A class of proteolysis-targeting chimeric compounds targeting the degradation of GSK-3β and their applications

Abstract

The present invention discloses a class of proteolysis-targeting chimeric compounds targeting the degradation of GSK-3β and their applications. The compounds have a general structural formula as shown in Formula P: wherein: R is selected from one of the following: hydrogen, halogen, alkyl with 1-5 carbon atoms, alkoxy with 1-5 carbon atoms; n = 1-7. The compounds provided by the present invention have good inhibitory activity against GSK-3β and can be used for the treatment of GSK-3β-targeted related diseases. The present invention tested the GSK-3β inhibitory activity of 9 target compounds P1 to P9 synthesized, and the results showed that they all exhibited good inhibitory activity against GSK-3β, and the IC 50 value was 1.41 ± 0.29 to 5.46 ± 0.25 μM.

Description

Technical Field

[0001] The present invention mainly relates to a class of proteolysis-targeting chimera compounds that target the degradation of GSK-3β and their applications. Background Art

[0002] Glycogen synthase kinase 3 (GSK-3) belongs to a multifunctional serine protein kinase in the phosphotransferase family. There are two subtypes of mammalian GSK-3, namely GSK-3α and GSK-3β, which have a high degree of homology in their catalytic domains, but there are significant differences in the C-terminal and N-terminal domains. GSK-3α and GSK-3β have only 36% sequence identity at their carboxyl termini, and GSK-3α has a glycine-rich extension at its amino terminus. GSK-3β is highly abundant in the brain, mainly concentrated in neurons and astrocytes, and its expression level increases with age. GSK-3β is a therapeutic target for many diseases. GSK3β participates in regulating glucose metabolism, cell signaling, cell proliferation, growth, migration, differentiation, cell cycle, embryonic development, apoptosis, insulin response, and various transcription factors to regulate the growth and death of organs. GSK3β plays a role in many diseases, such as diabetes, Alzheimer's disease inflammation, cancer, etc. Inhibiting the activity of GSK3β is of great significance for the treatment of the above diseases. Developing proteolysis-targeting chimera compounds that target the degradation of GSK-3β can effectively reduce the expression of GSK-3β in cells, thereby effectively treating related diseases targeted by GSK-3β. Summary of the Invention

[0003] Aiming at the above technical problems existing in the prior art, the purpose of the present invention is to provide a class of proteolysis-targeting chimera compounds that target the degradation of GSK-3β and their applications. The compounds of the present invention are used to treat related diseases.

[0004] The technical solutions obtained by the present invention are as follows:

[0005] A class of proteolysis-targeting chimera compounds that target the degradation of GSK-3β, and this class of compounds has a general structural formula as shown in Formula P:

[0006]

[0007] Wherein: R is selected from one of the following: hydrogen, halogen, an alkyl group with 1-5 carbon atoms, an alkoxy group with 1-5 carbon atoms; n = 1-7.

[0008] The present invention also provides a synthesis route for the above-mentioned class of proteolysis-targeting chimera compounds that target the degradation of GSK-3β, specifically as follows:

[0009]

[0010] (1) First, using N-phenylmaleimide as a raw material, it is chlorinated with thionyl chloride to obtain Compound 2, and then in the presence of ethylmagnesium bromide, it is condensed with indole Grignard reagent at room temperature to obtain Compound 3;

[0011] (2) Using copper oxide as a catalyst, Compound 3 undergoes N-methylation reaction with methyl iodide under the action of potassium carbonate to obtain Compound 4;

[0012] (3) Using anhydrous benzene-THF with a volume ratio of 1 - 1.2:1 as a solvent, in the presence of ethylmagnesium bromide, Compound 4 is condensed with indole Grignard reagent with substituent R at reflux to obtain Compound 5;

[0013] (4) Compound 5 undergoes N-alkylation reaction with ethyl bromoacetate under the action of cesium carbonate to obtain Compound 6;

[0014] (5) Compound 6 undergoes hydrolysis reaction under alkaline conditions to obtain Compound 7;

[0015] (6) Compound 7 undergoes ammonolysis reaction with molten ammonium acetate under nitrogen protection to obtain Compound 8;

[0016] The substituents R on the benzene rings of Compounds 5 - 8 and the indole Grignard reagent described in step (3) are the same as those in Formula P.

[0017] (7) Compound 9 undergoes condensation reaction with Compound 10 under the action of DIPEA to obtain Compound 11;

[0018] (8) Compound 11 undergoes hydrogenation reduction reaction with H2 under the catalysis of palladium on carbon to obtain Compound 12;

[0019] The n in Compounds 10 - 12 is the same as that in Formula P.

[0020] (9) Compound 8 and Compound 12 undergo condensation reaction in the presence of TEA and HATU to obtain the target product.

[0021] The present invention also provides the application of the class of proteolysis-targeting chimeric compounds targeting the degradation of GSK-3β in the preparation of drugs for regulating the GSK-3β signaling pathway.

