A class of substituted quinolinone derivatives and their uses

Abstract

This invention discloses a class of quinolinone derivatives and their pharmaceutically acceptable salts. The quinolinone derivatives of this invention are compounds with phosphodiesterase 3A (PDE3A) inhibitory activity, possessing significant potential therapeutic value for use in the treatment of tumors, cardiovascular diseases, cerebrovascular diseases, or dementia.

Description

Technical Field

[0001] This invention relates to the field of medicinal chemistry, and particularly to the use of a class of fluorosubstituted quinolinone derivatives or pharmaceutically acceptable salts thereof as inhibitors of phosphodiesterase 3A (PDE3A), especially in medicaments for the treatment of tumors, cardiovascular diseases, cerebrovascular diseases or dementia. Background Technology

[0002] Cilostazol is a quinolone derivative compound that exerts antiplatelet and vasodilatory effects by inhibiting phosphodiesterase activity in platelets and vascular smooth muscle, thereby increasing cAMP concentration in these tissues. It inhibits ADP, adrenaline, collagen, and arachidonic acid-induced initial and secondary platelet aggregation and release responses, and exhibits significant antithrombotic effects in models of cerebral and peripheral circulatory disturbances induced by collagen, ADP, arachidonic acid, and sodium laurate. It can be used to treat conditions such as atherosclerosis, aortitis, thromboangiitis obliterans, chronic arterial occlusive disease caused by diabetes, cerebrovascular diseases, or dementia.

[0003] The unfavorable pharmacokinetic properties of cilostazol limit its clinical application: In healthy adult men, a single oral dose of 50 mg cilostazol on an empty stomach and after a meal resulted in Cmax and AUCinf that were 2.3 times and 1.4 times higher, respectively, after the meal dose. Cilostazol is primarily metabolized by the cytochrome P450 isoenzyme CYP3A4 in liver microsomes, followed by CYP2D6 and CYP2C19, producing active metabolites such as OPC-13015 (derived through dehydration) and OPC-13213 (derived through hydroxylation). In patients with severe renal impairment, an oral dose of 0.1 g cilostazol daily for 8 consecutive days resulted in a 29% and 39% decrease in Cmax and AUC, respectively, compared to healthy adults, while the Cmax and AUC of the active metabolite OPC-13213 increased by 173% and 209%, respectively. Therefore, it is necessary to design a compound that satisfies both the PDE3A inhibitory activity of cilostazol and has good pharmacokinetic parameters for use in the treatment of related diseases with cilostazol.

[0004] In 1812, French scientists discovered a new element in fluorine (HF) and named it fluorine, marking the first time fluorine (F) was observed in humankind. Over time and with further research, numerous fluorine-containing compounds were discovered and applied, leading to significant advancements in fluorine chemistry. Because the bond energy of the CF bond (487 kJ·mol−1) is higher than that of the CH bond (420 kJ·mol−1), fluorine atoms are the most commonly used blocking groups. In drug design, fluorine atoms are often introduced into small molecule compounds to substitute for fluorine atoms, blocking sites prone to oxidation and selectively preventing oxidative metabolism, thereby improving the metabolic stability of the compound and prolonging the drug's duration of action in vivo. Summary of the Invention

[0005] The purpose of this invention is to provide a quinolinone compound or a pharmaceutically acceptable salt thereof, which has PDE3A inhibitory activity, for the treatment of diseases such as tumors, cardiovascular diseases, cerebrovascular diseases, or dementia.

[0006] To solve the above-mentioned technical problems, the technical solution provided by the present invention is as follows:

[0007] A class of substituted quinolinone derivatives as shown in Formula 1, or their pharmaceutically acceptable salts, hydrates, or solvates:

[0008]

[0009] Formula 1

[0010] Among them, R1, R2, R3, R4, R5, R6, R7, R8, R9 and R10 are each independently hydrogen or halogen; the additional condition is that at least one of R1, R2, R3, R4, R5, R6, R7, R8, R9 or R10 is halogen.

