Pyrrolopyrimidinone derivative and pharmaceutical composition comprising same as active ingredient

Abstract

Disclosed herein are a novel pyrrolopyrimidinone derivative having an excellent PDE5 activity inhibitory effect and pharmaceutical uses thereof. More specifically, a pyrrolopyrimidinone derivative having an excellent PDE5 activity inhibitory effect, an intermediate thereof, a preparation method therefor, and a pharmaceutical composition comprising same as an active ingredient are disclosed. In one aspect, the pyrrolopyrimidinone derivative of the present invention exhibit excellent PDE5 inhibitory ability and pharmacokinetic properties, and thus are expected to provide an excellent therapeutic effect according to PDE5 inhibition in the field of diseases, and thus can be used as an active ingredient of pharmaceuticals.

Description

Pyrrolopyrimidinone derivatives and pharmaceutical compositions containing the same as active ingredients

[0001] The present specification discloses a novel pyrrolopyrimidinone derivative having an excellent inhibitory effect on PDE5 (Phosphodiesterase type 5) overactivity and a pharmaceutical use thereof, and more specifically, a pyrrolopyrimidinone derivative having an excellent inhibitory effect on PDE5 overactivity, an intermediate thereof, a method for manufacturing thereof, and a pharmaceutical composition containing the same as an active ingredient.

[0002] Cross-reference regarding related applications

[0003] This application claims priority to Korean Patent Application No. 10-2024-0194605 filed on December 23, 2024 and Korean Patent Application No. 10-2025-0203552 filed on December 18, 2025, the entire contents of which are incorporated by reference into this application.

[0004]

[0005] Dementia is a progressive degenerative disease of the brain primarily associated with aging. As of 2020, the number of dementia patients worldwide was approximately 55 million, and it is projected to reach about 78 million by 2030 and 139 million by 2050 (World Alzheimer Report 2024). Alzheimer's disease accounts for about 70% of all dementia cases; while it typically occurs after age 65, it can also develop in individuals in their 40s or 50s. This disease is characterized by the gradual decline of mental functions, including memory, thinking ability, judgment, and learning capacity. As the disease progresses, it is accompanied not only by cognitive decline but also by various neuropsychiatric symptoms such as personality changes, agitation, depression, delusions, hallucinations, increased aggression, and sleep disorders. In the late stages, along with neurological disorders such as rigidity and gait abnormalities, physical complications such as fecal and urinary incontinence, infections, and bedsores may also occur.

[0006] Analysis of brain tissue from Alzheimer's disease patients reveals characteristic lesions such as neuritic plaques and neurofibrillary tangles, and macroscopically, overall brain atrophy due to neuronal loss is confirmed. The primary cause of the disease is brain cell damage resulting from the excessive production and deposition of beta-amyloid protein; furthermore, hyperphosphorylation of tau protein—which plays a crucial role in maintaining the skeletal structure of brain cells—along with inflammatory responses and oxidative damage, induce brain cell damage and contribute to the disease. Neuritic plaques, a representative brain pathological finding, are associated with the deposition of beta-amyloid protein, while neurofibrillary tangles are linked to the hyperphosphorylation of tau protein.

[0007] Meanwhile, research results have reported that sildenafil, a phosphodiesterase type 5 (PDE5) inhibitor, has demonstrated clinical efficacy against Alzheimer's disease (Nature Aging, 2021; Neurology, 2024). Additionally, mirodenafil, the only PDE5 inhibitor currently being developed as an oral treatment for Alzheimer's disease, is currently undergoing Phase 3 clinical trials (Alzheimer's Drug Discovery Foundation, 2024). However, mirodenafil requires the maintenance of more stable concentrations within brain tissue through improved blood-brain barrier (BBB) ​​permeability. On the other hand, even with lower permeability, there is a need for higher absolute concentrations within brain tissue to ensure more stable drug efficacy. Therefore, the development of effective treatments for Alzheimer's disease through PDE5 inhibition remains urgently required.

[0008] Accordingly, the inventors, through efforts to develop an effective therapeutic agent, developed a novel pyrrolopyrimidinone derivative that exhibits excellent PDE5 inhibitory effects, improved blood-brain barrier permeability, or higher absolute concentration within brain tissue, thereby presenting the possibility of utilizing this as an oral treatment for Alzheimer's disease and completing the present invention.

[0009]

[0010] One object of the present invention is to provide a pyrrolopyrimidinone derivative having an excellent inhibitory effect on PDE5 (Phosphodiesterase type 5) activity, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof.

[0011] Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of diseases induced by PDE5 activity, comprising the above-mentioned pyrrolopyrimidinone derivative, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof as an active ingredient.

[0012] Another object of the present invention is to provide a method for preparing the above-mentioned pyrrolopyrimidinone derivatives, pharmaceutically acceptable salts thereof, hydrates thereof, or solvates thereof.

[0013] However, the technical problems that the present invention aims to solve are not limited to those mentioned above.

[0014]

[0015] To achieve the above objective, the present invention, in one aspect, provides a pyrrolopyrimidinone derivative represented by Formula 1, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof:

[0016] [Chemical Formula 1]

[0017]

[0018] In the above chemical formula 1,

[0019] R1 is methyl or ethyl;

[0020] R2 is 4-(oxetan-3-yl).

[0021] In another aspect, the present invention provides a composition for the prevention or treatment of diseases induced by PDE5 activity (e.g., neurological brain disease, cerebrovascular disease, sexual dysfunction, cardiovascular disease, hypertension, urinary tract disease, etc.) comprising, as an active ingredient, a pyrrolopyrimidinone derivative represented by Formula 1, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof.

[0022] In another aspect, the present invention provides a method for preparing a pyrrolopyrimidinone derivative represented by Formula 1, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof, comprising the steps as shown in Reaction Scheme 1: (a) combining a compound represented by Formula 2 and a compound represented by Formula 3 to form an amide bond and preparing a compound represented by Formula 4 (Step 1); (b) reacting the compound represented by Formula 4 with a base to produce a compound represented by Formula 5 through a cyclization reaction (Step 2); (c) introducing a sulfonyl chloride group into the compound represented by Formula 5 to prepare a compound represented by Formula 6 (Step 3); and (d) adding a substituted piperazine to the compound represented by Formula 6 to synthesize a compound represented by Formula 1 (Step 4).

[0023] [Reaction Equation 1]

[0024]

[0025] In one aspect, the pyrrolopyrimidinone derivatives of the present invention, their pharmaceutically acceptable salts, their hydrates, or their solvates can effectively inhibit PDE5 (Phosphodiesterase type 5) activity, so they can be used for the prevention and treatment of diseases induced by PDE5 hyperactivity (e.g., neurological brain disease, cerebrovascular disease, sexual dysfunction, cardiovascular disease, hypertension, and urinary tract disease).

[0026] In another aspect, the pyrrolopyrimidinone derivatives, pharmaceutically acceptable salts, and their hydrates or solvates of the present invention can act as PDE5 inhibitors (PDE5i) and exhibit neuroprotective effects by inhibiting beta-amyloid (Aβ) accumulation and reducing neuroinflammation. Furthermore, they can reduce neurofibrillary tangles by inhibiting the hyperphosphorylation of tau protein and improve synaptic plasticity and memory by enhancing CREB signaling through the activation of the cAMP / PKA pathway. Additionally, by inhibiting GSK-3β activity, they can prevent the hyperphosphorylation of tau protein and contribute to cell survival and neuroprotection. Through these various mechanisms, the pyrrolopyrimidinone derivatives and related substances of the present invention are expected to be effective in treating dementia, such as Alzheimer's disease.

[0027] In another aspect, the pyrrolopyrimidinone derivatives of the present invention, their pharmaceutically acceptable salts, their hydrates, or their solvates exhibit increased blood-brain barrier (BBB) ​​permeability and excellent persistence within the brain, thereby exhibiting an enhanced brain distribution profile or having high absolute concentrations within brain tissue, so they can exert pharmacological effects in the brain and be utilized for the prevention or treatment of neurodegenerative diseases (e.g., Alzheimer's disease).