[0022] By adopting the above technologies, compared with the prior art, the beneficial effects of the present invention are as follows:

[0023] The proteolysis-targeting chimeric compounds targeting the degradation of GSK-3β designed and synthesized by the present invention are a class of novel compounds and can be used for the treatment of GSK-3β-targeted related diseases. Detailed implementation mode

[0024] The present invention will be further described below in conjunction with specific embodiments, but the protection scope of the present invention is not limited thereto.

[0025] 1. Preparation of intermediates and target compounds:

[0026] Example 1: N-phenyldichloromaleimide (2)

[0027]

[0028] Add 22.5 g (0.13 mol) of N-phenylmaleimide and 160 mL of thionyl chloride into a three-necked flask. After stirring and dissolving, cool to 0 °C, slowly add dropwise 25 mL of anhydrous pyridine, and reflux for 2 h after addition. After the reaction is completed, concentrate under reduced pressure to remove the excessive thionyl chloride. Pour the residue into 200 mL of ice water, extract with dichloromethane (100 mL × 3), combine the organic phases, wash the organic phases with saturated brine (300 mL × 3), and dry over anhydrous sodium sulfate. Filter, concentrate the filtrate under reduced pressure, recrystallize with 100 mL of dichloromethane, filter, and dry to obtain 20.0 g of yellow solid compound 2 with a yield of 60.0%.

[0029] Example 2: 3-chloro-4-(1H-indol-3-yl)-1-phenyl-1H-pyrrole-2,5-dione (3)

[0030]

[0031] Add 3.0 g (25.74 mmol) of indole and 5 mL of anhydrous benzene into a three-necked flask. Under nitrogen protection, add dropwise 12.5 mL (25.00 mmol) of ethylmagnesium bromide solution (2 mol / L in THF). After the addition, react at room temperature for 30 min to obtain indole Grignard reagent. Add 4.0 g (16.50 mmol) of compound 2 into the reaction flask, protect with nitrogen, inject 10 mL of THF, and slowly add dropwise the indole Grignard reagent at low temperature. After the addition, react at room temperature for 4 h. After the reaction is completed, pour into 100 mL of ice water to quench, adjust to weakly acidic with hydrochloric acid aqueous solution, extract with dichloromethane (50 mL × 3), combine the organic phases, wash the organic phases with saturated brine (150 mL × 3), and dry over anhydrous sodium sulfate. Filter, concentrate the filtrate under reduced pressure, recrystallize with ethyl acetate, filter, and dry to obtain 2.9 g of red solid compound 3 with a yield of 54.4%.

[0032] Example 3: 3-chloro-4-(1-methyl-1H-indol-3-yl)-1-phenyl-1H-pyrrole-2,5-dione (4)

[0033]

[0034] 500 mg (1.55 mmol) of compound 3, 10 mL of DMF, 214 mg (1.55 mmol) of potassium carbonate, 99 mg (1.24 mmol) of copper oxide, and 8.8 g (62.00 mmol) of methyl iodide were added to a reaction flask, and the reaction was carried out at room temperature for 4 h. After the reaction was completed, it was poured into 100 mL of water for quenching, and extracted with dichloromethane (50 mL×3). The organic phases were combined, washed with saturated brine (150 mL×3), and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, recrystallized from ethyl acetate, filtered, and dried to obtain 250 mg of red solid compound 4 with a yield of 47.9%.

[0035] Example 4: 3-(1H-Indol-3-yl)-4-(1-methyl-1H-indol-3-yl)-1-phenyl-1H-pyrrole-2,5-dione (5a)

[0036]

[0037] 1.1 g (9.28 mmol) of indole and 5 mL of anhydrous benzene were added to a three-necked flask under nitrogen protection. 4.46 mL (8.92 mmol) of ethylmagnesium bromide solution (2 mol / L in THF) was slowly added dropwise at low temperature. After the addition, the reaction was carried out at room temperature for 30 min to obtain indole Grignard reagent (for later use). 2.0 g (5.95 mmol) of compound 4 was added to the reaction flask under nitrogen protection, 10 mL of THF was injected, and the indole Grignard reagent was slowly added dropwise at low temperature. After the addition, the reaction was refluxed for 4 h. After the reaction was completed, it was poured into ice water for quenching, adjusted to weakly acidic with 2N hydrochloric acid solution, extracted with dichloromethane (50 mL×3). The organic phases were combined, washed with saturated brine (150 mL×3), and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM:PE = 10:1, v / v) to obtain 1.5 g of red solid 5a with a yield of 60.5%.

[0038] Example 5: 3-(4-Bromo-1H-indol-3-yl)-4-(1-methyl-1H-indol-3-yl)-1-phenyl-1H-pyrrole-2,5-dione (5b)

[0039]

[0040] The synthesis procedure of compound 5b was the same as that of compound 5a, except that indole was replaced with an equimolar amount of 4-bromoindole, and the reaction gave a red solid 5b with a yield of 11.9%.

[0041] Example 6: 3-(5-Bromo-1H-indol-3-yl)-4-(limethyl-1H-indol-3-yl)-1-phenyl-1H-pyrrole-2,5-dione (5c)

[0042]

[0043] The synthesis procedure of compound 5c is the same as that of compound 5a, except that indole is replaced with an equimolar amount of 5-bromoindole, and the reaction gives a red solid 5c with a yield of 20.3%.