[0011] Preferably, R1, R2, R3, R4, R5, R6, R7, R8, R9 and / or R10 are each independently hydrogen or halogen, wherein at least one of R1, R2, R3, R4, R5, R6, R7, R8, R9 and / or R10 is fluorine.

[0012] More preferably, at least one of R1, R2, R3, R4, R5, R6, R7, R8, R9 and / or R10 is fluorine.

[0013] Most preferably, the compounds of Formula 1 include, but are not limited to, the following specific examples of compounds:

[0014]

[0015] S1

[0016]

[0017] S2

[0018]

[0019] S3

[0020]

[0021] S4

[0022]

[0023] S5

[0024] The compounds provided by this invention also include pharmaceutically acceptable equivalents of the compound or mixtures thereof.

[0025] Preferably, the compounds provided by the present invention may comprise one or a mixture of two or more of pharmaceutically acceptable salts, hydrates, solvates, metabolites, and prodrugs.

[0026] Preferably, the compounds provided by this invention comprise acidic or basic salts of the compounds provided by this invention. The pharmaceutically acceptable salts possess the pharmaceutical activity of the compound and meet the requirements both biologically and practically.

[0027] This invention provides a quinolinone compound or a pharmaceutically acceptable salt thereof for the treatment of diseases such as tumors, cardiovascular diseases, cerebrovascular diseases, or dementia. Detailed Implementation

[0028] This invention discloses quinoline ketone compounds and their uses. Those skilled in the art can refer to the content of this document and appropriately modify the process parameters to achieve the desired results. The methods and applications of this invention have been described through preferred embodiments. It is evident that those skilled in the art can modify or appropriately change and combine the methods and applications described herein without departing from the content, spirit, and scope of this invention to realize and apply the technology of this invention.

[0029] To enable those skilled in the art to better understand the technical solutions of the present invention, the present invention will be further described in detail below with reference to specific embodiments.

[0030] The compounds of the general formula of this invention can be synthesized via the following route:

[0031]

[0032] Example 1: Synthesis of compound S1

[0033]

[0034]

[0035] first step:

[0036] 0.5 g (1 eq 2.92 mmol) of m-difluorocyclohexylamine hydrochloride was dissolved in DCM, and 0.74 g (2.5 eq 7.3 mmol) of TEA was added. The mixture was cooled to 0°C on an ice bath, and 0.67 g (1.5 eq 4.38 mmol) of acyl chloride was added dropwise. The reaction was carried out overnight at room temperature. After the reaction was completed, the mixture was washed three times each with 250 mM HCl and saturated NaHCO3. The organic phase was dried and concentrated (30°C) to give 0.74 g of white solid, with a yield of 100% (theoretical).

[0037] Step Two;

[0038] In the first step, the product was dissolved in Tol, and the mixture was kept in an ice bath at 0°C. PCl5 0.85 g (1.4 eq 4.1 mmol) was added in portions, and the mixture was stirred at room temperature for 2 h after the addition was complete. Then, TMSN3 1.17 g (3.5 eq 10.22 mmol) was added at room temperature, and the mixture was stirred at room temperature for 30 min. The temperature was then raised to 80°C and reacted for 3 h. After the reaction was complete, EA was added for dilution, and the organic phase was washed three times with saturated NaHCO3. The organic phase was then dried and concentrated (30°C). The yield was 83% (0.67 g).

[0039] Step 3;

[0040] Take 0.58 g (1.5 eq 3.63 mmol) of 7-hydroxyquinoline, add 0.19 g (1.4 eq 3.39 mmol) of KOH, add 10 ml of DMF, heat to 90 degrees, add 0.67 g (1 eq 2.42 mmol) of the product from step 2, then add 80 mg of KI (0.2 eq 0.48 mmol), react at 90 degrees for 8 h, dilute the organic phase with EA, and wash twice with 0.5 N NaOH. Dry and concentrate the organic phase (30 degrees). Purify the crude product with 200-300 mesh silica gel (HE:EA = 4:1, start gradient elution until HE:EA = 1:4, collect the product, concentrate, and before completely dry, add HE to crystallize, filter to obtain a white solid) 0.35 g 30% yield.