[0028] In another aspect, the pyrrolopyrimidinone derivatives of the present invention, their pharmaceutically acceptable salts, their hydrates, or their solvates have low cytotoxicity, ensuring a sufficient safety margin, and the risk of cardiotoxicity (ECG QT prolongation) is low, so the risk of cardiac-related side effects such as arrhythmia is expected to be significantly lower during clinical use.

[0029]

[0030] Figure 1 compares the PDE5 inhibitory effect and selectivity (PDE6C / PDE5A) of compound 1 and sildenafil.

[0031] Figure 2 compares the PDE5 inhibitory effect and selectivity (PDE6C / PDE5A) of compound 2 and sildenafil.

[0032] Figure 3 compares the BBB permeability of compound 1, compound 2, and mirodenafil.

[0033] Figure 4 shows the results of analyzing the response curve of compound 1 to Phosphodiesterase PDE5A.

[0034] Figure 5 shows the results of analyzing the response curve of compound 1 to Phosphodiesterase PDE6C.

[0035] Figure 6 shows the results of analyzing the response curve of compound 2 to Phosphodiesterase PDE5A.

[0036] Figure 7 shows the results of analyzing the response curve of compound 2 to Phosphodiesterase PDE6C.

[0037] Figure 8 shows the results of analyzing the response curve of sildenafil to Phosphodiesterase PDE5A.

[0038] Figure 9 shows the results of analyzing the response curve of sildenafil to Phosphodiesterase PDE6C.

[0039] Figure 10 shows the results of comparing the plasma concentration-time curves and AUC of Compound 2 and Mirodenafil. Although the systemic exposure of the two drugs was nearly similar, Compound 2 had a half-life (t) compared to Mirodenafil. 1 / 2 It was found that the persistence in the body improved as ) increased by about 27%.

[0040] Figure 11 shows the results of comparing the BBB permeability of Compound 2 and Mirodenafil with brain concentrations over time. It indicates that Compound 2 has superior persistence of brain exposure through the blood-brain barrier (BBB) ​​compared to Mirodenafil.

[0041] Figure 12 shows the results of comparing the BBB permeability of mirodenafil and compound 2 as the brain / plasma concentration ratio (Kp) over time. Compared to mirodenafil, compound 2 showed a brain penetration rate approximately 35% higher and was found to maintain continuous brain exposure for a longer period of time.

[0042] Figure 13 compares plasma and brain concentration-time profiles after a single oral (PO) administration (20 mg / kg) to mice. Vehicle: DMSO: 0.5% CMC (Carboxymethylcellulose) + 0.2% Tween 80 in DDW = 5 : 95.

[0043] Figure 14 shows the polarization graph of E-4031 according to concentration. IC of E-4031 (positive control) 50 Since it was analyzed to be 51 nM (reference range: 10 nM~100 nM), the standardization of the test system was verified.

[0044] Figure 15 shows the polarization graph of mirodenafil according to concentration. As a result of evaluating the hERG channel inhibitory activity of mirodenafil, the IC of mirodenafil 50 It was analyzed to be 9.91 μM.

[0045] Figure 16 shows the polarization graph of E-4031 according to concentration. IC of E-4031 (positive control) 50 Since it was analyzed to be 26 nM (reference range: 10 nM~100 nM), the standardization of the test system was verified.

[0046] Figure 17 shows the polarization graph of Compound 2 at different concentrations. As a result of evaluating the hERG channel inhibitory activity of Compound 2, the IC of Compound 2 50 It was analyzed to be 20.22 μM.

[0047] Fig. 18 is Aβ 1-42 This relates to the administration and experimental schedule for evaluating the efficacy of Compound 2 in a mouse model of cognitive impairment induced by Alzheimer's Disease. AD: Alzheimer's Disease.

[0048] Figure 19 shows the results of the Y-maze test.

[0049] Hereinafter, embodiments and examples of the present invention will be described in detail with reference to the attached drawings so that those skilled in the art can easily implement the present invention. However, the present invention may be embodied in various forms and is not limited to the embodiments and examples described herein.

[0050]

[0051] The present invention will be described in detail below.

[0052]

[0053] In this specification, when a part is described as "comprising" a certain component, this means that, unless specifically stated otherwise, it does not exclude other components but may include additional components.

[0054] In this specification, the term "pharmaceuticalally acceptable salt" refers to any acid-addition salt or base-addition salt that is non-toxic and harmless to the patient and whose side effects caused by said salt do not impair the beneficial efficacy of the compound of the present invention. Inorganic acids that form suitable salts include hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid, hydrobromide, hydroiodide, nitrous acid, or phosphoric acid, and organic acids that form suitable salts include glycolic acid, lactic acid, pyruvate, malonic acid, succinic acid, fumaric acid, malic acid, tartaric acid, citric acid, ascorbic acid, maleic acid, benzoic acid, phenylacetic acid, cinnamic acid, salicylic acid, nicotinic acid, tosylic acid, camphosulfonic acid, naphthoic acid, acetic acid, trifluoroacetic acid, oxalic acid, manderic acid, propionic acid, citric acid, lactic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, carboxylic acid, vanillic acid, benzenesulfonic acid, p-toluenesulfonic acid, or methanesulfonic acid, and preferably oxalates, hydrochlorides, It may be, but is not limited to, benzenesulfonate, hemifumarate, succinate, hemimaleate, nicotinate, tosylate, glycolate, hemisulfonate, tartrate, maleate, aspartate, malate, citrate, malonate, phosphate, glutamate, camphosulfonate, 3-hydroxy-2-naphthoate, mesylate, or 4-hydroxybenzoate. These salts may exist in hydrate, solvate, or substantially anhydrous forms. Additionally, pharmaceutically acceptable metal salts may be prepared using a base, and metal salts include, but are not limited to, sodium, potassium, or calcium salts.

[0055] In this specification, the term "solvent" refers to a solid in which a compound molecule and a solvent molecule form a complex, and "hydrate" refers to a specific solvate in which the solvent is water.

[0056] In this specification, compounds, their pharmaceutically acceptable salts, hydrates, solvates, hydrates of salts, and solvates of salts mean that each includes both crystalline and amorphous forms.

[0057] In this specification, compounds, their pharmaceutically acceptable salts, hydrates, solvates, hydrates of salts, or solvates of salts may be prepared from readily available starting materials using modifications to synthesis protocols known to those skilled in the art.

[0058] In this specification, compound 1 has the following structure:

[0059] .

[0060] In this specification, compound 2 has the following structure:

[0061] .

[0062] In this specification, sildenafil has the following structure:

[0063] .

[0064] In this specification, mirodenafil has the following structure:

[0065] .

[0066] In one aspect, the present invention relates to a pyrrolopyrimidinone derivative represented by Formula 1, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof:

[0067] [Chemical Formula 1]

[0068] .

[0069] In the above chemical formula 1,

[0070] R1 is methyl or ethyl;

[0071] R2 is 4-(oxetan-3-yl).

[0072] In one exemplary embodiment, the pyrrolopyrimidinone derivative may be 5-methyl-2-(5-((4-(oxetan-3-yl)piperazin-1-yl)sulfonyl)-2-propoxyphenyl)-7-propyl-1,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one.

[0073] In one exemplary embodiment, the pyrrolopyrimidinone derivative may be 5-ethyl-2-(5-((4-(oxetan-3-yl)piperazin-1-yl)sulfonyl)-2-propoxyphenyl)-7-propyl-1,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one.

[0074] In another aspect, the present invention relates to a pharmaceutical composition for the prevention or treatment of diseases induced by PDE5 activity, comprising the above-mentioned pyrrolopyrimidinone derivative, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof as an active ingredient.

[0075] In this specification, the term “application” means providing the composition to a subject of application by any appropriate method, and includes administration, application, absorption, and ingestion. In this case, the subject of application refers to any animal, such as a human, monkey, dog, goat, pig, or rat, to which the composition can be applied.

[0076] In this specification, the term “prevention” refers to any act of suppressing or delaying a disease or disorder by applying the above composition, and the term “treatment” refers to any act of improving or beneficially altering the symptoms of an individual suspected of or suffering from a disease or disorder by applying the above composition.

