Substituted quinolinone derivative and application thereof

Abstract

The invention discloses a quinolinone derivative and a pharmaceutically acceptable salt of the quinolinone derivative. The quinolinone derivative disclosed by the invention is a compound with phosphodiesterase 3A (PDE3A) inhibition activity, has an important potential treatment value, and is applied to medicines for treating tumors, cardiovascular diseases, cerebrovascular diseases or dementia.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical chemistry, and in particular to a class of fluorine-substituted quinolinone derivatives or pharmaceutically acceptable salts thereof, and their use as phosphodiesterase 3A (PDE3A) inhibitors, in particular in drugs for treating tumors, cardiovascular diseases, cerebrovascular diseases or dementia. Background Art

[0002] Cilostazol is a quinolinone derivative compound that can inhibit the activity of phosphodiesterase in platelets and vascular smooth muscle, thereby increasing the cAMP concentration in platelets and smooth muscle, exerting antiplatelet and vasodilatory effects. It inhibits the initial and secondary aggregation and release reactions of platelets induced by ADP, adrenaline, collagen and arachidonic acid, and has a significant antithrombotic effect on the model of cerebral circulation and peripheral circulation disorders caused by collagen, ADP, arachidonic acid and sodium laurate. It can be used to treat chronic arterial occlusive disease caused by atherosclerosis, large arteritis, thromboangiitis obliterans, diabetes, cerebrovascular disease or dementia.

[0003] The poor pharmacokinetic properties of cilostazol limit its clinical application: when a single oral dose of cilostazol 50 mg was given to healthy adult men on an empty stomach and after a meal, the Cmax and AUCinf of the post-meal administration were 2.3 times and 1.4 times those of the fasting state, respectively. Cilostazol is mainly metabolized by the cytochrome P450 isoenzyme CYP3A4 in liver microsomes, followed by CYP2D6 and CYP2C19, and metabolized into active metabolites such as OPC-13015 generated by dehydration and OPC-13213 generated by hydroxylation. When patients with severe renal dysfunction took cilostazol 0.1 g per day orally for 8 consecutive days, the Cmax and AUC of cilostazol decreased by 29% and 39%, respectively, compared with 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 not only satisfies the PDE3A inhibitory activity of cilostazol but also has good pharmacokinetic parameters for use in diseases related to cilostazol treatment.

[0004] In 1812, French scientists discovered a new element in HF and named it Fluorine. This was the first time that the F element appeared in front of humans. With the passage of time and the deepening of research, a large number of fluorine-containing compounds have been discovered and applied, and fluorine chemistry has also made great progress. Since the bond energy of the CF bond (487 kJ·mol−1) is higher than that of the CH bond (420 kJ·mol−1), the fluorine atom is the most commonly used blocking group. In drug design, fluorine atoms are usually introduced into small molecule compounds to block sites that are prone to oxidative metabolism, selectively prevent the occurrence of oxidative metabolism, and thus improve the metabolic stability of the compound and prolong the drug's action time in the body. Summary of the invention

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

[0006] In order to solve the above technical problems, the technical solution provided by the present invention is:

[0007] A substituted quinolone derivative as shown in Formula 1 or a pharmaceutically acceptable salt, hydrate or solvate thereof:

[0008]

[0009] Formula 1

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

[0011] Preferably, it is characterized in that 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, it is characterized in that at least one of R1, R2, R3, R4, R5, R6, R7, R8, R9 and / or R10 is fluorine.

[0013] Most preferably, the compound of formula 1 includes but is not limited to the following specific compound examples:

[0014]

[0015] S1

[0016]

[0017] S2

[0018]

[0019] S3

[0020]

[0021] S4

[0022]

[0023] S5

[0024] The compounds provided by the present invention also include pharmaceutically acceptable equivalents of the compounds or a mixture of two or more.

[0025] Preferably, the compound provided by the present invention may include one or a mixture of two or more of a pharmaceutically acceptable salt, a hydrate, a solvate, a metabolite, or a prodrug.

[0026] Preferably, the compound provided by the present invention comprises an acid salt or a basic salt of the compound provided by the present invention. The pharmaceutically acceptable salt has the pharmaceutical activity of the compound and meets the requirements in biology and practical applications.