[0041] ESI-MS: 404.2 [M+H]+

[0042] 1H NMR (400 MHz, CDCl3) δ11.2-11.3 (s, 1H), 7.7 (d, J=9.4Hz, 1H), 7.5 (d, J=8.7Hz, 1H), 6.7 (m, 2H), 6.55 (d, J=9.4Hz, 1H), 4.4 (dq, J=12.9, 7.7,5.4Hz, 1H), 4.1 (t, J=5.9Hz, 2H), 2.9 (t, J=7.5Hz, 2H), 2.6-2.4 (m, 2H), 2.3-2.1 (m, 2H), 2.1-1.9(m, 8H).

[0043] Example 2: Synthesis of compound S2

[0044]

[0045]

[0046] first step:

[0047] 0.446 g (1 eq 2.92 mmol) of m-fluorocyclohexylamine hydrochloride was dissolved in 50 mL of DCM, and 0.74 g (2.5 eq 7.3 mmol) of TEA was added. The mixture was cooled to 0°C on an ice bath, and 0.67 g (1.5 eq 4.38 mmol) of acyl chloride was added dropwise. The reaction was allowed to proceed overnight at room temperature. After the reaction was complete, the mixture was washed three times each with 250 mM HCl and saturated NaHCO3. The organic phase was dried and concentrated (30°C) to give 0.686 g of a white solid, with a yield of 100% (theoretical).

[0048] Step Two;

[0049] In the first step, the product was dissolved in 50 mL of Tol, and the mixture was kept in an ice bath at 0°C. PCl5 0.85 g (1.4 eq 4.1 mmol) was added in portions, and the mixture was stirred at room temperature for 2 h after the addition was complete. Then, TMSN3 1.17 g (3.5 eq 10.22 mmol) was added at room temperature, and the mixture was stirred at room temperature for 30 min. The temperature was then raised to 80°C and reacted for 3 h. After the reaction was complete, EA was added for dilution, and the organic phase was washed three times with saturated NaHCO3. The organic phase was then dried and concentrated (30°C). The yield was 83% (0.63 g).

[0050] Step 3;

[0051] Take 0.58 g (1.5 eq 3.63 mmol) of quinolinone, add 0.19 g (1.4 eq 3.39 mmol) of KOH, add 10 ml of DMF, heat to 90 degrees Celsius until dissolved, add 0.63 g (1 eq) of the product from step 2 and 80 mg (0.2 eq 0.48 mmol) of KI, react at 90 degrees Celsius for 8 h, dilute the organic phase with EA, and wash twice with 0.5 N NaOH, dry and concentrate the organic phase (30 degrees Celsius), purify the crude product with 200-300 mesh silica gel (HE:EA = 4:1, start gradient elution, until HE:EA = 1:4, collect the product, concentrate, and when not completely dry, add HE to precipitate crystals, filter to obtain a white solid) 0.34 g 30% yield.

[0052] ESI-MS: 359.2 [M+H]+

[0053] 1H NMR (400 MHz, CDCl3) δ11.2-11.3 (s, 1H), 7.7(d, J=9.4Hz, 1H), 7.5(d, J=8.7Hz, 1H), 6.7(m, 2H), 6.55 (d, J=9.4Hz, 1H), 4.55(m, 1H), 4.2 (t, J=5.9Hz, 2H), 3.0 (t, J=7.5Hz, 2H), 2.3-2.0 (m, 8H), 1.9 (m. 2H), 1.8 (m, 2H).

[0054] Example 3: Synthesis of compound S3

[0055]

[0056]

[0057] first step:

[0058] 0.5 g (1 eq 2.92 mmol) of p-difluorocyclohexylamine hydrochloride was dissolved in 50 ml of DCM, and 0.74 g (2.5 eq 7.3 mmol) of TEA was added. The mixture was cooled to 0°C on an ice bath, and 0.67 g (1.5 eq 4.38 mmol) of acyl chloride was added dropwise. The reaction was allowed to proceed overnight at room temperature. After the reaction was complete, the mixture was washed three times each with 250 mM HCl and saturated NaHCO3. The organic phase was dried and concentrated (30°C) to give a white solid, 0.74 g, with a 100% yield (theoretical).

