Dihydroquinazolinone compound and preparation method thereof

Abstract

The invention provides a dihydroquinazolinone compound and a preparation method thereof. The compound is synthesized for the first time and has a brand new skeleton type. Meanwhile, a pharmacological test shows that the compound and the pharmaceutical composition thereof can have a remarkable inhibiting effect on cancers. Particularly, a pharmacological test shows that the pharmaceutical composition has a remarkable treatment effect on prostatic cancer, and the effect is obviously superior to that of an existing contrast drug.

Description

Technical Field

[0001] The invention belongs to the technical field of medicines, and specifically relates to a dihydroquinazolinone compound and a preparation method thereof. Background Art

[0002] Dihydroquinazolinone compounds are a class of organic molecules with unique structural features and diverse active functions. Their core feature is the dihydroquinazolinone skeleton they contain. Dihydroquinazolinone compounds are of great interest to researchers as organic compounds with extensive research and application value. Dihydroquinazolinone compounds have shown great application potential in the field of pesticides. They are currently used as highly efficient and low-toxic herbicides, which achieve the effect of weed control by interfering with the growth process of plants. Compared with traditional chemical herbicides, dihydroquinazolinone herbicides have higher environmental compatibility and biosafety, less pollution to soil and water bodies, and less impact on subsequent crops. Therefore, they have attracted much attention from experts in the agricultural field. Moreover, with the continuous discovery and progress of scientific researchers, this type of compound has shown a wide range of biological activities in the medical field, including pharmacological effects such as antibacterial and anticonvulsant, and is one of the important candidate structures in drug development. In addition, traditional synthesis methods require the use of toxic and harmful reagents and are carried out at high temperature, high pressure or in organic solvents, which pollutes the environment; and some synthesis methods have low yields and are difficult to meet the needs of large-scale production, limiting the commercial application of dihydroquinazolinone compounds. Although dihydroquinazolinone compounds have a wide range of biological activity characteristics, there are still some urgent problems to be solved in disease treatment or application environments, such as poor efficacy of existing tumor drugs, serious drug toxicity and side effects, and easy drug resistance, which are difficult to meet the needs of high-efficiency and low-toxicity treatment, and some disclosed dihydroquinazolinone compounds have deficiencies in stability, water solubility, etc. Summary of the invention

[0003] To solve the above technical problems, the present invention provides a dihydroquinazolinone compound and a preparation method thereof. In addition, pharmacological tests of the present invention show that the dihydroquinazolinone compound provided for the first time by the present invention has significant anticancer activity.

[0004] The present invention provides the following compounds, pharmaceutically acceptable salts, stereoisomers or deuterated derivatives thereof, selected from:

[0005]

[0006]

[0007] A pharmaceutical composition comprising any one of the above-mentioned compounds, its pharmaceutically acceptable salts, stereoisomers or deuterated compounds and a pharmaceutically acceptable carrier. The pharmacological tests of the present invention show that the dihydroquinazolinone compound provided for the first time in the present invention has significant anti-cancer activity. Therefore, the present invention provides that the above-mentioned compounds, their pharmaceutically acceptable salts, stereoisomers, deuterated compounds or the above-mentioned pharmaceutical compositions can be used for preventing or treating cancer.

[0008] Beneficial effects: A new dihydroquinazolinone compound is synthesized in the present invention. The compound of the present invention is synthesized for the first time and has a completely new skeleton type. At the same time, pharmacological tests show that the compound of the present invention and its pharmaceutical composition can have a significant inhibitory effect on cancer. In particular, pharmacological tests show that the present invention has a significant therapeutic effect on prostate cancer, and the effect is significantly better than that of the existing control drugs. Description of the Drawings

[0009] Figure 1 For the compound to inhibit the growth of prostate cancer 22RV1 and VCaP cells. Detailed Embodiments

[0010] The following further elaborates the present invention in conjunction with specific examples and test examples, but does not limit the scope of the present invention in any form.

