Paldostanol derivative as well as preparation method and application thereof

Abstract

The invention relates to a pardoprunox derivative, a preparation method of the pardoprunox derivative, and an application of the pardoprunox derivative. Specifically disclosed is a compound represented by formula I or a pharmaceutically acceptable salt thereof. The compound provided by the invention has good tumor inhibition activity.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a pardorunox derivative, its preparation method, and its application. Background Technology

[0002] Pardolunol was an innovative drug that was once in the research and development stage and was initially designed to treat Parkinson's disease. It represents a unique mechanism of action in the field of neuropharmacology: a "partial dopamine receptor agonist" and a "5-HT2O" agent. 1A "The dual mode of action of receptor full agonists."

[0003]

[0004] In the treatment of Parkinson's disease, the core pathology is the degeneration of dopaminergic neurons in the brain, leading to dopamine deficiency and triggering motor symptoms (such as tremor, rigidity, and bradykinesia). While traditional levodopa preparations are effective, long-term use can cause serious motor complications such as the "wearing-off phenomenon" and "dyskinesia." Paldolunocal, as a partial agonist of dopamine D2 / D3 receptors, can moderately activate the dopamine pathway to improve symptoms, and its relatively weak and stable activation effect theoretically reduces the risk of dyskinesia. Simultaneously, it stimulates 5-HT... 1A The receptor is believed to help alleviate the depression and anxiety symptoms often associated with Parkinson's disease and may also have a regulatory effect on certain motor complications.

[0005] There are few reports of antitumor activity in petorunol. Summary of the Invention

[0006] Objectives of the invention: The first objective of this invention is to provide a class of paldoluno derivatives; the second objective of this invention is to provide a method for preparing the paldoluno derivatives; and the third objective of this invention is to provide applications of the paldoluno derivatives.

[0007] Technical solution: The paldoluno derivative of this invention has the following structural formula:

[0008]

[0009] Where n is 1 to 5.

[0010] Preferably, the structural formula of the paldoluno derivative is:

[0011]

[0012] The method for synthesizing paldolino derivative (I) according to the present invention includes the following steps:

[0013] (1) Compound 2 and pomalidomide undergo a nucleophilic substitution reaction under the catalysis of a base reagent to give compound 3;

[0014] (2) Compound 3 was hydrolyzed from Boc by a strong acid reagent to obtain compound 4;

[0015] (2) Compound 4 and compound 5 were amidated to give final product I.

[0016]

[0017] Where n is 1 to 5.

[0018] The solvent used in step (1) is an organic solvent, such as tetrahydrofuran, N,N-dimethylformamide, dichloromethane, or N-methylpyrrolidone, preferably N,N-dimethylformamide. The alkaline conditions used in step (1) are potassium carbonate, cesium carbonate, or triethylamine, preferably cesium carbonate, with potassium iodide as the catalyst. The reaction temperature in step (1) is 10℃~30℃, preferably 15℃~25℃.

[0019] The solvent used in step (2) is an organic solvent, such as tetrahydrofuran, N,N-dimethylformamide, dichloromethane, or triethylamine, preferably N,N-dimethylformamide. The condensing agent used in the reaction is 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethylurea hexafluorophosphate (HATU), benzotriazole-N,N,N′,N′-tetramethylurea hexafluorophosphate (HBTU), 1-butylphosphonic anhydride (T4P), O-benzotriazole-N,N,N′,N′-tetramethylurea tetrafluoroborate (TBTU), tetramethylchlorourea hexafluorophosphate (TCFH), or carbonyl diimidazole (C... The reaction mixture is DI or (1-cyano-2-ethoxy-2-oxoethyleneaminooxy)dimethylamino-morpholine-carbomony hexafluorophosphate (COMU), preferably COMU; the base for the reaction is potassium carbonate, cesium carbonate, triethylamine, N,N-diisopropylethylamine (DIPEA), or N-methylimidazole (NMI), preferably N,N-diisopropylethylamine (DIPEA) and N-methylimidazole (NMI), preferably N-methylimidazole (NMI). The reaction temperature is 30°C to 60°C, preferably 45°C to 55°C.

[0020] The application of the paldoluno derivatives described in this invention in the treatment of tumors.

[0021] The tumors mentioned are cervical cancer, breast cancer, and kidney cancer.