[0044] Example 7: 3-(6-Bromo-1H-indol-3-yl)-4-(1-methyl-1H-indol-3-yl)-1-phenyl-1H-pyrrole-2,5-dione (5d)

[0045]

[0046] The synthesis procedure of compound 5d is the same as that of compound 5a, except that indole is replaced with an equimolar amount of 6-bromoindole, and the reaction gives a red solid 5d with a yield of 67.7%.

[0047] Example 8: 3-(5-Methoxy-1H-indol-3-yl)-4-(1-methyl-1H-indol-3-yl)-1-phenyl-1H-pyrrole-2,5-dione (5e)

[0048]

[0049] The synthesis procedure of compound 5e is the same as that of compound 5a, except that indole is replaced with an equimolar amount of 5-methoxyindole, and the reaction gives a red solid 5e with a yield of 56.3%.

[0050] Example 9: Ethyl 2-(3-(4-(1-methyl-1H-indol-3-yl)-2,5-dioxo-1-phenyl-2,5-dihydro-1H-pyrrol-3-yl)-1H-indol-1-yl)acetate (6a)

[0051]

[0052] Add 100 mg (0.24 mmol) of compound 5a, 5 mL of DMF, and 234 mg (0.72 mmol) of cesium carbonate into a reaction flask, react at room temperature for 15 min, then add 80 mg (0.48 mmol) of ethyl bromoacetate and react at room temperature for 1 h. After the reaction is completed, pour it into 25 mL of water to quench, extract with ethyl acetate (20 mL × 3), combine the organic phases, wash the organic phases with saturated brine (50 mL × 3), and dry over anhydrous sodium sulfate. Filter, concentrate the filtrate under reduced pressure, and purify the residue by silica gel column chromatography (DCM:PE:EA = 1:2:1, v / v / v) to obtain 110 mg of a red solid 6a with a yield of 88.4%.

[0053] Example 10: 2-(4-bromo-3-(4-(1-methyl-1H-indol-3-yl)-2,5-dioxo-1-phenyl-2,5-dihydro-1H-pyrrol-3-yl)ethyl)-1H-indol-1-yl) acetate (6b)

[0054]

[0055] The synthesis procedure of compound 6b is the same as that of compound 6a, except that compound 5a is replaced with an equimolar amount of compound 5b, and the reaction gives a red solid 6b with a yield of 78.1%.

[0056] Example 11: 2-(5-bromo-3-(4-(1-methyl-1H-indol-3-yl)-2,5-dioxo-1-phenyl-2,5-dihydro-1H-pyrrol-3-yl)ethyl)-1H-indol-1-yl) acetate (6c)

[0057]

[0058] The synthesis procedure of compound 6c is the same as that of compound 6a, except that compound 5a is replaced with an equimolar amount of compound 5c, and the reaction gives a red solid 6c with a yield of 87.6%.

[0059] Example 12: 2-(6-bromo-3-(4-(1-methyl-1H-indol-3-yl)-2,5-dioxo-1-phenyl-2,5-dihydro-1H-pyrrol-3-yl)ethyl)-1H-indol-1-yl) acetate (6d)

[0060]

[0061] The synthesis procedure of compound 6d is the same as that of compound 6a, except that compound 5a is replaced with an equimolar amount of compound 5d, and the reaction gives a red solid 6d with a yield of 82.0%.

[0062] Example 13: 2-(5-methoxy-3-(4-(1-methyl-1H-indol-3-yl)-2,5-dioxo-1-phenyl-2,5-dihydro-1H-pyrrol-3-yl)ethyl)-1H-indol-1-yl) acetate (6e)

[0063]

[0064] The synthesis procedure of compound 6e is the same as that of compound 6a, except that compound 5a is replaced with an equimolar amount of compound 5e, and the reaction gives a red solid 6e with a yield of 73.5%.

[0065] Example 14: 2-(3-(4-(1-Methyl-1H-indol-3-yl)-2,5-dioxo-2,5-dihydrofuran-3-yl)-1H-indol-1-yl)acetic acid (7a)

[0066]

[0067] Add 110 mg (0.21 mmol) of compound 6a and 2.5 mL of THF to a reaction flask. Dissolve 25.2 mg (1.05 mmol) of lithium hydroxide in 2.5 mL of water, then add the lithium hydroxide aqueous solution to the reaction flask. Add 0.5 mL of methanol and react at room temperature for 1 h. After the reaction is completed, pour it into an appropriate amount of water to quench, adjust to acidic with hydrochloric acid aqueous solution, extract with ethyl acetate (20 mL × 3), combine the organic phases, wash the organic phases with saturated brine (50 mL × 3), and dry over anhydrous sodium sulfate. Filter, concentrate the filtrate under reduced pressure to obtain 100 mg of red solid 7a, which is directly used for the next step.