[0077] In an exemplary embodiment, when a compound is administered for therapeutic or preventive purposes, the oral dosage may generally be set in the range of 5 mg to 500 mg per day based on an adult patient (average weight 60 to 70 kg). Accordingly, for adult patients, 5 mg to 500 mg of the active compound per day may be administered in the form of tablets, capsules, or injections in a pharmaceutically acceptable and appropriate formulation, either at once or in multiple doses. In this case, the dosage or formulation may be adjusted according to the patient's age, weight, response, and disease state. The above dosages are based on general cases, and higher or lower dosages may be appropriate depending on the individual case. This is included within the scope of the present invention.

[0078] In an exemplary embodiment, the composition may be characterized as being for oral administration, but is not limited thereto.

[0079] In an exemplary embodiment, the disease induced by the PDE5 activity includes neurological brain disease, cerebrovascular disease, sexual dysfunction, cardiovascular disease, hypertension, and urological disease, and the composition is intended to prevent and treat the various diseases mentioned above by inhibiting PDE5 activity and may be one or more selected from the listed group, but is not limited thereto.

[0080] In an exemplary embodiment, the neurological brain disease includes Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), Huntington's disease, frontotemporal dementia, progressive supranuclear palsy, multiple system atrophy, corticobasal degeneration, dementia with Lewy bodies, Charcot-Marie-Tooth disease, and prion diseases, and the composition is intended to improve, prevent, and treat diseases associated with the accumulation of various beta-amyloid and tau proteins, and may be one or more selected from the listed group, but is not limited thereto.

[0081] In an exemplary embodiment, the cerebrovascular disease includes ischemic stroke, hemorrhagic stroke, cerebral small vessel disease, cerebral vasospasm, and vascular dementia, and the composition is intended to prevent and treat these various cerebrovascular diseases and may be one or more selected from the listed group, but is not limited thereto.

[0082] In an exemplary embodiment, the sexual dysfunction includes erectile dysfunction, sexual arousal disorder, premature ejaculation, and decreased sexual satisfaction, and the composition is intended to improve, prevent, and treat these various sexual dysfunctions and may be one or more selected from the listed group, but is not limited thereto.

[0083] In an exemplary embodiment, the cardiovascular disease includes heart failure, cardiac hypertrophy, coronary artery disease, atherosclerosis, peripheral arterial disease, and single ventricular heart Fontan operation, and the composition is intended to prevent and treat these various cardiovascular diseases and may be one or more selected from the listed group, but is not limited thereto.

[0084] In an exemplary embodiment, the hypertension includes pulmonary arterial hypertension, essential hypertension, hypertensive heart disease, and renovascular hypertension, and the composition is intended to prevent and treat these various types of hypertension and may be one or more selected from the listed group, but is not limited thereto.

[0085] In an exemplary embodiment, the urinary tract disease includes benign prostatic hyperplasia, and the benign prostatic hyperplasia includes, but is not limited to, benign prostatic hyperplasia accompanied by lower urinary tract symptoms (LUTS).

[0086] In an exemplary embodiment, the pharmaceutical composition may be provided in any formulation suitable for topical application. For example, it may be administered orally, transdermally, intravenously, intramuscularly, or subcutaneously. As an example, the pharmaceutical composition may be an injectable, a topical solution, a suspension, an emulsion, a gel, a patch, or a spray, but is not limited thereto. The formulation may be readily prepared according to conventional methods in the art, and surfactants, excipients, wettable powders, emulsification promoters, suspensions, salts or buffers for osmotic pressure regulation, coloring agents, flavorings, stabilizers, preservatives, preservatives, or other commonly used adjuvants may be appropriately used.

[0087] In an exemplary embodiment, the active ingredient of the pharmaceutical composition will vary depending on the subject's age, gender, weight, pathological condition and its severity, route of administration, or the prescriber's judgment. Determining the appropriate dosage based on these factors is within the level of those skilled in the art.

[0088] In an exemplary embodiment, the pharmaceutical composition may be administered in an effective amount to a subject requiring treatment, prevention, or symptom relief. The subject may include human or non-human animals, and non-human animals may include, but are not limited to, mammals, birds, reptiles, or amphibians. Examples of companion animals may include dogs, cats, rabbits, ferrets, guinea pigs, hamsters, birds (e.g., parrots), and examples of livestock may include cattle, horses, pigs, sheep, goats, poultry, etc.

[0089] In an exemplary embodiment, the pharmaceutical composition may be applied for the treatment, prevention, or improvement of cognitive impairment in companion animals.

[0090] In this specification, the terms “cognitive dysfunction” or “Cognitive Dysfunction Syndrome (CDS)” refer to a chronic, progressive neurodegenerative condition observed primarily in middle-aged and elderly companion animals, characterized by a decline in cognitive domains such as learning, memory, spatial orientation, and social interaction, and accompanying behavioral changes.

[0091] In another aspect, the present invention relates to a method for preventing or treating a disease induced by PDE5 activity by applying a composition comprising the above compound, its salt, its stereoisomer, its solvate, or its hydrate as an active ingredient to a subject.

[0092] In another aspect, the present invention relates to the use of said compound, its salt, its stereoisomer, its solvate, or its hydrate for the preparation of a composition for the prevention or treatment of diseases induced by PDE5 activity.

[0093] In another aspect, the present invention relates to the use of the said compound, its salt, its stereoisomer, its solvate, or its hydrate for the prevention or treatment of diseases induced by PDE5 activity.

[0094] In another aspect, the present invention relates to a method for preparing a pyrrolopyrimidinone derivative, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof, comprising, as shown in Reaction Scheme 1: (a) combining a compound represented by Formula 2 and a compound represented by Formula 3 to form an amide bond and preparing a compound represented by Formula 4 (Step 1); (b) reacting the compound represented by Formula 4 with a base to produce a compound represented by Formula 5 through a cyclization reaction (Step 2); (c) introducing a sulfonyl chloride group into the compound represented by Formula 5 to prepare a compound represented by Formula 6 (Step 3); and (d) adding a substituted piperazine to the compound represented by Formula 6 to synthesize a compound represented by Formula 1 (Step 4).

[0095] [Reaction Equation 1]

[0096] .

[0097] In an exemplary embodiment, in step 1, EDC·HCl (1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide) or DCC (Dicyclohexylcarbodiimide), preferably EDC·HCl, may be used, but is not limited thereto. EDC·HCl has the characteristic of being soluble in water and can effectively reduce side reactions when combined with activators such as HOBt (1-Hydroxybenzotriazole), HOAt (1-Hydroxy-7-azabenzotriazole), and Oxyma (Ethyl cyano(hydroxyimino)acetate).

[0098] In an exemplary embodiment, a base such as t-BuOK, NaH, or K₂CO₃ may be used in step 2, and preferred reaction conditions (e.g., temperature, solvent, etc.) may be appropriately adjusted according to the characteristics of the base and reactants used.

[0099] In an exemplary embodiment, chlorosulfonic acid (ClSO₃H) may be used in step 3, but is not limited thereto.

[0100]

[0101] The structure and effects of the present invention will be explained in more detail below with reference to examples. However, the following examples are provided for illustrative purposes only to aid in understanding the present invention, and the scope and range of the present invention are not limited by them.

[0102]

[0103] [Example]

[0104] [Example 1]

[0105] Preparation of a novel pyrrolopyrimidinone derivative with excellent PDE5 inhibitory effect

[0106] A novel pyrrolopyrimidinone derivative was prepared according to the following reaction scheme 2.