[0022] Beneficial effects: Compared with the prior art, the paldoluno derivative of the present invention has good anti-tumor activity. Detailed Implementation

[0023] The technical solution of the present invention will be further described below with reference to the embodiments.

[0024] Example 1

[0025] 2-(2,6-dioxopiperidin-3-yl)-4-((2-oxo-2-(4-(2-oxo-2,3-dihydrobenzo[d]oxazol-7-yl)piperazin-1-yl)ethyl)amino)-4,5-dihydro-1H-isoindole-1,3(2H)-dione, with the following structural formula:

[0026]

[0027] Its preparation method includes the following steps:

[0028] (1) Preparation of 3-(tert-butoxycarbonylmethyl)amino-N-(2,6-dioxo-3-piperidinyl)phthalimide

[0029]

[0030] Pomalidomide 2000 mg (7.32 mmol), tert-butyl bromoacetate 1999 mg (10.25 mmol), potassium iodide 608 mg (3.66 mmol), and cesium carbonate 4772 mg (14.64 mmol) were added sequentially to a reaction flask. Acetonitrile 20 mL was used as the reaction solvent, and the reaction was carried out at 25–35 °C for 5 h. 40 mL of water was added to the reaction solution, and after stirring until homogeneous, the mixture was allowed to stand to crystallize. The crystals were filtered, washed with ethyl acetate, and dried to obtain 1733 mg, with a yield of 59.2%.

[0031] (2) Synthesis of 3-((2-(2,6-dioxo-3-piperidinyl)-1,3-dioxoisoindol-4-yl)amino)acetic acid

[0032]

[0033] The solid obtained in the previous step was added to 10 mL of ethyl acetate and hydrochloric acid gas, stirred at 15℃~25℃ for 2 h, concentrated under reduced pressure to remove excess solvent, and dried to obtain 1463 mg of dark green target product, with a yield of 98.2%.

[0034] (3) Synthesis of 2-(2,6-dioxopiridine-3-yl)-4-((2-oxo-2-(4-(2-oxo-2,3-dihydrobenzo[d]oxazol-7-yl)piperazin-1-yl)ethyl)amino)-4,5-dihydro-1H-isoindole-1,3(2H)-dione

[0035] 200 mg (0.60 mmol) of 3-((2-(2,6-dioxo-3-piperidinyl)-1,3-dioxoisoindol-4-yl)amino)propionic acid, 259 mg (0.60 mmol) of COMU, 153 mg (1.51 mmol) of NMM, and 110 mg (0.50 mmol) of 7-(piperazin-1-yl)benzo[d]oxazol-2(3H)-one were added sequentially to a reaction flask, and stirred at 45-55 °C for 3 h with 5 mL of acetonitrile as solvent. The mixture was concentrated under reduced pressure and purified by silica gel column chromatography (dichloromethane:methanol = 80:1) to give 30 mg of a yellow solid, yield 6.2%.

[0036] The target compound is a yellow solid; 1 H NMR (400MHz, DMSO-d6) δ11.58 (s, 1H), 7.48 (t, J=7.7Hz, 1H), 7.03 (q, J=7.5, 6.9Hz, 3H), 6.66 (dd, J=14.2, 8.1Hz, 2H), 6.54 (s, 2H), 5.15 (d, J=18.3Hz, 1H), 4.58 (q, J=16.1Hz, 2H), 3.66 (d, J=28.6Hz, 4H), 3.13 (m, 4H), 2.92-2.59 (m, 2H), 2.44-1.83 (m, 2H). 13 C NMR (101MHz, DMSO-d6) δ171.11, 169.21, 168.33, 167.15, 164.04, 153.77, 146.59, 135.33, 134.67, 133.32, 131.78, 130.96, 124.18, 121.56, 110.81, 109.98, 108.31, 102.67, 49.03, 48.92, 48.49, 43.78, 41.18, 40.93, 30.98, 21.06.