[0027] The present invention provides a quinolinone compound or a pharmaceutically acceptable salt thereof for treating diseases such as tumors, cardiovascular diseases, cerebrovascular diseases or dementia. DETAILED DESCRIPTION

[0028] The present invention discloses quinolinone compounds and their uses. Those skilled in the art can refer to the content of this article and appropriately improve the process parameters to achieve the desired effect. The method and application of the present invention have been described through preferred embodiments. It is obvious that relevant personnel can modify or appropriately change and combine the method and application described herein without departing from the content, spirit and scope of the present invention to achieve and apply the technology of the present invention.

[0029] In order to enable those skilled in the art to better understand the technical solution of the present invention, the present invention is further described in detail below in conjunction with specific embodiments.

[0030] The general formula compound of the present 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 in an ice bath, and 0.67 g (1.5 eq 4.38 mmol) of acid chloride was added dropwise. The mixture was reacted at room temperature overnight. After reaction, the mixture was washed with 250 mM HCl and saturated NaHCO3 for 3 times each. The organic phase was dried and concentrated (30°C) to obtain 0.74 g of a white solid with a yield of 100% (theoretical).

[0037] Step 2:

[0038] The first step product was dissolved in Tol, ice-bathed to 0 degrees, and PCl5 0.85g (1.4eq 4.1mmol) was added in batches. After the addition, it was stirred at room temperature for 2h, and then TMSN3 1.17g (3.5eq 10.22mmol) was added at room temperature. It was stirred at room temperature for 30min, and the temperature was raised to 80 degrees to react for 3h. After the end, EA was added to dilute, and the organic phase was washed with saturated NaHCO3 for 3 times, and the organic phase was dried and concentrated (30 degrees). 0.67g yield 83%.

[0039] Step 3;

[0040] Take 0.58g (1.5eq 3.63mmol) of 7-hydroxyquinolinone, add 0.19g (1.4eq 3.39mmol) of KOH, add 10ml of DMF, heat to 90 degrees, add 0.67g (1eq 2.42mmol) of the second step product, then add 80mg KI (0.2eq0.48mmol), react at 90 degrees for 8h, add EA to dilute the organic phase, and wash it twice with 0.5N NaOH. The organic phase is dried and concentrated (30 degrees), and the crude product is purified with 200~300 mesh silica gel (HE:EA=4:1, start gradient elution to HE:EA=1:4, collect the product, concentrate, add HE for crystallization when it is not completely concentrated, and filter to obtain a white solid) 0.35g 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 in an ice bath, and 0.67 g (1.5 eq 4.38 mmol) of acid chloride was added dropwise. The mixture was reacted at room temperature overnight. After reaction, the mixture was washed with 250 mM HCl and saturated NaHCO3 for 3 times each. The organic phase was dried and concentrated (30°C) to obtain 0.686 g of a white solid with a yield of 100% (theoretical).

[0048] Step 2:

[0049] The first step product was dissolved in Tol 50ML, ice-bathed to 0 degrees, and PCl5 0.85g (1.4eq 4.1mmol) was added in batches. After the addition, it was stirred at room temperature for 2 hours, and then TMSN3 1.17g (3.5eq 10.22mmol) was added at room temperature, stirred at room temperature for 30 minutes, and the temperature was raised to 80 degrees for 3 hours. After the end, EA was added to dilute, and the organic phase was washed with saturated NaHCO3 for 3 times, and the organic phase was dried and concentrated (30 degrees). 0.63g yield 83%.

[0050] Step 3;

[0051] Take 0.58g (1.5eq 3.63mmol) of quinolinone, add 0.19g (1.4eq 3.39mmol) of KOH, add 10ml of DMF, heat to 90 degrees until dissolved, add 0.63g (1eq) of the second step product and 80mg (0.2eq 0.48mmol) of KI, react at 90 degrees for 8h, add EA to dilute the organic phase, and wash it twice with 0.5N NaOH. The organic phase is dried and concentrated (30 degrees), and the crude product is purified with 200-300 mesh silica gel (HE:EA=4:1, start gradient elution to HE:EA=1:4, collect the product, concentrate, add HE to crystallize when it is not completely concentrated, and filter to obtain a white solid) 0.34g 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] Dissolve 0.5 g (1 eq 2.92 mmol) of p-difluorocyclohexylamine hydrochloride in 50 ml of DCM, add 0.74 g (2.5 eq 7.3 mmol) of TEA, cool to 0 degrees in an ice bath, add 0.67 g (1.5 eq 4.38 mmol) of acid chloride dropwise, and react at room temperature overnight. After reaction, wash with 250 mM HCl and saturated NaHCO3 three times each. Dry and concentrate the organic phase (30 degrees) to obtain a white solid, 0.74 g, yield 100% (theoretical).