[0059] Step Two;

[0060] In the first step, the product was dissolved in 50 ml of Tol, and the mixture was kept in an ice bath at 0°C. 0.85 g of PCl5 (1.4 eq 4.1 mmol) was added in portions, and the mixture was stirred at room temperature for 2 hours after the addition was complete. Then, 1.17 g of TMSN3 (3.5 eq 10.22 mmol) was added at room temperature, and the mixture was stirred at room temperature for 30 minutes. The temperature was then raised to 80°C and reacted for 3 hours. After the reaction was complete, EA was added for dilution, and the organic phase was washed three times with saturated NaHCO3. The organic phase was then dried and concentrated (at 30°C). The yield was 83% (0.67 g).

[0061] Step 3;

[0062] Take 0.58 g of quinolinone (1.5 eq 3.63 mmol), add 0.19 g of KOH (1.4 eq 3.39 mmol), add 10 ml of DMF, heat to 90 degrees Celsius until dissolved, add 0.67 g of the product from step 2 (1 eq 2.42 mmol) and 80 mg of KI (0.2 eq 0.48 mmol), react at 90 degrees Celsius for 8 h, dilute the organic phase with EA, and wash twice with 0.5 N NaOH, dry and concentrate the organic phase (30 degrees Celsius), purify the crude product with 200-300 mesh silica gel (HE:EA = 4:1, start gradient elution, until HE:EA = 1:4, collect the product, concentrate, and before completely dry, add HE to precipitate crystals, filter to obtain a white solid) 0.35 g 30% yield.

[0063] ESI-MS: 404.2 [M+H]+

[0064] 1H NMR (400 MHz, CDCl3) δ11.2-11.3 (s, 1H), 7.7 (dd, J=9.4, 2.2Hz,1H), 7.4 (dd, J=8.8, 2.2Hz, 1H), 6.75-6.65 (m, 2H), 6.45 (dd, J=9.4, 2.3Hz,1H), 4.35 (m. 1H), 4.1 (td, J=5.9, 2.1Hz,2H), 2.9 (td, J=7.5, 2.2Hz, 2H), 2.4-1.7 (m, 12H).

[0065] Example 4: Synthesis of compound S4

[0066]

[0067]

[0068] first step:

[0069] 0.45 g (1 eq 2.92 mmol) of o-fluorocyclohexylamine hydrochloride was dissolved in 50 ml of DCM, and 0.74 g of TEA (2.5 eq 7.3 mmol) was added. The mixture was cooled to 0°C on an ice bath, and 0.67 g (1.5 eq 4.38 mmol) of acyl chloride was added dropwise. The reaction was allowed to proceed overnight at room temperature. After the reaction was complete, the mixture was washed three times each with 250 mM HCl and saturated NaHCO3. The organic phase was dried and concentrated (30°C) to give 0.69 g of a white solid, with a yield of 100% (theoretical).

[0070] Step Two;

[0071] The first step involved dissolving the product in 50 ml of Tol, cooling to 0°C on ice, and adding 0.85 g of PCl5 (1.4 eq 4.1 mmol) in portions. After the addition was complete, the mixture was stirred at room temperature for 2 hours. Then, 1.17 g of TMSN3 (3.5 eq 10.22 mmol) was added at room temperature, and the mixture was stirred at room temperature for 30 minutes. The temperature was then raised to 80°C and reacted for 3 hours. After the reaction was complete, EA was added for dilution, and the organic phase was washed three times with saturated NaHCO3. The organic phase was then dried and concentrated (at 30°C). The yield was 83% (0.63 g).

[0072] Step 3;

[0073] Take 0.58 g of quinolinone (1.5 eq 3.63 mmol), add 0.19 g of KOH (1.4 eq 3.39 mmol), add 10 ml of DMF, heat to 90 degrees Celsius until dissolved, add 0.63 g (1 eq) of the product from step 2 and 80 mg of KI (0.2 eq 0.48 mmol), react at 90 degrees Celsius for 8 h, dilute the organic phase with EA, and wash twice with 0.5 N NaOH, dry and concentrate the organic phase (30 degrees Celsius), purify the crude product with 200-300 mesh silica gel (HE:EA = 4:1, start gradient elution, until HE:EA = 1:4, collect the product, concentrate, and when not completely dry, add HE to precipitate crystals, filter to obtain a white solid) 0.34 g 30% yield.