[0068] Example 15: 2-(4-Bromo-3-(4-(1-methyl-1H-indol-3-yl)-2,5-dioxo-2,5-dihydrofuran-3-yl)-1H-indol-1-yl)acetic acid (7b)

[0069]

[0070] The synthesis procedure of compound 7b is the same as that of compound 7a, except that compound 6a is replaced with an equimolar amount of compound 6b, and the reaction gives red solid 7b, which is directly used for the next step.

[0071] Example 16: 2-(5-Bromo-3-(4-(1-methyl-1H-indol-3-yl)-2,5-dioxo-2,5-dihydrofuran-3-yl)-1H-indol-1-yl)acetic acid (7c)

[0072]

[0073] The synthesis procedure of compound 7c is the same as that of compound 7a, except that compound 6a is replaced with an equimolar amount of compound 6c, and the reaction gives red solid 7c, which is directly used for the next step.

[0074] Example 17: 2-(6-Bromo-3-(4-(1-methyl-1H-indol-3-yl)-2,5-dioxo-2,5-dihydrofuran-3-yl)-1H-indol-1-yl)acetic acid (7d)

[0075]

[0076] The synthesis procedure of compound 7d is the same as that of compound 7a, except that compound 6a is replaced with an equimolar amount of compound 6d. The reaction gives a red solid 7d, which is directly used in the next step.

[0077] Example 18: 2-(5-Methoxy-3-(4-(1-methyl-1H-indol-3-yl)-2,5-dioxo-2,5-dihydrofuran-3-yl)-1H-indol-1-yl)acetic acid (7e)

[0078]

[0079] The synthesis procedure of compound 7e is the same as that of compound 7a, except that compound 6a is replaced with an equimolar amount of compound 6e. The reaction gives a red solid 7e, which is directly used in the next step.

[0080] Example 19: 2-(3-(4-(1-methyl-1H-indol-3-yl)-2,5-dioxo-2,5-dihydro-1H-pyrrol-3-yl)-1H-indol-1-yl)acetic acid (8a)

[0081]

[0082] Add 0.25 mmol of compound 7a and 2.0 g (25 mmol) of ammonium acetate into a reaction flask, protect with nitrogen, and react at 140 °C for 5 h. After the reaction is completed, pour it into 50 mL of water to quench, extract with ethyl acetate (50 mL × 3), combine the organic phases, wash the organic phases with saturated brine (100 mL × 3), and dry over anhydrous sodium sulfate. Filter, concentrate the filtrate under reduced pressure, and purify the residue by silica gel column chromatography (PE:EA:AcOH = 50:50:1, v / v / v) to obtain 50 mg of red solid 8a, with a two-step yield of 50.1%.

[0083] Example 20: 2-(4-Bromo-3-(4-(1-methyl-1H-indol-3-yl)-2,5-dioxo-2,5-dihydro-1H-pyrrol-3-yl)-1H-indol-1-yl)acetic acid (8b)

[0084]

[0085] The synthesis procedure of compound 8b is the same as that of compound 8a, except that compound 7a is replaced with an equimolar amount of compound 7b. The reaction gives a red solid 8b, with a two-step yield of 22.1%.

[0086] Example 21: 2-(5-Bromo-3-(4-(1-methyl-1H-indol-3-yl)-2,5-dioxo-2,5-dihydro-1H-pyrrol-3-yl)-1H-indol-1-yl)acetic acid (8c)

[0087]

[0088] The synthesis procedure of compound 8c is the same as that of compound 8a, except that compound 7a is replaced with an equimolar amount of compound 7c, and the reaction gives a red solid 8c with a two-step yield of 31.3%.

[0089] Example 22: 2-(6-Bromo-3-(4-(1-methyl-1H-indol-3-yl)-2,5-dioxo-2,5-dihydro-1H-pyrrol-3-yl)-1H-indol-1-yl)acetic acid (8d)

[0090]

[0091] The synthesis procedure of compound 8d is the same as that of compound 8a, except that compound 7a is replaced with an equimolar amount of compound 7d, and the reaction gives a red solid 8d with a two-step yield of 21.9%.

[0092] Example 23: 2-(5-Methoxy-3-(4-(1-methyl-1H-indol-3-yl)-2,5-dioxo-2,5-dihydro-1H-pyrrol-3-yl)-1H-indol-1-yl)acetic acid (8e)

[0093]

[0094] The synthesis procedure of compound 8e is the same as that of compound 8a, except that compound 7a is replaced with an equimolar amount of compound 7e, and the reaction gives a red solid 8e with a two-step yield of 23.4%.

[0095] Example 24: 4-((2-(2-(2-(2-Azidoethoxy)ethoxy)ethoxy)ethyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (11a)

[0096]

[0097] Add 293 mg (1.06 mmol) of compound 9, 300 mg (1.38 mmol) of compound 10a, 10 mL of DMF, and 357 mg (2.76 mmol) of N,N-diisopropylethylamine to a reaction flask, and heat to 80 °C for reaction for 4 h. After the reaction is completed, pour it into 100 mL of ice water to quench, extract with ethyl acetate (50 mL × 3), combine the organic phases, wash the organic phases with saturated brine (150 mL × 3), and dry over anhydrous sodium sulfate. Filter, concentrate the filtrate under reduced pressure, and purify the residue by silica gel column chromatography (PE:EA = 1:4, v / v) to obtain 120 mg of a green oily liquid 11a with a yield of 23.8%.