[0107] [Reaction Equation 2]

[0108]

[0109] Experimental Example 1

[0110] Synthesis of 5-methyl-2-(5-((4-(oxetan-3-yl)piperazin-1-yl)sulfonyl)-2-propoxyphenyl)-7-propyl-1,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one

[0111] (1) Step 1: Synthesis of 1-methyl-3-(2-propoxybenzamido)-4-propyl-1H-pyrrole-2-carboxamide

[0112] 2-propoxybenzoic acid (19.64 mmol), HOBT (29.5 mmol), TEA (58.9 mmol), and 3-amino-1-ethyl-4-propyl-1H-pyrrole-2-carboxamide (19.64 mmol) were added sequentially to DCM (40 ml), after which EDC.HCl (29.5 mmol) was slowly added to the mixture over 5 minutes. The reaction mixture was stirred at room temperature for 16 hours. After the reaction was complete, the reaction mixture was diluted with DCM (40 ml) and washed twice with 1N HCl solution. The organic layer was dried with MgSO4, filtered, and the filtered solution was concentrated under reduced pressure. The product obtained by concentration was purified by silica gel flash column chromatography using a mixed solution of DCM and MeOH to obtain 1-methyl-3-(2-propoxybenzamido)-4-propyl-1H-pyrrole-2-carboxamide (yield: 89%, 17.48 mmol).

[0113] (2) Step 2: Synthesis of 5-methyl-2-(2-propoxyphenyl)-7-propyl-1,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one

[0114] A solution of the compound obtained in Step 1 (16.59 mmol) and tert-butoxide (33.19 mmol) suspended in tert-BuOH (60 mL) was reacted at 70°C for 4 hours, after which the reaction mixture was cooled to room temperature and the tert-BuOH was distilled under reduced pressure. The product was diluted with water and extracted with EtOAc. The extract was dried with MgSO4 and filtered, and the filtered solution was concentrated under reduced pressure. The residue was dried under vacuum to obtain 5-methyl-2-(2-propoxyphenyl)-7-propyl-1,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one (yield: 100%, 16.59 mmol).

[0115] (3) Step 3: Synthesis of 3-(5-methyl-4-oxo-7-propyl-4,5-dihydro-1H-pyrrolo[3,2-d]pyrimidin-2-yl)-4-propoxybenzenesulfonyl chloride

[0116] chlorosulfonic acid (3 mL) was slowly added dropwise at 0 °C to a solution in which the compound (4.42 mmol) obtained in Step 2 above was dissolved in DCM (2 mL). The reaction mixture was stirred at room temperature for 1 hour, then treated with ice water, and extracted three times with DCM.

[0117] The combined extract was dried with MgSO4 and filtered. The residue was dried under vacuum to obtain 3-(5-methyl-4-oxo-7-propyl-4,5-dihydro-1H-pyrrolo[3,2-d]pyrimidin-2-yl)-4-propoxybenzenesulfonyl chloride (yield: 59%, 2.61 mmol).

[0118] (4) Step 4: Synthesis of 5-methyl-2-(5-((4-(oxetan-3-yl)piperazin-1-yl)sulfonyl)-2-propoxyphenyl)-7-propyl-1,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one

[0119] TEA (1.89 mmol) and an amine (0.99 mmol) to be introduced as a substituent were added to a solution in which the compound (0.94 mmol) obtained in Step 3 above was dissolved in DCM (10 ml). The reaction mixture was stirred for 1 hour, diluted with DCM, and then washed with water. The organic layer was dried with MgSO4 and filtered. The residue was dried under vacuum, and the product was purified by silica gel flash column chromatography using a mixed solution of DCM and ETOH to obtain 5-methyl-2-(5-((4-(oxetan-3-yl)piperazin-1-yl)sulfonyl)-2-propoxyphenyl)-7-propyl-1,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one (yield: 67%, 0.42 mmol), represented as Compound 1 below.

[0120] [Compound 1]

[0121]

[0122] 1H NMR (400 MHz, CDCl3) 10.69 (br. s., 1H), 8.87 (d, J = 2.50 Hz, 1H), 7.81 (dd, J = 2.44, 8.69 Hz, 1H), 7.15 (d, J = 8.75 Hz, 1H), 6.89 (s, 1H), 4.63 (t, J = 6.57 Hz, 2H), 4.48 (t, J = 6.13 Hz, 2H), 4.25 (t, J = 6.44 Hz, 2H), 4.08 (s, 3H), 3.49 (td, J = 6.30, 12.54 Hz, 1H), 3.14 (br. s., 4H), 2.71 (t, J = 7.38 Hz, 2H), 2.28 - 2.54 (m, 4H), 1.95 - 2.14 (m, 2H), 1.69 - 1.80 (m, 3H), 1.19 (t, J = 7.44 Hz, 3H), 0.99 (t, J = 7.38 Hz, 3H).

[0123]

[0124] Experimental Example 2

[0125] Synthesis of 5-ethyl-2-(5-((4-(oxetan-3-yl)piperazin-1-yl)sulfonyl)-2-propoxyphenyl)-7-propyl-1,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one

[0126] (1) Step 1: Synthesis of 1-ethyl-3-(2-propoxybenzamido)-4-propyl-1H-pyrrole-2-carboxamide

[0127] 1-ethyl-3-(2-propoxybenzamido)-4-propyl-1H-pyrrole-2-carboxamide (yield: 87%) was obtained by preparing it in the same manner as Step 1 of Experimental Example 1 above.

[0128] (2) Step 2: Synthesis of 5-ethyl-2-(2-propoxyphenyl)-7-propyl-1,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one

[0129] 5-ethyl-2-(2-propoxyphenyl)-7-propyl-1,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one (yield: 97%) was obtained by preparing it in the same manner as Step 2 of Experimental Example 1 above.

[0130] (3) Step 3: Synthesis of 3-(5-ethyl-4-oxo-7-propyl-4,5-dihydro-1H-pyrrolo[3,2-d]pyrimidin-2-yl)-4-propoxybenzenesulfonyl chloride

[0131] 3-(5-ethyl-4-oxo-7-propyl-4,5-dihydro-1H-pyrrolo[3,2-d]pyrimidin-2-yl)-4-propoxybenzenesulfonyl chloride was obtained by preparing it in the same manner as Step 3 of Experimental Example 1 above.

[0132] (4) Step 4: Synthesis of 5-ethyl-2-(5-((4-(oxetan-3-yl)piperazin-1-yl)sulfonyl)-2-propoxyphenyl)-7-propyl-1,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one

[0133]

[0134] 5-ethyl-2-(5-((4-(oxetan-3-yl)piperazin-1-yl)sulfonyl)-2-propoxyphenyl)-7-propyl-1,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one (Compound 2, yield: 75%) represented by the following compound 2 was obtained by preparing in the same manner as Step 4 of Experimental Example 1 above.

[0135] [Compound 2]

[0136]

[0137] 1H NMR (400 MHz, CDCl3) 10.71 (br. s., 1H), 8.90 (d, J = 2.38 Hz, 1H), 7.81 (dd, J = 2.50, 8.75 Hz, 1H), 7.15 (d, J = 8.75 Hz, 1H), 6.97 (s, 1H), 4.63 (t, J = 6.57 Hz, 2H), 4.32 - 4.55 (m, 4H), 4.25 (t, J = 6.44 Hz, 2H), 3.49 (quin, J = 6.28 Hz, 1H), 3.14 (br. s., 4H), 2.72 (t, J = 7.44 Hz, 2H), 2.28 - 2.56 (m, 4H), 2.05 (qd, J = 7.04, 13.91 Hz, 2H), 1.68 - 1.80 (m, 2H), 1.48 (t, J = 7.25 Hz, 3H), 1.20 (t, J = 7.44 Hz, 3H), 1.00 (t, J = 7.38 Hz, 3H).

[0138]

[0139] [Example 2]

[0140] PDE5 Inhibitory Activity and PDE6C / PDE5A Selectivity of Novel Pyrrolopyrimidinone Derivatives

[0141] Sildenafil (Viagra®) is well known as a selective PDE5A inhibitor and was first launched as a treatment for erectile dysfunction after receiving approval from the U.S. FDA in 1998. Sildenafil increases intracellular cGMP concentration by competitively binding to the active site of PDE5A and inhibiting the degradation of cGMP. The increased cGMP activates protein kinase G (PKG), which reduces calcium concentration within smooth muscle cells, leading to relaxation of vascular smooth muscle and increased blood flow.