[0074] ESI-MS: 359.2 [M+H]+

[0075] 1H NMR (400 MHz, MeOH-D4) δ7.8 (d, J=9.3Hz, 1H), 7.6 (d, J=8.7Hz,1H), 6.9 (m, 2H), 6.4 (d, J=9.4Hz, 1H), 4.5 (m, 2H), 4.2 (t, J=6.0Hz, 2H),3.1 (t, J=7.5Hz, 2H), 2.4 (m, 1H), 2.2-2.1 (m, 2H), 2.1-2.0 (m, 2H), 2.0-1.8(m. 4H), 1.75-1.6 (m, 1H), 1.6-1.45 (m, 2H).

[0076] Example 5: Synthesis of compound S5

[0077]

[0078]

[0079] first step:

[0080] 0.45 g (1 eq 2.92 mmol) of o-fluorocyclohexylamine hydrochloride was dissolved in 50 ml of DCM, and 0.74 g of TEA (2.5 eq 7.3 mmol) was added. The mixture was cooled to 0°C on an ice bath, and 0.67 g (1.5 eq 4.38 mmol) of acyl chloride was added dropwise. The reaction was allowed to proceed overnight at room temperature. After the reaction was complete, the mixture was washed three times each with 250 mM HCl and saturated NaHCO3. The organic phase was dried and concentrated (30°C) to give 0.69 g of a white solid, with a yield of 100% (theoretical).

[0081] Step Two;

[0082] The first step involved dissolving the product in 50 ml of Tol, cooling to 0°C on ice, and adding 0.85 g of PCl5 (1.4 eq 4.1 mmol) in portions. After the addition was complete, the mixture was stirred at room temperature for 2 hours. Then, 1.17 g of TMSN3 (3.5 eq 10.22 mmol) was added at room temperature, and the mixture was stirred at room temperature for 30 minutes. The temperature was then raised to 80°C and reacted for 3 hours. After the reaction was complete, EA was added for dilution, and the organic phase was washed three times with saturated NaHCO3. The organic phase was then dried and concentrated (at 30°C). The yield was 83% (0.63 g).

[0083] Step 3;

[0084] Take 0.58 g of quinolinone (1.5 eq 3.63 mmol), add 0.19 g of KOH (1.4 eq 3.39 mmol), add 10 ml of DMF, heat to 90 degrees Celsius until dissolved, add 0.63 g (1 eq) of the product from step 2 and 80 mg of KI (0.2 eq 0.48 mmol), react at 90 degrees Celsius for 8 h, dilute the organic phase with EA, and wash twice with 0.5 N NaOH, dry and concentrate the organic phase (30 degrees Celsius), purify the crude product with 200-300 mesh silica gel (HE:EA = 4:1, start gradient elution, until HE:EA = 1:4, collect the product, concentrate, and when not completely dry, add HE to precipitate crystals, filter to obtain a white solid) 0.34 g 30% yield.

[0085] ESI-MS: 359.2 [M+H]+

[0086] 1H NMR (400 MHz, MeOH-D4) δ 7.9 (d, J=9.4Hz, 1H), 7.6 (d, J=9.4Hz,1H), 6.9 (m, 2Hz), 6.4 (d, J=9.4Hz, 1H), 4.7 -4.5 (m. 2H), 4.2 (t, J=5.9Hz,2H), 3.1 (t, J=7.5Hz, 2H), 2.3 (m, 1H), 2.2-2.1 (m, 2H), 2.1-2.0 (m, 2H), 2.0-1.8 (m, 4H), 1.75-1.6 (m, 1H), 1.6-1.45 (m, 2H).

