin-2(1H)-one Scheme 1, Step E: 8-Ethyl-1-(4-methoxybenzyl)-1,4-dihydroquinoxalin-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 min. LCMS showed that the starting material had been consumed. Toluene was then removed under vacuum. The resulting residue was taken up in toluene and removed again under vacuum to give a brown oil as the title compound (6.36 g, quantitative yield). This material was carried forward without further purification. MS (m / z): 329.0 (M+1).

[0043] Preparation of 6 Synthesis of 8-ethyl-3-hydrazino-1-(4-methoxybenzyl)quinoxaline-2(1H)-one Scheme 1, Step F: 3-Chloro-8-ethyl-1-(4-methoxybenzyl)quinoxalin-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, then cooled to ambient temperature. This reaction caused a sensation over the weekend. The resulting precipitate was separated by vacuum filtration. The filter cake was washed with ethanol and dried for 2 hours to give a yellow solid, the title compound (4.23 g, 67% yield). MS (m / z): 325.2 (m+1).

[0044] Preparation 7 Synthesis of N'-(5-ethyl-4-(4-methoxybenzyl)-3-oxo-3,4-dihydroquinoxalin-2-yl)-1-methylcyclopropane-1-carbonylhydrazine Scheme 1, Step G: 8-Ethyl-3-hydrazino-1-(4-methoxybenzyl)quinoxalin-2(1H)-one (4.23 g, 13.04 mmol), 1-methylcyclopropane-1-carboxylic acid (22.219 g, 22.168 mmol), HATU (8.429 g, 22.167 mmol), and diisopropylethylamine (7.960 mL, 45.640 mmol) were mixed in dimethylformamide (80 mL, 1.035 mol) and stirred overnight at ambient temperature. The mixture was partitioned between ethyl acetate and water. The aqueous solution was removed, the organic matter was washed twice with saturated sodium bicarbonate (2X), washed twice with brine, dried over sodium sulfate, and then reduced to a residue. The residue was purified on silica gel using 50% ethyl acetate / hexane as solvent to give the yellow, foamy title compound (3.0 g, 56% yield). MS(m / z): 407.2 (M+1).

[0045] Example 1 Synthesis of 6-ethyl-1-(1-methylcyclopropyl)-[1,2,4]triazolo[4,3-a]quinoxaline-4(5H)-one Scheme 1, Step H: N'-(5-ethyl-4-(4-methoxybenzyl)-3-oxo-3,4-dihydroquinoxolin-2-yl)-1-methylcyclopropane-1-carbonylhydrazine (3.00 g, 7.380 mmol) was dissolved in acetic acid (30 mL, 523.541 mmol) and heated to 115 °C overnight. LCMS showed that the starting material had been consumed. The reaction was cooled to ambient temperature and stirred overnight. Acetic acid was removed under vacuum. The resulting yellow semi-solid was dissolved in TFA (8 mL) and heated in a microwave oven at 90 °C for 1 hour. The mixture was allowed to stand overnight at ambient temperature. TFA was removed under vacuum, and the residue was sonicated in diethyl ether. The resulting solid was separated by vacuum filtration. The filter cake was washed with diethyl ether and dried for 4 hours to give an off-white solid. The substance was purified on silica gel using 70% ethyl acetate / hexane to 100% ethyl acetate to give an off-white solid, the title compound (1.22 g, 62% yield). MS (m / z): 269.0 (M+H), molecular weight: 268.

[0046] PDE protein formation 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) were inserted into the 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) were inserted into the pFastBac1 (Invitrogen) vector with a C-terminal HIS tag. Nucleotide sequences encoding full-length human PDE8A (NP_002596.1) and PDE11A (AAI12394.1) were inserted into the pFastBac1 (Invitrogen) vector with an N-terminal Flag tag. Nucleotide sequences encoding full-length human PDE10A (AAD32595.1) were inserted into the pFastBac1 (Invitrogen) vector with a C-terminal Flag-His tag. Nucleotide sequences encoding full-length human PDE6A (NP_000431.2) and PDE6B (AAH00249.1) were inserted into the pFastBacDual (Invitrogen) vectors with N-terminal HIS tags and N-terminal Flag tags, respectively, to generate PDE6A / 6B dimers. Baculovirus generation and protein expression in Sf9 cells were performed according to 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 the generation of both proteins in Sf9 cells was performed according to the supplier's protocol (Novagen). Ni-NTA agarose (Qiagen) was used, followed by SUPERDEX... ®His-labeled PDE proteins were purified by size exclusion chromatography on a 200 column (GE Healthcare) in storage buffer (20 mM Tris-HCl, pH 7.5, 150 mM NaCl, 10% glycerol). Flag-labeled PDE proteins, including PDE6A / 6B, were purified by NiNTA column chromatography and then eluted with anti-Flag M2-agarose (Sigma) in storage buffer (50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 10% glycerol, 0.1 mg / ml Flag peptide). All purified proteins were stored in aliquots at -80°C.

[0047] Phosphodiesterase assay All 3',5' cyclic nucleotide phosphodiesterase (PDE) activities were measured using a radioenzyme assay based on the SPA detection system (scintillation proximity assay). Test compounds were diluted in pure dimethyl sulfoxide (DMSO) using a ten-point concentration response curve. The maximum compound concentration in the reaction mixture was 10 or 100 µM. Compounds were pre-incubated with any PDE enzyme at an appropriate concentration for 30 min, then the reaction was initiated by adding the substrate. The reaction was allowed to proceed at room temperature for 60 min. The reaction was then stopped by adding SPA beads. The activity was measured at MICROBETA after 12 hours. TM TRILUX ® The sample is read from the counter. "IC" 50 "IC" refers to the concentration of the compound that produces a 50% maximum inhibition response. IC is calculated by plotting normalized data vs. log[compound] and fitting the data using a four-parameter logistic equation. 50 value.