[0142] However, while sildenafil is selective for PDE5A, it also exhibits a certain level of inhibitory activity against the structurally similar PDE6. Since PDE5 inhibitors target sites structurally similar to PDE6C, there is a possibility of simultaneous inhibition of PDE6C; as PDE6C is involved in the light transmission processes of retinal rods and cones, its inhibition may cause temporary visual impairments (such as bluish-green vision or blurred vision). Additionally, sildenafil is known to exhibit cross-reactivity with PDE11.

[0143] The selectivity of PDE5A and PDE6C indicates which of the two enzymes a drug acts more specifically on, and this represents the IC50 for each enzyme. 50 Ratio of values ​​(drug concentration required for 50% enzyme inhibition) (IC 50 PDE6C / IC 50It is calculated as PDE5A. The higher this selectivity ratio, the greater the selectivity for PDE5A, which lowers the risk of visual side effects while maintaining therapeutic efficacy; conversely, the lower the ratio, the greater the likelihood of side effects associated with PDE6C inhibition. Therefore, this selectivity ratio serves as an important indicator for evaluating drug efficacy and safety, and there is a demand for the development of PDE5 inhibitors that possess high selectivity for PDE5A while exhibiting low cross-reactivity with PDE6C and other PDE subtypes.

[0144]

[0145] Experimental Example

[0146] In this experimental example, efficacy and selectivity experiments were conducted using compound 1 and sildenafil as test substances (Experiment 1), and additionally, efficacy and selectivity experiments were conducted using compound 2 and sildenafil as test substances (Experiment 2).

[0147] The inhibitory effects of each test substance (compound 1, compound 2, and sildenafil) on human-derived PDE5A and PDE6C enzymes were evaluated.

[0148] The method for measuring the enzyme inhibitory activity of the test compound and calculating the IC50 is as follows:

[0149] The enzyme inhibitory activity of the compounds of the present invention was measured using two different conditions and methods. After treating each compound at various concentrations, the inhibition rate against the enzyme was measured.

[0150] 1. Experimental Conditions 1 and IC50 Calculation (PDE5A1 and PDE6C Inhibitory Activity)

[0151] Experiment 1 was conducted at Eurofins Panlabs Discovery Services Taiwan (New Taipei City, Taiwan), a specialized testing agency, and all measurements were duplicated. PDE5A and PDE6C were derived from human recombinant insect Sf9 cells, the substrate was fluorescein-labeled cyclic guanosine monophosphate (FAM-cGMP), and the assay was the Fluorescein-GMP-IMAP Fluorescence Polarization (FP) method. The IMAP (Immobilized Metal Affinity for Phosphate) method is a technique that captures and binds phosphate groups released during the reaction between an enzyme, substrate, and inhibitor to trivalent metal ions; the bound complex exhibits high FP, and FP is proportional to PDE5A1 activity. IC50 values ​​were calculated based on the measured enzyme inhibition rate data. The IC50 value was calculated through non-linear least squares regression analysis using MathIQ software (ID Business Solutions Ltd., UK).

[0152] 2. Second Experimental Conditions and IC50 Calculation (PDE5A1 and PDE6C Inhibitory Activity)

[0153] For the second experiment, assay kits from BPS Bioscience (San Diego, CA, USA), specifically human PDE5A1 (Cat. # 60350) and human PDE6C (Cat. # 79501) kits, were used. The source of PDE5A1 and PDE6C was human GST-tag recombinant, the substrate was FAM-cGMP, and the assay method was Fluorescence Polarization (FP). The excitation wavelength was 470 nm, and the emission wavelength was 528 nm. Phosphate groups released from the reaction of the enzyme, substrate, and inhibitor are recognized by specific phosphate nanobeads (binders) to form large complexes (high FP) with restricted movement. FP is proportional to PDE5A1 activity. All measurements were duplicated twice, identical to the first experiment. IC50 values ​​were calculated based on the measured concentration-response data. Data analysis was performed using GraphPad Prism software (GraphPad Software, USA). For the analysis, the IC50 value was determined using a 4-parameter logistic (4PL) nonlinear regression model of the response (% inhibition) curve relative to the log dose (inhibitor concentration).

[0154] 3. Verification of test method objectivity and use of control drugs

[0155] The difference in sensitivity of the IC50 values ​​derived from the two test methods above is attributed to differences in reaction conditions, such as enzyme concentration, substrate concentration, and the type and method of binder used in each test system. This is a phenomenon that occurs in general biochemical analysis methods. To confirm and verify the objectivity and reliability of this test method, the activity was measured under the same conditions using sildenafil, a known compound, as a control drug.

[0156]

[0157] As a result of the first experiment, as shown in FIGS. 1, FIGS. 4, and Table 1, Compound 1 of the present invention has an IC with respect to PDE5A. 50 It exhibited potent inhibitory activity with a value of 0.42 nM. This is sildenafil (IC2). 50 = 0.83 nM) is an inhibitory effect that is about twice as good.

[0158] Inhibitory effects of Compound 1 and Sildenafil on PDE5A and PDE6C enzymes Test Substance SpeciesConc.%InhIC 50 * PDE5A Compound 1 Human 1 nM 690.42 nM Sildenafil Human 1 nM 570.83 nMP DE6C Compound 1 Human 2.56 nM 516.25 nM Sildenafil Human 16 nM 549.94 nM

[0159] * The standard error of the mean is provided from results based on multiple independent measurements.

[0160] As a result of the second experiment, as shown in FIGS. 2, FIGS. 6, and Table 2, Compound 2 of the present invention has an IC with respect to PDE5A. 50 It exhibited potent inhibitory activity with a value of 0.52 nM. This is sildenafil (IC2). 50 = 5.25 nM) corresponds to an inhibitory effect that is more than 10 times superior.

[0161] Inhibitory effects of Compound 2 and Sildenafil on PDE5A and PDE6C enzymes Test Substance SpeciesConc.%InhIC 50 * PDE5A Compound 2Human1 nM640.52 nM Sildenafil Human5 nM515.25 nMP DE6C Compound 2Human16 nM5216.96 nM Sildenafil Human16 nM4417.31 nM

[0162] * The standard error of the mean is provided from results based on multiple independent measurements.

[0163] In addition, the results of the first experiment showed that compound 1 had a PDE6C / PDE5A selectivity ratio of approximately 15 (6.25 nM / 0.42 nM), which is about 1.25 times higher than the selectivity ratio of sildenafil, which is approximately 12 (9.94 nM / 0.83 nM) (Table 1 and Figure 1).

[0164] As a result of the second experiment, the PDE6C / PDE5A selectivity ratio of this compound 2 was calculated to be approximately 32.6 (16.96 nM / 0.52 nM), which is about 10 times higher than the selectivity ratio of sildenafil, which is approximately 3.3 (17.31 nM / 5.25 nM) (Table 2 and Figure 2).

[0165] This means that compound 2 acts very selectively on PDE5A, while minimizing PDE6C inhibition associated with visual side effects.

[0166] Therefore, Compound 2 was identified as the best compound among those tested in terms of PDE6C / PDE5A selectivity and efficacy, and is expected to provide enhanced efficacy and safety compared to existing PDE5 inhibitors as a treatment for central nervous system diseases, including Alzheimer's disease. Some subsequent experiments were conducted on Compound 2, the best compound.

[0167]

[0168] [Example 3]

[0169] Excellent brain distribution and persistence of novel pyrrolopyrimidinone derivatives

[0170] Experimental Example 1

[0171] Comparison of Blood-Brain Barrier (BBB) ​​Permeability and Brain Distribution

[0172] Mirodenafil (trade name: Mvix®) is a selective PDE5A inhibitor developed in Korea and was launched as a treatment for erectile dysfunction after receiving approval from the Ministry of Food and Drug Safety in Korea in 2007. Mirodenafil has a mechanism of action similar to sildenafil and induces vascular smooth muscle relaxation and increased blood flow by competitively binding to the active site of PDE5A and inhibiting the degradation of cGMP.

[0173] In this experiment, Compound 1, Compound 2, and Mirodenafil were orally administered to ICR mice at a dose of 10 mg / kg each. The test substances were prepared at a concentration of 2 mg / mL and administered at a volume of 5 mL / kg, using a mixture of 10% DMSO, 10% Cremophor EL, 40% PEG400, and 40% DDW as the solvent. Plasma and tissues (liver, lung, and brain) were collected 30 minutes after administration, and tissue distribution was analyzed (n=3).