[0011] Example 1: Synthesis of Compound 1

[0012]

[0013] Synthesis route:

[0014]

[0015] Step 1: Synthesis of Compound 1-2

[0016] Compound 1-1 (0.5 g, 2.051 mmol, 1 eq), 4-(Boc-amino)piperidine-4-carboxylic acid (0.314 g, 2.05 mmol, 1 eq) and sodium bicarbonate (0.808 g, 9.62 mmol, 4.7 eq) were added to a mixed solvent of 2 ml of acetonitrile and 6 ml of water. The reaction was carried out at 80 °C for 24 h under nitrogen protection. After monitoring the reaction to completion by thin layer chromatography, ethyl acetate was added to the reaction solution for extraction, and the organic phase was discarded. A certain amount of methanol was added to the aqueous phase, and the aqueous phase was concentrated by rotary evaporation. The pH was adjusted to 4.5 with 3 mol / L hydrochloric acid aqueous solution, and the mixture was stirred at room temperature overnight. A large amount of solid precipitated, and the white solid compound 1-2 (0.366 g, 1.01 mmol, yield 49.48%) was obtained by suction filtration. (ESI) m / z = 262.1 [M - Boc] + . 11H NMR (300 MHz, DMSO-d 6 ) δ 12.44 (s, 1H), 11.71 (s, 1H), 8.13 (s, 1H), 7.35 (s, 1H), 7.17 (t, J = 2.9 Hz, 1H), 6.65–6.55 (m, 1H), 4.30 (d, J = 13.4 Hz, 2H), 3.57–3.24 (m, 4H), 2.10–1.77 (m, 4H), 1.38 (s, 9H).

[0017] Step 2: Synthesis of Compounds 1-4

[0018] Compound 1-3 (0.5 g, 2.69 mmol, 1 eq) was added to 5 ml of dichloromethane and dissolved completely. Then di-tert-butyl dicarbonate (0.705 g, 3.23 mmol, 1.2 eq) was added, and the reaction was carried out at room temperature for 2 h. After monitoring the completion of the reaction by thin-layer chromatography, the reaction was stopped. The reaction solution was evaporated to dryness, then water (100 mL) was added to the residue, and the mixture was extracted with ethyl acetate (80 mL × 2), washed with saturated brine (70 mL × 2), dried over anhydrous sodium sulfate, filtered, evaporated to dryness, and purified by silica gel column chromatography to obtain white solid Compound 1-4 (0.765 g, 2.68 mmol, yield 99.4%). (ESI) m / z = 186.1 [M - Boc] + . 1 1H NMR (300 MHz, DMSO-d 6 ) δ 7.43 (s, 1H), 7.40–7.34 (m, 2H), 7.28 (d, J = 8.5 Hz, 2H), 4.66–4.47 (m, 2H), 3.28 (tt, J = 10.9, 6.0 Hz, 2H), 1.35 (s, 9H).

[0019] Step 3: Synthesis of Compound 1-5

[0020] Triphenylphosphine (0.367 g, 2 mmol, 1 eq) and iodine (0.355 g, 1.4 mmol, 2 eq) were dissolved in 3 ml of dichloromethane. After stirring at room temperature for 10 minutes, imidazole (0.19 g, 2.8 mmol, 4 eq) was added to the above reaction solution. After stirring at room temperature for 30 min, intermediate Compound 1-4 (0.2 g, 0.7 mmol, 1 eq) was added, and the reaction was stirred at room temperature for 2 h. After monitoring the completion of the reaction, 10% sodium sulfite and saturated sodium bicarbonate aqueous solution were added to the reaction solution. The mixture was extracted with dichloromethane (10 ml × 2), washed with saturated brine (10 mL × 2), evaporated to dryness, and purified by silica gel column chromatography to obtain white solid Compound 1-5 (0.268 g, 0.68 mmol, yield 97.14%). (ESI) m / z = 296.1 [M - Boc] + .

[0021] Step 4: Synthesis of Compound 1-6

[0022] Dissolve Compound 1-5 (0.39 g, 0.98 mmol, 1 eq) in acetonitrile (5 mL), stir at room temperature, and then add potassium carbonate (0.54 g, 3.91 mmol, 4 eq) and benzyl-1-piperazine carbonate (0.43 g, 1.95 mmol, 2 eq) to the above reaction solution. After stirring at room temperature for 10 minutes, transfer it to an oil bath and heat to 80 °C for overnight reaction. After monitoring the completion of the reaction, cool the reaction solution to room temperature, rotary evaporate the reaction solution, and purify it by silica gel column chromatography to obtain Compound 1-6 as a yellow oil (0.44 g, 0.90 mmol, yield 91.4%). (ESI) m / z = 388.2 [M - Boc] + . 1 H NMR (300 MHz, DMSO-d 6 ) δ 7.48 (d, J = 8.5 Hz, 1H), 7.36 (s, 6H), 7.30 (d, J = 8.6 Hz, 3H), 5.07 (s, 2H), 4.52 (q, J = 7.8 Hz, 1H), 2.25 (dt, J = 24.1, 6.1 Hz, 7H), 1.76 (dh, J = 26.9, 7.0 Hz, 3H), 1.35 (s, 9H), 1.25 (d, J = 9.8 Hz, 2H).