[0037] Example 2

[0038] The 7-piperazine-benzoxazolone compound of the present invention, chemically named 2-(2,6-dioxopiperidin-3-yl)-4-((3-oxo-3-(4-(2-oxo-2,3-dihydrobenzo[d]oxazol-7-yl)piperazin-1-yl)propyl)amino)-4,5-dihydro-1H-isoindole-1,3(2H)-dione, has the following structural formula:

[0039]

[0040] Preparation method: 125 mg (0.37 mmol) of 3-((2-(2,6-dioxadiazin-3-yl)-1,3-dioxoisoindoline-4-yl)amino)propionic acid (steps 1 and 2 are the same as in Example 1, except that tert-butyl bromoacetate in step 1 is replaced with tert-butyl bromopropionate), 100 mg (0.46 mmol) of 7-(piperazin-1-yl)benzo[d]oxazol-2(3H)-one, 234 mg (0.55 mmol) of COMU, and 138.69 mg (1.37 mmol) of NMM were reacted sequentially with 5 mL of acetonitrile as the reaction solvent. The mixture was stirred at 45℃~55℃ for 3 h. The mixture was concentrated under reduced pressure and purified by silica gel column chromatography (dichloromethane:methanol = 70:1) to obtain 33 mg of a yellow solid, with a yield of 16.6%.

[0041] The target compound is a yellow solid; 1 H NMR (400MHz, DMSO-d6) δ11.55 (s, 1H), 7.48 (dd, J=8.5, 7.0Hz, 1H), 7.03 (td, J=7.4, 6.7 , 2.5Hz, 3H), 6.65 (dd, J=17.7, 8.0Hz, 2H), 6.54 (s, 2H), 5.12 (dd, J=13.0, 5.4Hz, 1H), 3 .89 (t, J=7.9Hz, 2H), 3.60 (dt, J=13.8, 5.3Hz, 4H), 3.16 (dt, JJ=25.9, 5.3Hz, 4H), 3.02 -2.73 (m, 2H), 2.63-2.53 (m, 2H), 2.50-2.36 (m, 1H), 2.06 (dtd, J=13.0, 5.3, 2.6Hz, 1H). 13 C NMR(101MHz, DMSO)δ 172.03, 170.08, 169.01, 168.83, 167.82, 154.44, 147.24, 135.98, 135.40, 133.96, 132.44, 131.59, 124.80 , 122.24, 111.52, 110.63, 108.96, 103.24, 55.36, 49.60, 49.09, 45.12, 41.17, 36.70, 31.73, 31.09, 21.84.

[0042] Example 3

[0043] The 7-piperazine-benzoxazolone compound of the present invention, chemically named 2-(2,6-dioxopiperidin-3-yl)-4-((4-oxo-4-(4-(2-oxo-2,3-dihydrobenzo[d]oxazol-7-yl)piperazine-1-yl)butyl)amino)-4,5-dihydro-1H-isoindole-1,3(2H)-dione, has the following structural formula:

[0044]

[0045] Preparation method: 150 mg (0.41 mmol) of 3-((2-(2,6-dioxadiazin-3-yl)-1,3-dioxoisoindoline-4-yl)amino)butyric acid (steps 1 and 2 are the same as in Example 1, except that tert-butyl bromoacetate in step 1 is replaced with tert-butyl bromobutyrate), 100 mg (0.46 mmol) of 7-(piperazin-1-yl)benzo[d]oxazol-2(3H)-one, 234 mg (0.55 mmol) of COMU, and 138.69 mg (1.37 mmol) of NMM were reacted sequentially with 5 mL of acetonitrile as the reaction solvent. The mixture was stirred at 45℃~55℃ for 3 h. The mixture was concentrated under reduced pressure and purified by column chromatography (elution with dichloromethane:methanol = 70:1) to obtain 71 mg of a yellow solid, with a yield of 30.1%.

[0046] The target compound is a yellow-green solid; 1 H NMR(400MHz, DMSO-d6)δ 11.54 (s, 1H), 7.46 (t, J=7.8Hz, 1H), 7.05-6.93 (m, 3H), 6.72-6.33 (m, 4H), 5.13 (dd, J=12.9, 5.5Hz, 1H), 3.86-3.53 ( m, 6H), 3.21-3.00 (m, 4H), 3.00-2.70 (m, 2H), 2.33 (qt, J=16.0, 7.2Hz, 2H), 2.03 (p, J=5.4Hz, 2H), 1.79-1.58 (m, 2H). 13 C NMR (101MHz, DMSO-d6) δ172.17, 170.41, 169.05, 167.85, 154.44, 147.23, 147.14, 135.95, 135.45, 133.93, 132.44, 131.56 , 124.77, 122.25, 111.50, 110.42, 108.98, 108.95, 103.17, 49.54, 49.38, 49.33, 45.06, 41.34, 31.67, 29.66, 23.60, 21.88.