[0174] The results of the tissue distribution analysis of each test substance are as shown in Tables 3 to 5 below.

[0175] Compound 1 Tissue Distribution Analysis (PO, 10 mg / kg, n=3, at 30 minutes) Analysis variablesTissueMouse1Mouse2Mouse3MEANSDTissue concentration(ng / g tissue)Liver7720.006640.002508.005622.6672750.902Lung3856.003884.00424.002721.3331989.598Brain68.4095.6032.4465.48031.681Plasma concentrationat 30 min (ng / mL)1360.001390.00784.001178.000341.544TissueMouse1Mouse2Mouse3MEANSDKp ratio (30 min)(Tissue to plasma concentration ratio) Liver5.6764.7773.1994.5511.254Lung2.8352.7940.5412.0571.313Brain0.0500.0690.0410.0530.014

[0176] * LOQ: 1 ng / mL

[0177] As shown in Table 3, tissue distribution studies at 30 minutes after oral administration of Compound 1 (10 mg / kg) revealed an average concentration of 65.48 ng / g (SD: ±31.681) in the brain, and K in the brain p The ratio averaged 0.053 (SD: ±0.014).

[0178] Compound 2 Tissue Distribution Analysis (PO, 10 mg / kg, n=3, at 30 minutes) Analysis variablesTissueMouse1Mouse2Mouse3MEANSDTissue concentration(ng / g tissue)Liver4400.004760.003448.004202.667677.895Lung600.001620.001424.001214.667541.263Brain49.2028.1659.2045.52015.844Plasma concentrationat 30 min (ng / mL)332688656558.667196.950TissueMouse1Mouse2Mouse3MEANSDKp ratio (30 min)(Tissue to plasma concentration ratio) Liver13.2536.9195.2568.4764.220Lung1.8072.3552.1712.1110.279Brain0.1480.0410.0900.0930.054

[0179] * LOQ: 1 ng / mL

[0180] As shown in Table 4, tissue distribution studies at 30 minutes after oral administration (10 mg / kg) of Compound 2 revealed that it was detected in the brain at an average concentration of 45.520 ng / g (SD: ±15.844), and K in the brain p The ratio averaged 0.093 (SD: ±0.054).

[0181] Mirodenafil tissue distribution analysis (PO, 10 mg / kg, n=3, at 30 minutes)Analysis variablesTissueMouse1Mouse2Mouse3MEANSDTissue concentration(ng / g tissue)Liver1932.003344.003952.003076.0001036.324Lung1284.001276.002088.001549.333466.516Brain28.0822.9237.8029.6007.556Plasma concentrationat 30 min (ng / mL)378.00343.00493.00404.66778.475TissueMouse1Mouse2Mouse3MEANSDKp ratio (30 min)(Tissue to plasma concentration ratio) Liver5.1119.7498.0167.6262.344Lung3.3973.7204.2353.7840.423Brain0.0740.0670.0770.0730.005

[0182] * LOQ: 1 ng / mL

[0183] As shown in Table 5, tissue distribution studies at 30 minutes after oral administration of mirodenafil (10 mg / kg) revealed an average concentration of 29.6 ng / g brain (SD: ±7.556), and K in the brain p The ratio is an average of 0.073 (SD: ±0.005).

[0184] Synthesizing the experimental results above, as shown in Figures 11, 13, and Table 6, it was confirmed that Compound 1 and Compound 2 effectively pass through the blood-brain barrier (BBB) ​​and are distributed in brain tissue upon oral administration. For Compound 1, the brain concentration was measured at 65.48 ng / g brain 30 minutes after oral administration of 10 mg / kg, indicating a brain concentration approximately 2.21 times higher than that of Mirodenafil (29.6 ng / g brain). For Compound 2, the brain concentration was 45.52 ng / g brain 30 minutes after oral administration of 10 mg / kg, which was higher than that of Mirodenafil (29.6 ng / g brain); furthermore, the Kp,brain value (0.093) was also higher than that of Mirodenafil (0.073), demonstrating superior characteristics in terms of BBB penetration and brain distribution.

[0185] Comparison of Brain Distribution and BBB Permeation by Test Substance (10 mg / kg, 30 min) Test Substance Brain Concentration (ng / g brain) Kp, brain (Brain / Plasma Ratio) Compound 165.48 (ng / g brain) 0.053 Compound 245.52 (ng / g brain) 0.093 Mirodenafil 29.6 (ng / g brain) 0.073

[0186]

[0187] Experimental Example 2

[0188] To evaluate the persistence of brain distribution of compound 2 and mirodenafil, they were orally administered to ICR mice at a dose of 20 mg / kg each, and plasma and brain tissues were collected at 0, 30, 60, and 120 minutes to measure drug concentrations (n=3).

[0189] As a result, as shown in Tables 7 to 12 below, Mirodenafil showed a high peak in average plasma concentration at 30 minutes after administration, but subsequently decreased rapidly. On the other hand, Compound 2 had a relatively low initial average plasma concentration of 1,438 ng / mL, but maintained a higher concentration than Mirodenafil even at 120 minutes, demonstrating superior persistence. In other words, it exhibited a drug-friendly PK profile that enhanced blood persistence and effective concentration maintenance while lowering the risk of peak side effects compared to Mirodenafil.

[0190] Regarding brain tissue concentrations, mirodenafil showed a relatively high peak with an average concentration of 155 ng / g brain at the 30-minute mark, but subsequently decreased rapidly. On the other hand, although the initial brain concentration of Compound 2 was somewhat lower at 83 ng / g brain, it reached a level (72 ng / g brain) similar to mirodenafil (78 ng / g brain) at the 60-minute mark and maintained a higher concentration than mirodenafil at the 120-minute mark, demonstrating significantly superior persistence within the brain. Considering that sustained drug exposure is key to the manifestation of effect in the regulation of the CREBpTauAβ pathway through PDE5 inhibition, these results suggest that Compound 2 possesses a mechanistically more favorable brain exposure pattern compared to mirodenafil.

[0191] In particular, regarding the brain / plasma concentration ratio (Kp,brain), Compound 2 consistently showed high values ​​over time (30 min: 0.063, 60 min: 0.080, 120 min: 0.064). On the other hand, Mirodenafil showed lower Kp,brain values ​​than Compound 2 overall (30 min: 0.045, 60 min: 0.054, 120 min: 0.064). In summary, Compound 2 has a higher distribution efficiency to the brain relative to the blood starting from the 30-minute mark, and at 60 minutes, it clearly outperforms Mirodenafil in drug retention within the brain. Furthermore, brain concentrations remain stable up to 120 minutes, and the balance between brain and blood concentrations is maintained gradually, confirming that substantial brain exposure is secured for an extended period.

[0192] Plasma concentration of Compound 2 (PO, 20 mg / kg) Time(min)#1#2#3#4meanS.D.000000030194062236828201438115160940980117080697415012022739411901280773539

[0193] Brain concentration of Compound 2 (PO, 20 mg / kg) Time(min)#1#2#3#4meanS.D.000000030140.857.616.81188357607034.248.8135.6724512015.7247.258.819.923521

[0194] Comparison of brain / plasma concentration ratio of Compound 2 (PO, 20 mg / kg) Time(min)#1#2#3#4meanS.D.0N / AN / AN / AN / A300.0730.0930.0460.0420.0630. 024600.0740.0350.0420.1680.0800.0611200.0690.1200.0490.0160.0640.044

[0195] N / A: not applicable, interference observed, analysis not possible

[0196] Plasma concentration of mirodenafil (PO, 20 mg / kg)Time(min)#1#2#3#4#5meanS.D.00000000305700305557301575301038141835601580180011201510150328312022627020223334

[0197] Brain concentration of mirodenafil (PO, 20 mg / kg) Time(min)#1#2#3#4#5meanS.D.0000000030173.2134227.297.2144.4155496071.684.491.663.6781312010.1226.2410.4169

[0198] Comparison of brain / plasma concentration ratios of mirodenafil (PO, 20 mg / kg) Time(min)#1#2#3#4#5meanS.D.0N / AN / AN / AN / AN / A300.0300.0440.0400.0620.048 0.0450.012600.0450.0470.0820.0420.0540.0191200.0450.0970.0510.0640.029

[0199] N / A: not applicable, interference observed, analysis not possible

[0200]

[0201] As shown in Figures 10, 12 and Table 13 below, Compound 2 is maintained in the blood-brain for a longer period compared to mirodenafil (average Kp +35%), and has a brain concentration at 120 minutes that is more than twice as high, possessing a PKBBB profile optimized for sustained CNS therapeutic effects. In other words, unlike peak-dependent mirodenafil, Compound 2 is a structurally superior CNS therapeutic candidate that simultaneously possesses stable exposure, excellent BBB permeability, and high brain retention.