[0023] Step 5: Synthesis of Compound 1-7

[0024] Add Compound 1-6 (0.055 g, 0.113 mmol, 1 eq), palladium chloride (0.002 g, 0.011 mmol, 0.1 eq), and triethylamine (0.034 g, 0.338 mmol, 3 eq) to a two-necked flask and stir at room temperature for ten minutes. Then, under nitrogen protection, slowly add 1.5 ml of dichloromethane to the above reaction flask, dissolve triethylsilane (0.039 g, 0.338 mmol, 3 eq) in 0.5 ml of dichloromethane and slowly add it to the two-necked flask. After reacting at room temperature for 40 minutes, the reaction is completed. Rotary evaporate the reaction solution and directly use it for the next step. (ESI) m / z = 354.2 [M - Boc] + .

[0025] Step 6: Synthesis of Compound 1-9

[0026] 1-8 (8 g, 40.56 mmol) was dissolved in 80 mL of dichloromethane, and the mixture was stirred for 15 min under an ice bath. N-Bromosuccinimide (7.58 g, 42.59 mmol) was slowly added to the reaction flask in portions, and the reaction was continued in the ice bath for 2 h. 40 mL of water was added to the reaction solution, and the mixture was extracted with dichloromethane. The organic layer was washed twice with saturated brine, collected, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by silica gel column chromatography to obtain a yellow solid compound 1-9 (9.83 g, yield 88%). (ESI) m / z = 276.1 [M+H] + . 1 H NMR (300 MHz, DMSO-d 6 ) δ 7.63–7.43 (m, 5H), 7.40 (dd, J = 8.7, 2.4 Hz, 1H), 7.31 (d, J = 2.4 Hz, 1H), 7.24 (s, 2H), 6.86 (d, J = 8.9 Hz, 1H).

[0027] Step 7: Synthesis of compound 1-10

[0028] Compound 1-9 (9.83 g, 35.60 mmol) and tert-butylsulfinamide (12.94 g, 106.80 mmol) were dissolved in 60 mL of tetrahydrofuran. After dissolution, tetraethyl titanate (24.36 g, 106.80 mmol) was added. After the addition was complete, the reaction was carried out at 70 °C for 48 h. The reaction solution was cooled to room temperature, 35 mL of water and 70 mL of ethyl acetate were added dropwise, and a large amount of solid precipitated. The reaction solution was filtered through diatomaceous earth to remove the solid. The filtrate was extracted with ethyl acetate, the organic layer was washed with saturated brine, collected, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by silica gel column chromatography to obtain a yellow solid compound 1-10 (12.33 g, yield 91%). (ESI) m / z = 379.2 [M+H] + .

[0029] Step 8: Synthesis of compound 1-11

[0030] Compound 1-10 (12.33 g, 32.51 mmol) was dissolved in a mixed solution of 49 mL of tetrahydrofuran and 1 mL of water, stirred at room temperature, and sodium borohydride (4.92 g, 130.02 mmol) was slowly added in portions. The reaction was carried out at room temperature for 3 h. 25 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a white solid compound 1-11 (12.14 g, yield 98%). (ESI) m / z = 381.1 [M+H] + . 1 H NMR (300 MHz, DMSO-d 6) δ 7.42–7.20 (m, 5H), 7.12–7.00 (m, 2H), 6.61 (d, J = 8.6 Hz, 1H), 5.96 (d, J = 5.9 Hz, 1H), 5.48 (d, J = 6.1 Hz, 1H), 5.24 (s, 2H), 1.14 (s, 9H).

[0031] Step 9: Synthesis of Compound 1-12

[0032] Intermediate 1-11 (12.14 g, 31.84 mmol) was dissolved in 50 mL of tetrahydrofuran and stirred at room temperature. Trichloromethyl carbonate (14.17 g, 47.75 mmol) was slowly added, and the reaction was carried out at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure. 20 mL of water was added dropwise to the reaction flask, and saturated sodium carbonate solution was added dropwise to adjust the pH of the reaction solution to 7. Ethyl acetate was added to the reaction solution for extraction. The aqueous layer was extracted twice with ethyl acetate. The ethyl acetate layers were combined, washed with saturated brine, the organic layer was collected and dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by silica gel column chromatography to obtain yellow solid Compound 1-12 (9.28 g, yield 96%). (ESI) m / z = 303.2 [M+H] + .