[0098] In the molecular structure of Compound 10a, n = 3.

[0099] Example 25: 4-((14-Azido-3,6,9,12-tetraoxatetradecyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (11b)

[0100]

[0101] The synthesis procedure of Compound 11b is the same as that of Compound 11a, except that Compound 10a is replaced with an equimolar amount of Compound 10b, and a green oily liquid 11b is obtained by the reaction, with a yield of 32.6%.

[0102] In the molecular structure of Compound 10b, n = 4.

[0103] Example 26: 4-((17-Azido-3,6,9,12,15-pentaoxaheptadecyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (11c)

[0104]

[0105] The synthesis procedure of Compound 11c is the same as that of Compound 11a, except that Compound 10a is replaced with an equimolar amount of Compound 10c, and a green oily liquid 11c is obtained by the reaction, with a yield of 38.8%.

[0106] In the molecular structure of Compound 10c, n = 5.

[0107] Example 27: 4-((20-Azido-3,6,9,12,15,18-hexaoxaeicosyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (11d)

[0108]

[0109] The synthesis procedure of Compound 11d is the same as that of Compound 11a, except that Compound 10a is replaced with an equimolar amount of Compound 10d, and a green oily liquid 11d is obtained by the reaction, with a yield of 36.7%.

[0110] In the molecular structure of Compound 10d, n = 6.

[0111] Example 28: 4-((23-Azido-3,6,9,12,15,18,21-heptaoxatricosyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (11e)

[0112]

[0113] The synthesis procedure of Compound 11e is the same as that of Compound 11a, except that Compound 10a is replaced with an equimolar amount of Compound 10e, and a green oily liquid 11e is obtained by reaction with a yield of 22.5%.

[0114] In the molecular structure of Compound 10e, n = 7.

[0115] Example 29: 4-((2-(2-(2-(2-(2-Aminoethoxy)ethoxy)ethoxy)ethyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (12a)

[0116]

[0117] 90 mg (0.19 mmol) of Compound 11a, 18 mg of palladium-carbon, and 5 mL of ethanol were added to a reaction flask, and the reaction was carried out at room temperature for 1 h. After the reaction was completed, the palladium-carbon was removed by suction filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM:MeOH:TEA = 100:10:1, v / v / v) to obtain 50 mg of a green oily liquid 12a with a yield of 85.1%.

[0118] Example 30: 4-((14-Amino-3,6,9,12-tetraoxatetradecyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (12b)

[0119]

[0120] The synthesis procedure of Compound 12b is the same as that of Compound 12a, except that Compound 11a is replaced with an equimolar amount of Compound 11b, and a green oily liquid 12b is obtained by reaction with a yield of 70.0%.

[0121] Example 31: 4-((17-Amino-3,6,9,12,15-pentaoxaheptadecyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (12c)

[0122]

[0123] The synthesis procedure of Compound 12c is the same as that of Compound 12a, except that Compound 11a is replaced with an equimolar amount of Compound 11c, and a green oily liquid 12c is obtained by reaction with a yield of 52.6%.

[0124] Example 32: 4-((20-Amino-3,6,9,12,15,18-hexaoxaeicosyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (12d)

[0125]

[0126] The synthesis procedure of compound 12d is the same as that of compound 12a, except that compound 11a is replaced with an equimolar amount of compound 11d, and a green oily liquid 12d is obtained by reaction, with a yield of 65.5%.

[0127] Example 33: 4-((23-Amino-3,6,9,12,15,18,21-heptaoxatricosyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (12e)

[0128]

[0129] The synthesis procedure of compound 12e is the same as that of compound 12a, except that compound 11a is replaced with an equimolar amount of compound 11e, and a green oily liquid 12e is obtained by reaction, with a yield of 60.9%.

[0130] Example 34: N-(2-(2-(2-(2-((2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)ethyl)-2-(3-(4-(1-methyl-1H-indol-3-yl)-2,5-dioxo-2,5-dihydro-1H-pyrrol-3-yl)-1H-indol-1-yl)acetamide (P1)

[0131]