[0059] Step 2:

[0060] The first step product was dissolved in 50ml of Tol, ice-bathed to 0 degrees, and 0.85g PCl5 (1.4eq 4.1mmol) was added in batches. After the addition, it was stirred at room temperature for 2 hours, and then 1.17g TMSN3 (3.5eq 10.22mmol) was added at room temperature, stirred at room temperature for 30 minutes, and the temperature was raised to 80 degrees for 3 hours. After the end, EA was added to dilute, and the organic phase was washed with saturated NaHCO3 for 3 times, and the organic phase was dried and concentrated (30 degrees). 0.67g yield 83%.

[0061] Step 3;

[0062] Take 0.58g (1.5eq 3.63mmol) of quinolinone, add 0.19g KOH (1.4eq 3.39mmol), add 10ml DMF, heat to 90 degrees until dissolved, add 0.67g (1eq, 2.42mmol) of the second step product and 80mg KI (0.2eq0.48mmol), react at 90 degrees for 8h, add EA to dilute the organic phase, and wash it twice with 0.5N NaOH. The organic phase is dried and concentrated (30 degrees), and the crude product is purified with 200-300 mesh silica gel (HE:EA=4:1, start gradient elution to HE:EA=1:4, collect the product, concentrate, add HE to crystallize when it is not completely concentrated, and filter to obtain a white solid) 0.35g 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-monofluorocyclohexylamine 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 in an ice bath, and 0.67 g (1.5 eq 4.38 mmol) of acid chloride was added dropwise. The mixture was reacted at room temperature overnight. After reaction, the mixture was washed with 250 mM HCl and saturated NaHCO3 for 3 times each. The organic phase was dried and concentrated (30°C) to obtain 0.69 g of a white solid with a yield of 100% (theoretical).

[0070] Step 2:

[0071] The first step product was dissolved in 50ml of Tol, ice-bathed to 0 degrees, and 0.85g PCl5 (1.4eq 4.1mmol) was added in batches. After the addition, it was stirred at room temperature for 2h, and then 1.17g TMSN3 (3.5eq 10.22mmol) was added at room temperature. It was stirred at room temperature for 30min, and the temperature was raised to 80 degrees to react for 3h. After the end, EA was added to dilute, and the organic phase was washed with saturated NaHCO3 for 3 times, and the organic phase was dried and concentrated (30 degrees). 0.63g, yield 83%.

[0072] Step 3;

[0073] Take 0.58g (1.5eq 3.63mmol) of quinolinone, add 0.19g KOH (1.4eq 3.39mmol), add 10ml DMF, heat to 90 degrees until dissolved, add 0.63g (1eq) of the second step product and 80mg KI (0.2eq 0.48mmol), react at 90 degrees for 8h, add EA to dilute the organic phase, and wash it twice with 0.5N NaOH, dry and concentrate the organic phase (30 degrees), and purify the crude product with 200-300 mesh silica gel (HE:EA=4:1, start gradient elution to HE:EA=1:4, collect the product, concentrate, add HE to crystallize when it is not completely concentrated, and filter to obtain a white solid) 0.34g 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-monofluorocyclohexylamine 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 in an ice bath, and 0.67 g (1.5 eq 4.38 mmol) of acid chloride was added dropwise. The mixture was reacted at room temperature overnight. After reaction, the mixture was washed with 250 mM HCl and saturated NaHCO3 for 3 times each. The organic phase was dried and concentrated (30°C) to obtain 0.69 g of a white solid with a yield of 100% (theoretical).

[0081] Step 2:

[0082] The first step product was dissolved in 50ml of Tol, ice-bathed to 0 degrees, and 0.85g PCl5 (1.4eq 4.1mmol) was added in batches. After the addition, it was stirred at room temperature for 2h, and then 1.17g TMSN3 (3.5eq 10.22mmol) was added at room temperature. It was stirred at room temperature for 30min, and the temperature was raised to 80 degrees to react for 3h. After the end, EA was added to dilute, and the organic phase was washed with saturated NaHCO3 for 3 times, and the organic phase was dried and concentrated (30 degrees). 0.63g, yield 83%.