[0047] Example 4

[0048] The 7-piperazine-benzoxazolone compound of the present invention, chemically named 2-(2,6-dioxopiperidin-3-yl)-4-((5-oxo-5-(4-(2-oxo-2,3-dihydrobenzo[d]oxazol-7-yl)piperazin-1-yl)pentyl)amino)-4,5-dihydro-1H-isoindole-1,3(2H)-dione, has the following structural formula:

[0049]

[0050] Preparation method: 125 mg (0.37 mmol) of 3-((2-(2,6-dioxadiazin-3-yl)-1,3-dioxoisoindoline-4-yl)amino)valerate (steps 1 and 2 are the same as in Example 1, except that tert-butyl bromoacetate in step 1 is replaced with tert-butyl bromovalerate), 100 mg (0.46 mmol) of 7-(piperazin-1-yl)benzo[d]oxazol-2(3H)-one, 234 mg (0.55 mmol) of COMU, and 138.69 mg (1.37 mmol) of NMM were reacted sequentially with 5 mL of acetonitrile as the reaction solvent. The mixture was stirred at 45℃~55℃ for 3 h. The mixture was concentrated under reduced pressure and purified by column chromatography (elution with dichloromethane:methanol = 70:1) to obtain 86 mg of a yellow solid, with a yield of 28.6%.

[0051] The target compound is a yellow-green solid; 1 H NMR(400MHz, DMSO-d6)δ 7.46 (dd, J=8.5, 7.0Hz, 1H), 7.06-6.95 (m, 3H), 6.60 (dd, J=28.2, 19.9Hz, 4H), 5.13 (dd, J=12.9, 5.4Hz, 1H), 3.70-3.60 (m, 6H), 3. 13 (dt, J==21.1, 5.5Hz, 4H), 2.99-2.70 (m, 2H), 2.36 (q, J=7.0, 5.6Hz, 2H), 2.03 (dtd, J=10.8, 5.3, 2.5Hz, 2H), 1.51-1.46 (m, 4H). 13 C NMR(101MHz, DMSO-d6)δ 172.12, 170.95, 170.10, 169.04, 167.83, 154.44, 147.22, 135.92, 135.41, 133.95, 132.42, 131.64, 124.77, 12 2.22, 111.47, 110.53, 108.95, 103.27, 49.66, 49.56, 49.27, 45.24, 41.28, 32.31, 31.66, 27.64, 22.54, 21.93.

[0052] Example 5

[0053] The 7-piperazine-benzoxazolone compound of the present invention, chemically named 2-(2,6-dioxopiridin-3-yl)-4-((6-oxo-6-(4-(2-oxo-2,3-dihydrobenzo[d]oxazol-7-yl)piperazine-1-yl)hexyl)amino)-4,5-dihydro-1H-isoindole-1,3(2H)-dione, has the following structural formula, and its preparation method includes the following steps:

[0054]

[0055] Preparation method: 200 mg (0.48 mmol) of 3-((2-(2,6-dioxadiazin-3-yl)-1,3-dioxoisoindoline-4-yl)amino)hexanoic acid (steps 1 and 2 are the same as in Example 1, except that tert-butyl bromoacetate in step 1 is replaced with tert-butyl bromohexanoate), 122 mg (0.46 mmol) of 7-(piperazin-1-yl)benzo[d]oxazol-2(3H)-one, 234 mg (0.55 mmol) of COMU, and 139 mg (1.37 mmol) of NMM were reacted sequentially with 5 mL of acetonitrile as the reaction solvent. The mixture was stirred at 45℃~55℃ for 3 h. The mixture was concentrated under reduced pressure and purified by silica gel column chromatography (dichloromethane:methanol = 70:1) to obtain 102 mg of a yellow solid, with a yield of 36.2%.