[0033] Step 10: Synthesis of Compound 1-13

[0034] Compound 1-12 (0.56 g, 1.85 mmol) and cesium carbonate (1.81 g, 5.54 mmol) were dissolved in acetonitrile solution and stirred under ice bath conditions. Dimethyl sulfate (0.28 g, 2.22 mmol) was dissolved in acetonitrile and slowly added dropwise to the reaction flask. After the addition was completed, the ice bath was removed and the reaction was stirred at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure. 10 mL of water was added dropwise to the reaction flask, and extracted with ethyl acetate. The organic layer was collected and dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by silica gel column chromatography to obtain white solid 1-13 (0.41 g, yield 70%). (ESI) m / z = 317.2 [M+H] + . 1 H NMR (300 MHz, DMSO-d 6 ) δ 7.47–7.43 (m, 4H), 7.29–7.25 (m, 2H), 7.15 (dd, J = 11.6, 3.4 Hz, 2H), 6.95 (d, J = 8.1 Hz, 1H), 5.77 (s, 1H), 2.97 (s, 3H).

[0035] Step 11: Synthesis of Compound 1-14

[0036] Compound 1-13 (0.50 g, 1.58 mmol), 3,5-dimethylisoxazole-4-boronic acid pinacol ester (0.51 g, 1.73 mmol), sodium carbonate (0.50 g, 4.73 mmol) and tetrakis(triphenylphosphine)palladium(0) (0.18 g, 0.16 mmol) were dissolved in a mixed solution of 3 mL of toluene, 1 mL of ethanol and 3 mL of water, and stirred at 80 °C for 18 hours under nitrogen protection. The reaction solution was filtered through diatomaceous earth, and the filtrate was extracted with ethyl acetate. The organic layer was collected and washed once with saturated sodium bicarbonate and saturated brine respectively. The organic layer was collected and dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by silica gel column chromatography to obtain white solid compound 1-14 (0.36 g, yield 70%). (ESI) m / z = 334.2 [M+H] + . 1 H NMR (300 MHz, DMSO-d 6 ) δ 7.53–7.48 (m, 4H), 7.31–7.26 (m, 2H), 7.17 (dd, J = 11.6, 3.4 Hz, 2H), 6.92 (d, J = 8.1 Hz, 1H), 5.73 (s, 1H), 2.92 (s, 3H), 2.47 (s, 3H), 2.27 (s, 3H).

[0037] Step 11: Synthesis of compound 1-15

[0038] Compound 1-14 (0.15 g, 0.45 mmol) was dissolved in 2 mL of DMF, stirred in an ice bath, and sodium hydride (0.016 g, 0.67 mmol) was slowly added portionwise. After 10 minutes, ethyl 3-bromopropionate I-f13 (0.16 g, 0.90 mmol) was added dropwise to the reaction solution. After the addition was complete, the ice bath was removed, and the mixture was stirred at room temperature for 2 hours. 10 mL of saturated ammonium chloride solution was added, and the mixture was extracted with ethyl acetate. The organic layer was collected and dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by silica gel column chromatography to obtain pale yellow solid compound 1-15 (0.17 g, yield 89%). (ESI) m / z = 420.3 [M+H] + .

[0039] Step 12: Synthesis of compound 1-16

[0040] Compound 1-15 (0.17 g, 0.39 mmol) and lithium hydroxide monohydrate (0.02 g, 0.78 mmol) were dissolved in a mixed solution of 2 mL of methanol and 1 mL of water and reacted at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure, the pH was adjusted to 3 with 1 mol / L hydrochloric acid solution, and extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate, filtered, and the filtrate was rotary evaporated to dryness under reduced pressure to obtain a pale yellow viscous liquid compound 1-16 (0.14 g, yield 88%). (ESI) m / z = 406.2 [M+H] + . 1 H NMR (300 MHz, CDCl 3 ) δ 11.92 (s, 1H), 7.37–7.27 (m, 4H), 7.11 (dd, J = 8.4, 1.8 Hz, 1H), 6.98–6.90 (m, 2H), 5.61 (s, 1H), 3.97–3.85 (m, 1H), 3.41 (s, 3H), 2.77 (dd, J = 15.3, 8.2 Hz, 1H), 2.55 (dd, J = 11.3, 6.0 Hz, 1H), 2.31 (s, 3H), 2.17 (s, 3H), 1.36–1.18 (m, 2H).