[0048] Ca 2+ - Calmodulin-dependent PDE enzyme assay PDE1B, PDE1A, and PDE1C were cloned and purified according to a standard protein generation procedure. Detection buffers were prepared to obtain the final concentrations in water at pH 7.5 for detection, consisting of 50 mM Tris-HCl, 50 mM MgCl2, 4 mM CaCl2, 0.1% bovine serum albumin, and 6 U / ml calmodulin. The final enzyme concentrations for PDE1A, PDE1B, and PDE1C were 0.25, 0.074, and 0.0012 nM, respectively. The substrate was added... 3 The H]cAMP reaction was initiated to produce a final concentration of 47 nM.

[0049] Table 1: In vitro efficacy of Example 1 against human PDE1A, PDE1B and PDE1C.

[0050] The data in Table 1 confirm that the compound of Example 1 inhibits the activity of human PDE1A, PDE1B and PDE1C enzymes in vitro.

[0051] use[ 3 PDE enzyme detection with H]cAMP as a substrate use[ 3 [H]cAMP was used as a reaction substrate to measure the activities of the following phosphodiesterases: human PDE3A (catalytic domain), human PDE4D, human PDE7B, and human PDE8A. All these enzymes were cloned and purified according to standard procedures. Detection buffers were prepared to obtain the final concentrations in the assay at pH 7.5 of 50 mM Tris-HCl, 8.3 mM MgCl2, 1.7 mM EDTA, and 0.1% bovine serum albumin. The final enzyme concentrations for PDE3A, PDE4D, PDE7B, and PDE8A were 0.008, 0.021, 0.5, and 0.06 nM, respectively. The activity was measured by adding substrate [H]. 3 The H]cAMP reaction was initiated to produce a final concentration of 47 nM.

[0052] Table 2: In vitro efficacy of Example 1 on human PDE3A (catalytic domain), PDE4D, PDE7B and PDE8A.

[0053] use[ 3 PDE enzyme detection with H]cGMP as substrate use[ 3 H]cGMP was used as the reaction substrate to measure the activities of the following phosphodiesterases: human PDE2A, human PDE5A, human PDE6A / 6B, human PDE9A, human PDE10A, and human PDE11A. The catalytically active form of human PDE6 is a dimer composed of α (human PDE6A) and β subunits (human PDE6B). The human PDE6A / 6B dimer was generated using an expression and purification strategy (using two purification steps, namely NiNTA and anti-FLAG Sepharose chromatography). The remaining enzymes were cloned and purified internally according to standard procedures. Detection buffers were prepared to obtain the final concentrations in the assay at pH 7.5 of 50 mM Tris-HCl, 8.3 mM MgCl2, 1.7 mM EDTA, and 0.1% bovine serum albumin. The final enzyme concentrations of human PDE2A, human PDE5A, human PDE6AB, human PDE9A, human PDE10A, and human PDE11A were 0.2, 0.002, 5, 1, 0.03, and 0.03 nM, respectively. This was achieved by adding substrate […]. 3The reaction was initiated with H]cGMP to produce a final concentration of 80 nM as determined for human PDE2A, human PDE10A, human PDE5A, human PDE6AB, and human PDE11A, while for human PDE9A, 20 nM was used. 3 H]cGMP.

[0054] Table 3: In vitro potency of Example 1 against PDE2A, PDE5A, PDE6AB, PDE9A, PDE10A and PDE11A.

[0055] The data in Tables 1, 2 and 3 confirm that the compound of Example 1 is a selective inhibitor of human PDE1A, PDE1B and PDE1C in vitro relative to human PDE2A, PDE3A, PDE4D, PDE5A, PDE6AB, PDE7B, PDE8A, PDE9A, PDE10A and PDE11A.

[0056] Example 2 Other compounds were prepared in a manner similar to that of Example 1, as shown below. .

Claims

1. A compound of the following formula: Where R 1 R 2 and R 3 Each is independently H, a halogen, or a C1-C3 alkyl group optionally substituted with one or more halogens; R 4 H, halogen, C1-C4 alkyl group optionally substituted with one or more halogens, cyclopropyl, or CH=CH-CH3; and R 5 It is a C1-C3 alkyl group optionally substituted with one or more halogens or cyclopropyl groups; Or its pharmaceutically acceptable salt.

2. The compound or salt according to claim 1, wherein R 1 R 2 R 3 For H.

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

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

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

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

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 compound or salt according to claim 1, wherein the compound is: ， Or its pharmaceutically acceptable salt.

9. The compound or salt according to claim 8, wherein the compound is 。 10. The compound or salt according to claim 8, wherein it is a pharmaceutically acceptable salt of the following compounds. 。 11. A method of treating a patient with diabetic nephropathy, chronic kidney disease, acute kidney injury, hypertension, refractory hypertension, Parkinson's disease, atherosclerotic cardiovascular disease, angina pectoris, myocardial infarction, heart failure, pulmonary hypertension, ischemic stroke, cognitive deficits associated with schizophrenia, cognitive impairment associated with Alzheimer's disease, motor disorders, or attention deficit and hyperactivity disorder, comprising administering to a patient in need an effective amount of any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof.

12. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 10 is used in a therapeutic manner.

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

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

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

16. A method 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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