[0132] Add 22 mg (0.055 mmol) of compound 8a, 20 mg (0.05 mmol) of compound 12a, 16 mg (0.15 mmol) of triethylamine, 5 mL of DMF, and 23 mg (0.06 mmol) of HATU into a reaction flask, and react at room temperature for 2 h. After the reaction is completed, pour an appropriate amount of water to quench the reaction, extract with ethyl acetate (20 mL × 3), combine the organic phases, wash the organic phases with saturated brine (50 mL × 3), and dry over anhydrous sodium sulfate. Filter, concentrate the filtrate under reduced pressure, and purify the residue by silica gel column chromatography (EA:MeOH = 15:1, v / v) to obtain a red solid P1 with a yield of 34.0%. 11H NMR (400 MHz, DMSO-d6) δ 11.11 (s, 1H), 10.94 (s, 1H), 8.41–8.27 (m, 1H), 7.81 (s, 2H), 7.67–7.51 (m, 1H), 7.42 (d, J = 8.2 Hz, 1H), 7.33 (d, J = 8.2 Hz, 1H), 7.13 (d, J = 8.7, 2.3 Hz, 1H), 7.10–6.96 (m, 3H), 6.79 (t, J = 8.8 Hz, 2H), 6.71–6.57 (m, 3H), 5.06 (dd, J = 12.9, 5.3 Hz, 1H), 4.92 (s, 2H), 3.85 (s, 3H), 3.67–3.59 (m, 2H), 3.59–3.40 (m, 12H), 3.30–3.22 (m, 2H), 2.95–2.82 (m, 1H), 2.66–2.52 (m, 2H), 2.06–2.00 (m, 1H). ESI-MS: m / z [M+H] + 830.

[0133] Example 35: N-(14-((2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-3,6,9,12-tetraoxatetradecyl)-2-(3-(4-(1-methyl-1H-indol-3-yl)-2,5-dioxo-2,5-dihydro-1H-pyrrol-3-yl)-1H-indol-1-yl)acetamide (P2)

[0134]

[0135] The synthesis procedure of compound P2 was the same as that of P1, except that compound 12a was replaced with an equimolar amount of compound 12b, and the reaction gave a red solid P2 with a yield of 22.8%. 1HNMR(400MHz,DMSO-d6)δ11.10(s,1H),10.93(s,1H),8.30(t,J=5.6Hz,1H),7.81(d,J=1.2Hz,2H),7.57(t,J=8.6,7.1Hz,1H),7.41(d,J=8.2Hz,1H),7.33(d,J=8.3Hz,1H),7.13(d,J=8.6Hz,1H),7.09–6.97(m,3H),6.85–6.72(m,2H),6.72–6.54(m,3H),5.06(dd,J=12.9,5.4Hz,1H),4.91(s,2H),3.85(s,3H),3.61(t,J=5.4Hz,2H),3.57–3.41(m,16H),3.27(q,J=5.7Hz,2H),2.94–2.81(m,1H),2.67–2.52(m,2H),2.09–2.01(m,1H).ESI-MS:m / z[M+H] + 874.

[0136] Example 36: N-(17-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-3,6,9,12,15-pentaoxaheptadecyl)-2-(3-(4-(1-methyl-1H-indol-3-yl)-2,5-dioxo-2,5-dihydro-1H-pyrrol-3-yl)-1H-indol-1-yl)acetamide (P3)

[0137]

[0138] The synthesis procedure of compound P3 was the same as that of P1, except that compound 12a was replaced with an equimolar amount of compound 12c, and the reaction gave a red solid P3 with a yield of 19.7%. 1HNMR(400MHz, DMSO-d6) δ 11.10 (s, 1H), 10.93 (s, 1H), 8.30 (t, J = 5.6 Hz, 1H), 7.81 (d, J = 1.4 Hz, 2H), 7.62–7.54 (m, 1H), 7.41 (d, J = 8.2, 1.0 Hz, 1H), 7.33 (d, J = 8.3 Hz, 1H), 7.13 (d, J = 8.6 Hz, 1H), 7.10–6.97 (m, 3H), 6.79 (t, J = 9.2, 8.2, 1.0 Hz, 2H), 6.70–6.56 (m, 3H), 5.06 (dd, J = 12.9, 5.4 Hz, 1H), 4.92 (s, 2H), 3.85 (s, 3H), 3.62 (d, J = 5.5 Hz, 2H), 3.58–3.42 (m, 20H), 3.27 (q, J = 5.6 Hz, 2H), 2.96–2.81 (m, 1H), 2.63–2.53 (m, 2H), 2.06–2.00 (m, 1H). ESI-MS: m / z [M-H] + 916.

[0139] Example 37: N-(20-((2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-3,6,9,12,15,18-hexaoxaeicosyl)-2-(3-(4-(1-methyl-1H-indol-3-yl)-2,5-dioxo-2,5-dihydro-1H-pyrrol-3-yl)-1H-indol-1-yl)acetamide (P4)

[0140]

[0141] The synthesis procedure of compound P4 is the same as that of P1, except that compound 12a is replaced with an equimolar amount of compound 12d, and the reaction gives a red solid P4 with a yield of 16.9%. 1HNMR(400MHz,DMSO-d6)δ11.11(s,1H),10.94(s,1H),8.32(t,J=5.6Hz,1H),7.81(d,J=1.9Hz,2H),7.57(q,J=8.6,7.1Hz,1H),7.41(d,J=8.2Hz,1H),7.33(d,J=8.3Hz,1H),7.13(d,J=8.6Hz,1H),7.08–6.95(m,3H),6.84–6.73(m,2H),6.68–6.57(m,3H),5.06(dd,J=12.9,5.4Hz,1H),4.92(s,2H),3.85(s,3H),3.61(t,J=5.4Hz,2H),3.58–3.41(m,24H),3.27(q,J=5.6Hz,2H),2.98–2.80(m,1H),2.64–2.53(m,2H),2.08–2.01(m,1H).ESI-MS:m / z[M+H] + 963.