[0056] The target compound is a yellow solid; 1 H NMR (400MHz, DMSO-d6) δ11.57 (s, 1H), 7.46 (dd, J = 8.5, 7.0Hz, 1H), 7.08-6.97 (m, 3H), 6.73 -6.49 (m, 4H), 5.13 (dd, J=12.9, 5.5Hz, 1H), 3.81-3.65 (m, 2H), 3.61 (td, J=7.3, 3.4Hz, 4H), 3.20-3.08 (m, 4H), 2.98 (ddd, J=17.0, 14.0, 5.3Hz, 2H), 2.74 (ddd, J=17.1, 4.4, 2.7Hz, 1H), 2.38-2.29 (m, 2H), 2.07-1.99 (m, 1H), 1.58-1.40 (m, 4H), 1.27 (td, J=10.5, 9.8, 5.2Hz, 2H). 13C NMR(101MHz,DMSO-d6)δ 172.08, 171.08, 170.10, 169.03, 167.82, 154.43, 147.23, 135.93, 135.45, 133.96, 132.46, 131.59, 124.78, 122.21, 1 11.46, 110.56, 108.98, 103.21, 55.37, 49.57, 49.30, 49.07, 45.25, 41.24, 32.56, 31.66, 27.64, 26.36, 24.93, 21.94.

[0057] Bioactivity evaluation

[0058] MTT assay for antitumor activity

[0059] Cell culture: Human cancer cell lines MDA-MB-231 and HeLa cells were purchased from the National Biomedical Laboratory in Beijing and cultured in DMEM (KGM12800-500) or MEM (KGM41500-500) medium containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin, at 37°C in a Thermo Fisher Scientific (BB150) incubator containing 5% CO2. When the cell confluence reached 70%-80%, 0.25% trypsin was added for digestion, resuspending, and culturing. Cells in the logarithmic growth phase and in good growth condition were selected for study.

[0060] Methyl thiazolyl tetrazolium (MTT) was used to determine cell viability. Hemocytometer counting was used for cell counting, and cell viability was greater than 95% in all experiments. MDA-MB-231 and HeLa cells were counted at 1 × 10⁻⁶ cells / mL. 4 Cells were seeded in 96-well plates. 100 μL of medium (containing 1% FBS) was added to each well to dissolve different concentrations of the drug (0-50 μM), and the cells were incubated for 24 h. After centrifugation (5 min, 2000 rpm), the supernatant was discarded, and 10 μL of LTT (5 mg / mL) solution was added to each well. The cells were incubated at 37°C for 4 h, centrifuged again, and the supernatant was discarded. 100 μL of DMSO was added to each well, and the cells were shaken for 10 min to fully dissolve the formazan crystals. The absorbance was measured at 570 nm using a microplate reader (BioTek, USA). Cytotoxicity was assessed compared to the control group (DMSO). The concentration at which the drug induced 50% cell growth inhibition (IC50) was determined using a curve fitting algorithm in GraphPad Prism9 (GraphPad software, La Jolla, CA, USA) via nonlinear regression.50 ).

[0061] To investigate the effects of the compounds on tumor cell activity, cytotoxicity was detected using the MTT assay. Table 1 shows the effects of the compounds on the activity of different tumor cell lines, and the half-maximal inhibitory concentration (IC50) of the drugs was calculated for each cell line. s0 (48h).

[0062] Table 1. Inhibitory activity of compounds against different tumor cells.

[0063] Example MDA-MB-231(μM) Hela (μM) Example 1 165.77±6.62 92.38±2.03 Example 2 126.23±3.96 / Example 3 90.71±1.62 44.11±2.01 Example 4 94.82±17.92 / Example 5 187.4±8.66 24.19±2.34 7-Piperazine-Benzoxazolone 222.47±30.69 74.68±4.81 pomalidomide 389.47+35.83 572.73±13.28

[0064] " / " indicates that it was not measured.

[0065] The results in Table 1 show that the compound has a significant inhibitory effect on MDA-MB-231 and HeLa tumor cells, and has great research value.

Claims

1. A compound of formula (I), or a salt thereof, or a hydrate thereof: in, n is 1 to 5.

2. The pardoprunox derivative according to claim 1, characterized in that, It is a compound or a pharmaceutically acceptable salt thereof, as shown below:

3. A method for preparing the paldoluno derivative of claim 1, characterized in that, The following steps are included: Compound 4 and Compound 5 are amidated to obtain Compound I:

4. A pharmaceutical composition, characterized in that, It includes the compounds as described in claims 1 to 2 and at least one pharmaceutically acceptable excipient.

5. The use of a substance in the preparation of a medicament for treating diseases, characterized in that, The substance is a compound as described in claims 1-2, and the disease is cancer; preferably cervical cancer, breast cancer, and kidney cancer.

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