[0083] Step 3;

[0084] Take 0.58g (1.5eq 3.63mmol) of quinolinone, add 0.19g KOH (1.4eq 3.39mmol), add 10ml DMF, heat to 90 degrees until dissolved, add 0.63g (1eq) of the second step product and 80mg KI (0.2eq 0.48mmol), react at 90 degrees for 8h, add EA to dilute the organic phase, and wash it twice with 0.5N NaOH, dry and concentrate the organic phase (30 degrees), and purify the crude product with 200-300 mesh silica gel (HE:EA=4:1, start gradient elution to HE:EA=1:4, collect the product, concentrate, add HE to crystallize when it is not completely concentrated, and filter to obtain a white solid) 0.34g 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 compounds on phosphodiesterase 3A (PDE3A)

[0088] PDE3A activity was detected using the PDE3A TR-FRET Assay Kit (BPS Catalog # 60706), which 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 generated by phosphodiesterases. These phosphate groups bind to Tb-labeled nanoparticles, resulting in energy transfer from Tb to FAM, which emits a fluorescent signal at 520 nm. The change in fluorescence intensity can be easily measured using a multi-function microplate reader.

[0089] Experimental steps:

[0090] 1) Dilute 20 μM FAM-Cyclic-3',5'-AMP substrate stock solution 100-fold with PDE buffer to make a 200 nM solution. Use only enough to perform the assay; store the remaining stock solution in aliquots 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 Assay Buffer to each well designated as "Tb Only Control."

[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 the "Tb Only Control", "Substrate Control", and "Positive Control".

[0093] 4) Thaw PDE3A on ice. After the first thaw, briefly spin the tube containing the enzyme to recover the entire contents of the tube. Aliquot the PDE3A enzyme into single-use aliquots. Immediately aliquot remaining undiluted enzyme and store at -80°C.

[0094] 5) Dilute PDE3A to 0.05 ng / μl (1 ng / reaction) in PDE buffer. Start the reaction by adding 20 μl of PDE assay buffer to the wells designated as "Tb only control" and "substrate control", and 20 μl of PDE3A (0.05 ng / μl) to the wells designated as "positive control" and "test inhibitor", discarding any remaining diluted enzyme after use.

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

[0096] 7) Make Binding Dilution Buffer by mixing equal volumes of Binding Buffer A and Binding Buffer B.

[0097] 8) Mix the binder thoroughly and dilute the binder 1:50 with the binding dilution buffer prepared in step 1.

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

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

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

[0101] Calculation results:

[0102] (1) Calculation of fluorescence intensity

[0103]

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

[0108] (3) Calculation of IC50 value:

[0109] With log[compound concentration] as the horizontal axis and Inhibition % as the vertical axis, fit the 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, and the synthesized compounds S1-S5 all have PDE3A enzyme inhibitory activity.

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

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

Claims

1. A quinolinone compound as shown in Formula 1 or a pharmaceutically acceptable salt, hydrate or solvate thereof: Formula 1 in, R1, R2, R3, R4, R5, R6, R7, R8, R9 and R10 are each independently hydrogen or halogen; with the additional proviso that at least one of R1, R2, R3, R4, R5, R6, R7, R8, R9 or R10 is halogen.

2. Any compound according to claim 1, characterized in that 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.

3. Any compound according to claim 1, characterized in that At least one of R1, R2, R3, R4, R5, R6, R7, R8, R9 and / or R10 is fluorine.

4. The compound according to claim 1, characterized in that The compound is a compound selected from the following group or a pharmaceutically acceptable salt thereof: S1 S2 S3 S4 S5。 5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The compounds described do not include non-fluorinated compounds.

6. The compound according to claims 1 to 4 or a pharmaceutically acceptable salt thereof, characterized in that: The compounds have phosphodiesterase 3A (PDE3A) inhibitory activity for use in medicines.

7. Use of the compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof in the preparation of an anti-tumor drug, a drug for treating cardiovascular disease, cerebrovascular disease or dementia.

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