[0202] Comparison of Kp and brain over time for Compound 2 and Mirodenafil Item Compound 2 (Mean ± SD, n=4) Mirodenafil (Mean ± SD, n=3-5) 30 min 0.063 ± 0.02 40.045 ± 0.01 260 min 0.080 ± 0.06 10.054 ± 0.01 91 20 min 0.064 ± 0.04 40.064 ± 0.02 9 AUC brain / AUC plasma 0.0618(6808.05 / 110137.5)0.0456(8624.6 / 189012.5)

[0203]

[0204] [Example 4]

[0205] Evaluation of Pharmacokinetic (PK) Characteristics of Novel Pyrrolopyrimidinone Derivatives

[0206] Experimental Example 1

[0207] Evaluation of drug persistence in the body

[0208] To evaluate the drug persistence of compound 2 and mirodenafil in the body, they were orally administered to ICR mice at a dose of 20 mg / kg each, and plasma was collected at 15, 30, 60, 120, 240, and 480 minutes to measure drug concentrations (n=3 or 4).

[0209] The results are as described in Tables 14 and 15. Mirodenafil had a very high initial peak (C) with an average plasma concentration of 4.133 μg / mL at 15 minutes after administration. max It showed a rapid decrease afterward. On the other hand, although the initial average concentration of Compound 2 was low at 1.264 μg / mL at the 15-minute mark, it showed superior persistence by maintaining a higher concentration than Mirodenafil even at 60 minutes (1.363 μg / mL) and 120 minutes (0.687 μg / mL). Therefore, it was confirmed that Compound 2 provides longer-term drug exposure with a lower risk of adverse reactions due to its low initial peak.

[0210] Plasma concentration of Compound 2 (PO, 20 mg / kg) Time(min)#1#2#3meanS.D.150.2332.4401.1201.2641.111300.4621.8701.8701.4010.813600.6942.0251.370 1.3630.6661200.6631.0800.3180.6870.3822400.1350.1210.0370.0980.0534800.0020.0070.0080.0050.003

[0211] Plasma concentration of mirodenafil (PO, 20 mg / kg) Time(min)#1#2#3#4meanS.D.153.9554.5304.0753.9704.1330.270302.8902.3052.1552.1002.3630.362601.4951.1900.45 90.8921.0090.4421200.4640.7540.0380.1730.3570.3192400.0810.0280.0080.0210.0350.0324800.010BQLBQLBQL0.010-

[0212] In addition, as shown in Table 16, the PK parameter analysis results indicated that the Cmax of mirodenafil was 4.133 μg / mL and that of Compound 2 was 1.668 μg / mL, exhibiting a peak approximately 2.5 times lower than that of mirodenafil. However, the AUC of the two substances was nearly equivalent (AUC 95.8%), at 198.4 µg / mL*min and 192.3 µg / mL*min, respectively, and the half-life (t) of Compound 2 1 / 2 ) showed a longer systemic duration of 50.4 minutes, which is about 25% higher than mirodenafil (39.8 minutes).

[0213] In addition, the time to reach the maximum concentration of compound 2 (t max The duration was 35 minutes, which is about 2.3 times longer than that of mirodenafil (15 minutes). This means that the onset of action proceeds more gradually and stably without forming a sharp peak, providing a favorable profile in terms of sustained action. Additionally, the mean retention time (MRT) of compound 2 was 91.6 minutes, which is about 1.8 times longer than that of mirodenafil (50.3 minutes), indicating that the drug remains stably in the body for a longer period.

[0214] That is, compound 2 is C max ga is lower, and t maxIt possesses unique pharmacokinetic characteristics, with a significantly longer MRT compared to mirodenafil while maintaining an equivalent AUC. This profile enables stable and sustained drug exposure in the body without abrupt peaks, consequently providing a competitive edge as a safer and longer-acting PDE5 inhibitor candidate.

[0215] Compound 2, Mirodenafil mouse PO PK parameters (suspension, 20 mg / kg) PK parameters Compound 2 Mirodenafil Mean ± SD(n=3) Mean ± SD(n=4) C max (ug / mL)1.668±0.8904.133±0.270t max (min)35.0±22.9(15-60)15.0±0.0(15)AUC last (ug / mL*min)191.9±85.7197.5±57.3AUC inf (ug / mL*min)192.3±85.9198.4±57.7t 1 / 2 (min)50.4±8.139.8±17.8MRT (min)91.6±25.050.3±15.7

[0216]

[0217] [Example 5]

[0218] hERG Cardiac Safety Assessment and Safety Margin Comparison

[0219] hERG (Human Ether-a-go-go-Related Gene) potassium channels play a crucial role in the cardiac repolarization process, and inhibition of these channels can cause severe cardiotoxicity, such as QT interval prolongation and ventricular arrhythmias (Torsades de Pointes). Therefore, the potential for hERG channel inhibition is utilized as a key indicator for evaluating cardiac safety during the drug development process, and hERG IC5 50 Value and maximum plasma concentration (C max The larger the safety margin calculated as the ratio between ), the lower the risk of cardiotoxicity in clinical practice is assessed.

[0220] In this experiment, we aimed to evaluate the hERG channel inhibitory activity of compound 2 and mirodenafil and confirm the superiority of compound 2 in cardiac safety by comparing the safety margins based on mouse pharmacokinetic data.

[0221]

[0222] Experimental Example 1

[0223] Evaluation of the hERG inhibitory activity of mirodenafil

[0224] The inhibitory activity of mirodenafil on the Human Ether-a-go-go-Related Gene (hERG) potassium channel was measured using a non-cell based fluorescence polarization (FP) assay method.

[0225] The experiment was performed as follows: 1) E-4031 (positive control) and the test substance were serially diluted (dilution factor: 3, 16 doses) and dispensed into 384 wells, respectively (the DMSO content in the reaction solution of the test substance was 5%). Predictor TM hERG Membrane and Predictor TM 1) The hERG Tracer mixture solution was added and reacted for 4 hours. 2) The Gain setting and Z-position (optimal distance between the assay plate and the instrument optics) were set using the Negative control wells, and the G-Factor was calibrated by setting buffer blanks with a value of 50 mP applied to the Free Tracer Control wells. 3) The fluorescence intensity at each concentration (Ex: 530 nm / Em: 590 nm) was measured using Synergy Neo (Biotek, USA).

[0226]

[0227] control group materialClassification Substance Name Manufacturer Solvent control DMS OSigma-Aldrich Positive control E-4031 Invitrogen

[0228] Data analysis was performed as follows: 1) IC was calculated using the mean of the polarization [mP] at each concentration using the fluorescence polarization protocol of Synergy Neo (Biotek USA). 50 Values ​​were derived. 2) IC based on the mean and mean absolute deviation (MAD) by concentration 50 The concentrations were indicated and displayed as a graph. IC of E-4031 (positive control) 50 Since it was analyzed to be 51 nM (reference range: 10 nM~100 nM)3), the standardization of the test system was verified (Fig. 14).

[0229] As a result, the IC of mirodenafil 50 It was analyzed to be 9.91 μM (Fig. 15). This means that mirodenafil inhibits hERG channels in the micromolar concentration range, suggesting that cardiac safety needs to be evaluated by considering the safety margin with respect to plasma concentration during clinical administration.