[0142] Example 38: N-(23-((2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-3,6,9,12,15,18,21-heptaoxatricosyl)-2-(3-(4-(1-methyl-1H-indol-3-yl)-2,5-dioxo-2,5-dihydro-1H-pyrrol-3-yl)-1H-indol-1-yl)acetamide (P5)

[0143]

[0144] The synthesis procedure of compound P5 was the same as that of P1, except that compound 12a was replaced with an equimolar amount of compound 12e, and the reaction gave a red solid P5 in a yield of 13.0%. 1HNMR(400MHz,DMSO-d6)δ11.11(s,1H),10.94(s,1H),8.34(t,J=5.6Hz,1H),7.81(d,J=1.7Hz,2H),7.58(q,J=8.5,7.1Hz,1H),7.41(d,J=8.3Hz,1H),7.33(d,J=8.2Hz,1H),7.14(d,J=8.6Hz,1H),7.09–6.96(m,3H),6.78(t,J=8.7Hz,2H),6.70–6.56(m,3H),5.06(dd,J=12.9,5.4Hz,1H),4.92(s,2H),3.85(s,3H),3.61(t,J=5.4Hz,2H),3.57–3.43(m,28H),3.27(q,J=5.6Hz,2H),2.95–2.81(m,1H),2.62–2.54(m,2H),2.06–2.01(m,1H).

[0145] Example 39: 2-(4-bromo-3-(4-(1-methyl-1H-indol-3-yl)-2,5-dioxo-2,5-dihydro-1H-pyrrol-3-yl)-1H-indol-1-yl)-N-(20-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-3,6,9,12,15,18-hexaoxa)acetamide (P6)

[0146]

[0147] The synthesis procedure of compound P6 was the same as that of P1, except that compound 12a was replaced with an equimolar amount of compound 12d, compound 8a was replaced with compound 8b, and triethylamine was replaced with an equimolar amount of N,N-diisopropylethylamine. The reaction yielded a red solid P6 with a yield of 30.7%. 1HNMR(400MHz, DMSO-d6) δ 11.12 (s, 1H), 10.99 (s, 1H), 8.24 (t, J = 5.6 Hz, 1H), 7.98 (s, 1H), 7.58 (q, J = 8.6, 7.1 Hz, 1H), 7.47 (d, J = 8.3, 0.8 Hz, 1H), 7.43–7.38 (m, 2H), 7.25 (d, J = 7.6, 0.8 Hz, 1H), 7.17–7.01 (m, 4H), 6.82–6.77 (m, 1H), 6.74–6.66 (m, 1H), 6.61 (t, J = 5.8 Hz, 1H), 5.06 (dd, J = 12.9, 5.4 Hz, 1H), 4.83 (s, 2H), 3.82 (s, 3H), 3.61 (t, J = 5.4 Hz, 2H), 3.58–3.38 (m, 24H), 3.28–3.18 (m, 2H), 2.95–2.82 (m, 1H), 2.65–2.53 (m, 2H), 2.07–2.01 (m, 1H).

[0148] Example 40: 2-(5-Bromo-3-(4-(1-methyl-1H-indol-3-yl)-2,5-dioxo-2,5-dihydro-1H-pyrrol-3-yl)-1H-indol-1-yl)-N-(20-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-3,6,9,12,15,18-hexaoxa)acetamide (P7)

[0149]

[0150] The synthesis procedure of compound P7 is the same as that of P1, except that compound 12a is replaced with an equimolar amount of compound 12d, compound 8a is replaced with compound 8c, and triethylamine is replaced with an equimolar amount of N,N-diisopropylethylamine. The reaction gives a red solid P7 with a yield of 34.5%. 1HNMR(400MHz, DMSO-d6) δ 11.12 (s, 1H), 10.99 (s, 1H), 8.32 (t, J = 5.6 Hz, 1H), 7.83 (d, J = 2.7 Hz, 2H), 7.58 (t, J = 8.6, 7.1 Hz, 1H), 7.46 (d, J = 8.2 Hz, 1H), 7.30 (d, J = 8.7 Hz, 1H), 7.17–7.10 (m, 2H), 7.10–7.01 (m, 2H), 6.87 (d, J = 1.9 Hz, 1H), 6.77–6.66 (m, 2H), 6.61 (t, J = 5.8 Hz, 1H), 5.06 (dd, J = 12.9, 5.4 Hz, 1H), 4.91 (s, 2H), 3.88 (s, 3H), 3.61 (t, J = 5.4 Hz, 2H), 3.57–3.41 (m, 24H), 3.25 (q, J = 5.7 Hz, 2H), 2.95–2.83 (m, 1H), 2.63–2.52 (m, 2H), 2.07–2.01 (m, 1H).