[0087] Example 6: Detection of the inhibitory activity of the compound against phosphodiesterase 3A (PDE3A)

[0088] PDE3A activity was detected using the PDE3A TR-FRET Assay Kit (BPS Catalog # 60706). This kit is designed to identify PDE3A inhibitors using TR-FRET (time-resolved fluorescence resonance energy transfer) technology. The assay is based on FAM-labeled nucleotide monophosphates produced by phosphodiesterase. These phosphate groups bind to Tb-labeled nanoparticles, causing energy transfer from Tb to FAM, which emits a fluorescent signal at 520 nm. Changes in fluorescence intensity can be easily measured using a multi-mode microplate reader.

[0089] Experimental steps:

[0090] 1) Dilute 20 μM MFAAM-Cyclic-3',5'-AMP substrate stock solution 100-fold with PDE buffer to prepare a 200 nM solution. Perform only a sufficient amount of analysis; aliquot the remaining stock solution and store at -20°C.

[0091] 2) Add 25 μl of FAM-Cyclic-3',5'-AMP (200 nM) to each well labeled “Substrate Control,” “Positive Control,” and “Test Inhibitor.” Add 25 μl of PDE analysis buffer to each well designated “Tb Control Only.”

[0092] 3) Add 5 μl of inhibitor solution to each well designated as "Test Inhibitor". Add 5 μl of the same solution without inhibitor (inhibitor buffer) to "Tb Control Only", "Substrate Control", and "Positive Control".

[0093] 4) Thaw PDE3A on ice. After the first thaw, briefly rotate the test tube containing the enzyme to restore its full contents. Aliquot the PDE3A enzyme for single use. Immediately store the remaining undiluted enzyme in equal portions at -80°C.

[0094] 5) Dilute PDE3A to 0.05 ng / μl (1 ng / reaction) in PDE buffer. Add 20 μl of PDE analysis buffer to the wells designated "Tb Control Only" and "Substrate Control," and add 20 μl of PDE3A (0.05 ng / μl) to the well designated "Positive Control" to start the reaction. Discard any remaining diluted enzyme after use.

[0095] 6) Incubate at room temperature for 1 hour.

[0096] 7) Prepare a binding dilution buffer by mixing equal volumes of binding buffer A and binding buffer B.

[0097] 8) Mix the adhesive thoroughly and dilute it with the binding dilution buffer prepared in step 1 at a ratio of 1:50.

[0098] 9) Add Tb donor to the mixture (1:1000 dilution).

[0099] 10) Add 100 μl to each well. Incubate at room temperature with gentle shaking for 1 hour.

[0100] 11) Read the fluorescence intensity in a microtiter plate reader with TR-FRET function.

[0101] Calculation results:

[0102] (1) Fluorescence intensity calculation

[0103]

[0104] Where S520 = sample 520 nm reading, S490 = sample 490 nm reading, Tb520 = Tb 520 nm reading only, and Tb490 = Tb 490 nm reading only. When calculating the percentage of activity, the FRET value for the substrate control only can be set to zero activity, and the FRET value for the positive control can be set to 100% activity.

[0105] (2) Calculation of enzyme activity inhibition rate

[0106] × 100

[0107] Where FRETs = sample FRET, FRETSub = substrate control FRET only, and FRETP = positive control FRET.

[0108] (3) Calculation of IC50 value:

[0109] Using log[compound concentration] as the x-axis and Inhibition % as the y-axis, fit a nonlinear curve in GraphPad Prism 7: log (inhibitor) vs. response -- Variable slope, and calculate the IC50 value.

[0110] The results are shown in the table. All synthesized compounds S1-S5 have PDE3A enzyme inhibitory activity.

[0111] S1 S2 S3 S4 S5 IC50 value ++ ++ +++ +++ +++

[0112] Note: ++ indicates an IC50 value between 1 μM and 5 μM; ++ indicates an IC50 value between 5 μM and 10 μM; + indicates an IC50 value greater than 10 μM.

Claims

1. A quinolinone compound or a pharmaceutically acceptable salt thereof, characterized by, The compound is a compound selected from the group consisting of: S3 S4 S5。 2. Use of the compound or pharmaceutically acceptable salt thereof according to claim 1 in the preparation of an antitumor, cardiovascular disease, cerebrovascular disease or dementia treating drug.

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