[0230]

[0231] Experimental Example 2

[0232] Comparison of hERG Safety Margins of Compound 2 and Mirodenafil

[0233] hERG IC, a key factor in predicting cardiotoxicity 50 and maximum plasma concentration (C max This experiment was conducted to determine the safety margin between and to confirm the superiority compared to the control group, mirodenafil.

[0234] hERG IC of Compound 2 and Mirodenafil 50 The value is C measured after oral administration to mice (20 mg / kg). maxThe safety margin was calculated by comparing with ). Ideally, C at human dose max A safety margin should be calculated from ), but at this stage, mouse test results were used to compare with mirodenafil.

[0235] - hERG Safety Margin = hERG IC 50 / Free Plasma C max

[0236] - hERG Safety Margin Ratio = Compound 2 hERG IC 50 / Mirodenafil Free Plasma C max

[0237] IC of E-4031 (positive control) 50 Since it was analyzed to be 26 nM (reference range: 10 nM~100 nM), the standardization of the test system was verified (Fig. 16).

[0238] IC of Compound 2 50 It was analyzed to be 20.22 μM (Fig. 17). From mouse pharmacokinetic results, the hERG safety margin of compound 2 was calculated to be 9.55, which is 3.33 times better than that of mirodenafil (Table 18). This suggests that compound 2 has a significantly lower potential to inhibit hERG channels at the same plasma concentration compared to mirodenafil, and that the risk of cardiotoxicity in clinical practice will be much lower.

[0239] Comparison of hERG Safety Margins of Compound 2 and Mirodenafil Test material hERG IC 50 (μM)C max , Mouse Plasma(20mpk, PO, ng / mL)C max (μM)MWhERG Safety Margin Safety Margin Ratio Compound 220.221,151 2.1254 3.68 9.55 3.33 Mirodenafil 9.91 1.835 3.455 31.68 2.87 1.00

[0240] For reference, sildenafil's hERG IC 50is 33.3 μM, C max It has been reported to be 0.843 μM (Eur J Pharmacol, 2004), and the resulting hERG safety margin is 39.5. No cardiotoxicity issues have been reported in clinical trials for either mirodenafil or sildenafil, suggesting that clinically acceptable cardiac safety can be demonstrated if a safety margin of 2.87 or higher is secured.

[0241] The safety margin (9.55) of Compound 2 is significantly higher than that of Mirodenafil (2.87) and also maintains an appropriate safety margin when compared to Sildenafil (39.5). Therefore, Compound 2 is predicted to be much safer than Mirodenafil in terms of cardiotoxicity in clinical practice.

[0242]

[0243] [Example 6]

[0244] Aβ 1-42 Evaluation of the efficacy of Compound 2 in a mouse model of cognitive impairment induced by [unclear]

[0245] Aβ 1-42 In vivo behavioral pharmacology experiments were performed to verify the memory-enhancing effects of Compound 2 in a mouse model of cognitive impairment induced by Aβ. After administering Compound 2 intraperitoneally once daily for 7 days, its efficacy was evaluated by comparing it to mirodenafil using the Y-maze test. Specifically, as shown in Figure 18, Aβ 1-42 An acute cognitive impairment model was established by injecting into the intracerebroventricular (ICV). Due to the nature of the study, oral administration of the drug could cause excessive stress to mice and affect the test results; therefore, intraperitoneal administration, which causes relatively less stress, was performed. In this model, compound 2 was evaluated against mirodenafil to determine whether it alleviates cognitive impairment and improves memory.

[0246] As shown in Figure 19, the results of the Y-maze experiment revealed a significantly increasing trend in spontaneous alternation in the drug administration group compared to the Aβ1-42(+) vehicle group in One-way ANOVA and Bonferroni multiple comparison tests (*: p < 0.05, ***: p < 0.001). This suggests that both Compound 2 and Mirodenafil improve cognitive function by inhibiting the aggregation of Aβ1-42 monomers injected into the ventricles. In particular, the Compound 2 administration group showed extremely high significance at the p < 0.001 level, clearly demonstrating that it has a statistically superior effect compared to the Mirodenafil administration group, which showed a significance level of p < 0.05. This demonstrates that the compounds of the present invention have the potential to replace and surpass existing substances as treatments for Alzheimer's disease.

[0247] The Arm entry data obtained in this experiment was used as an indicator to confirm that there was no random sampling error caused by differences in baseline activity levels between groups (e.g., individuals with excessively high activity levels being concentrated in a specific group). Since no significant difference in the number of Arm entries was observed between groups, this implies that the experimental system was constructed under a controlled environment with identical activity backgrounds; accordingly, this supports the conclusion that the observed improvement in mouse cognitive function is due to the pure efficacy of the test drug.

Claims

1. Pyrrolopyrimidinone derivatives represented by Chemical Formula 1, pharmaceutically acceptable salts thereof, hydrates thereof, or solvates thereof: [Chemical Formula 1] In the above chemical formula 1, R1 is methyl and; R2 is 4-(oxetan-3-yl).

2. Pyrrolopyrimidinone derivatives represented by Chemical Formula 1, pharmaceutically acceptable salts thereof, hydrates thereof, or solvates thereof: [Chemical Formula 1] In the above chemical formula 1, R1 is ethyl and; R2 is 4-(oxetan-3-yl).

3. A pharmaceutical composition for the prevention or treatment of diseases induced by PDE5 activity, comprising as an active ingredient a pyrrolopyrimidinone derivative of claim 1 or 2, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof.

4. In Paragraph 3, A composition in which the disease induced by the above PDE5 activity is one or more selected from the group consisting of neurological brain disease, cerebrovascular disease, sexual dysfunction, cardiovascular disease, hypertension, and urological disease.

5. In Paragraph 4, A composition wherein the above neurological brain disease is one or more selected from the group consisting of Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), Huntington's disease, frontotemporal dementia, progressive supranuclear palsy, multiple system atrophy, corticobasal degeneration, dementia with Lewy bodies, Charcot-Marie-Tooth disease, and prion diseases.

6. In Paragraph 4, The above-mentioned cerebrovascular disease is a composition comprising one or more selected from the group consisting of ischemic stroke, hemorrhagic stroke, cerebral small vessel disease, cerebral vasospasm, and vascular dementia.

7. In Paragraph 4, The above sexual dysfunction is a composition comprising one or more selected from the group consisting of erectile dysfunction, sexual arousal disorder, premature ejaculation, and decreased sexual satisfaction.

8. In Paragraph 4, The above-mentioned cardiovascular disease is a composition comprising one or more selected from the group consisting of heart failure, cardiac hypertrophy, coronary artery disease, atherosclerosis, peripheral arterial disease, and single ventricular heart, Fontan operation.

9. In Paragraph 4, The above-mentioned hypertension is a composition comprising one or more selected from the group consisting of pulmonary arterial hypertension, essential hypertension, hypertensive heart disease, and renovascular hypertension.

10. In Paragraph 4, The above urological disease is a composition in which benign prostatic hyperplasia is the above.

11. In Paragraph 10, The above-mentioned benign prostatic hyperplasia is Lower Urinary Tract Symptoms (LUTS) A composition of benign prostatic hyperplasia accompanied by 12. In Paragraph 3, The above composition is a composition for oral administration.

13. As shown in Reaction Equation 1, (a) a step of combining a compound represented by chemical formula 2 and a compound represented by chemical formula 3 to form an amide bond, and preparing a compound represented by chemical formula 4 (step 1); (b) a step of reacting the compound represented by Chemical Formula 4 with a base to produce the compound represented by Chemical Formula 5 through a cyclization reaction (Step 2); (c) a step of preparing a compound represented by chemical formula 6 by introducing a sulfonyl chloride group into the compound represented by chemical formula 5 (step 3); and (d) A method for preparing a pyrrolopyrimidinone derivative of claim 1 or 2, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof, comprising the step (step 4) of synthesizing a compound represented by formula 1 by adding a piperazine substituted in a compound represented by formula 6: [Reaction Equation 1] .

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