[0151] Example 41: 2-(6-Bromo-3-(4-(1-methyl-1H-indol-3-yl)-2,5-dioxo-2,5-dihydro-1H-pyrrol-3-yl)-1H-indol-1-yl)-N-(20-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-3,6,9,12,15,18-hexaoxa)acetamide (P8)

[0152]

[0153] The synthesis procedure of compound P8 is the same as that of P1, except that compound 12a is replaced with an equimolar amount of compound 12d, compound 8a is replaced with compound 8d, and triethylamine is replaced with an equimolar amount of N,N-diisopropylethylamine. The reaction gives a red solid P8 with a yield of 37.0%. 1HNMR(400MHz, DMSO-d6) δ 11.12 (s, 1H), 10.99 (s, 1H), 8.33 (t, J = 5.6 Hz, 1H), 7.88 (s, 1H), 7.78 (s, 1H), 7.64–7.54 (m, 2H), 7.43 (d, J = 8.2 Hz, 1H), 7.14 (d, J = 8.6 Hz, 1H), 7.09–7.01 (m, 2H), 6.81–6.56 (m, 5H), 5.06 (dd, J = 12.9, 5.3 Hz, 1H), 4.93 (s, 2H), 3.87 (s, 3H), 3.61 (t, J = 5.4 Hz, 2H), 3.57–3.43 (m, 24H), 3.28 (q, J = 5.6 Hz, 2H), 2.95–2.81 (m, 1H), 2.65–2.52 (m, 2H), 2.07–2.00 (m, 1H).

[0154] Example 42: 2-(5-Methoxy-3-(4-(1-methyl-1H-indol-3-yl)-2,5-dioxo-2,5-dihydro-1H-pyrrol-3-yl)-1H-indol-1-yl)-N-(20-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-3,6,9,12,15,18-hexaoxa)acetamide (P9)

[0155]

[0156] The synthesis procedure of compound P9 was the same as that of P1, except that compound 12a was replaced with an equimolar amount of compound 12d, compound 8a was replaced with compound 8e, and triethylamine was replaced with an equimolar amount of N,N-diisopropylethylamine. The reaction yielded a red solid P9 with a yield of 51.7%. 1HNMR(400MHz,DMSO-d6)δ11.12(s,1H),10.94(s,1H),8.34(t,J=5.6Hz,1H),7.93(s,1H),7.72(s,1H),7.58(t,J=8.5,7.1Hz,1H),7.43(d,J=8.2Hz,1H),7.18(d,J=8.9Hz,1H),7.13(d,J=8.6Hz,1H),7.10–7.02(m,2H),6.92(d,J=8.0Hz,1H),6.71(t,J=7.5Hz,1H),6.61(t,J=5.9Hz,1H),6.56(dd,J=8.8,2.5Hz,1H),6.06(d,J=2.4Hz,1H),5.06(dd,J=13.0,5.3Hz,1H),4.90(s,2H),3.83(s,3H),3.61(t,J=5.4Hz,2H),3.57–3.42(m,J=5.8Hz,24H),3.27(q,J=5.6Hz,2H),2.95(s,3H),2.92–2.81(m,1H),2.66–2.52(m,2H),2.07–2.00(m,1H).

[0157] 2. Bioactivity test

[0158] Test method for enzyme level GSK-3β inhibitory activity: Express the C-terminal 6×His fusion GSK-3β protein through the Escherichia coli expression system and purify it by the Ni2+ affinity purification method. The kinase activity detection uses the Invitrongen Z-LYTE kinase kit with a 10L reaction system, and each sample has 3 replicates. The test samples are dissolved in DMSO and stored at low temperature (the concentration of DMSO in the final system is controlled within the range that does not affect the test activity). Use the enzyme-labeled Envision multi-label microplate detector (product of PerkinElmer) to detect the fluorescence intensities at 445 nM and 520 nM under 400 nM excitation, and calculate the substrate phosphorylation rate of the sample wells using the formula provided by the kit to reflect the kinase activity level. For those with an inhibition rate greater than 50%, the IC 50 value is obtained by fitting with GraphPad Prism software. The positive compound used in the experiment is Staurosporine. The GSK-3β inhibitory activities of the 9 synthesized target compounds P1 - P9 were tested, and the results are shown in the following table (Staurosporine is the control compound).

[0159]

[0160] The results of bioactivity tests showed that 9 compounds exhibited good inhibitory activity against GSK-3β, with IC 50 values ranging from 1.41 ± 0.29 to 5.46 ± 0.25 μM. Among them, compound P6 showed the best inhibitory activity (IC 50 = 1.41 ± 0.29 μM).

[0161] The content described in this specification is only a list of implementation forms of the inventive concept, and the protection scope of the present invention should not be regarded as limited to the specific forms stated in the embodiments.

Claims

1. A class of proteolysis-targeting chimeric compounds that target the degradation of GSK-3β, characterized in that This class of compounds has a general structural formula as shown in Formula P: ； Wherein: R is selected from 4-Br, and n = 6.

2. Use of a class of proteolysis-targeting chimera compounds targeting the degradation of GSK-3β as claimed in claim 1 for the preparation of a drug for regulating the GSK-3β signaling pathway.

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