[0041] Step 13: Synthesis of Compound 1-17

[0042] Compound 5-9 (0.11 g, 0.271 mmol, 1 eq), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (0.078 g, 0.407 mmol, 1.5 eq), 1-hydroxy-7-azabenzotriazole (0.055 g, 0.407 mmol, 1.5 eq) and N-methylmorpholine (0.082 g, 0.814 mmol, 3 eq) were added to 2 ml of DMSO and stirred at room temperature for 20 minutes. Then, compound 1-7 (0.101 g, 0.285 mmol, 1.1 eq) was added to the above reaction solution and reacted at room temperature for 3 h. After monitoring the completion of the reaction, the reaction was stopped, and the reaction solution was quenched with water (10 mL), extracted with ethyl acetate (10 mL × 2), washed with saturated brine (10 mL × 2), dried over anhydrous sodium sulfate, filtered, and rotary evaporated to dryness. The residue was purified by silica gel column chromatography to obtain a pale yellow oily compound 1-17 (0.115 g, 0.16 mmol, yield 57.2%). (ESI) m / z = 641.2 [M-Boc] + . 1 H NMR (300 MHz, CDCl 3)δ 7.39–7.29 (m, 6H), 7.28–7.23 (m, 3H), 7.11 (dd, J = 8.3, 2.0 Hz, 1H), 7.00 (d, J = 1.9 Hz, 1H), 6.93 (dd, J = 8.4, 4.3 Hz, 1H), 6.02 (s, 1H), 5.76 (s, 1H), 5.32 (s, 1H), 4.77 (s, 1H), 3.88 (ddd, J = 12.7, 7.0, 4.6 Hz, 1H), 3.42 (d, J = 1.9 Hz, 3H), 2.86 (dt, J = 15.2, 7.5 Hz, 1H), 2.58–2.44 (m, 1H), 2.34 (d, J = 1.5 Hz, 8H), 2.20 (d, J = 1.4 Hz, 3H), 1.97 (s, 1H), 1.53–1.14 (m, 13H).

[0043] Step 14: Synthesis of Compound 1-18

[0044] Trifluoroacetic acid (2 mL) was added dropwise to a solution of Compound 1-17 (100 mg, 0.14 mmol, 1 eq) in dichloromethane (10 mL) at 0 °C. The reaction mixture was stirred at 20 °C for 1 h. The reaction was monitored by LCMS until the starting material completely disappeared and the main peak corresponded to the target product. The reaction mixture was directly concentrated under reduced pressure to obtain Compound 1-18 (0.1 g, crude product), which was directly used in the next step. (ESI) m / z = 641.1 [M+H] + .

[0045] Step 15: Synthesis of Compound 1-19

[0046] Compound 1-2 (0.068 g, 0.188 mmol, 1 eq), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (0.052 g, 0.269 mmol, 1.4 eq), 1-hydroxy-7-azabenzotriazole (0.037 g, 0.269 mmol), and N-methylmorpholine (0.064 g, 0.628 mmol, 3.3 eq) were added to a round-bottom flask. Then, 2 mL of DMSO was added to the above mixture to dissolve it. After stirring at room temperature for 15 min, Compound 1-18 was added, and the reaction was carried out at room temperature for 6 h. After monitoring the completion of the reaction, the reaction was stopped. The reaction mixture was quenched with water (10 mL), extracted with ethyl acetate (10 mL × 2), washed with saturated brine (10 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain a pale yellow solid, Compound 1-19 (0.117 g, yield 66.3%). (ESI) m / z = 884.2 [M-Boc] + .

[0047] Step 16: Synthesis of Compound 1

[0048] To a solution of Compound 1-19 (100 mg, 0.10 mmol, 1 eq) in dichloromethane (10 mL) was added dropwise trifluoroacetic acid (2 mL) at 0 °C. The reaction mixture was stirred at 20 °C for 1 h. The reaction was monitored by LCMS until the starting material completely disappeared and the main peak corresponded to the target product. The reaction mixture was concentrated under reduced pressure to obtain Compound 1. (ESI) m / z = 884.7 [M+H] + . 1 H NMR (300 MHz, Chloroform-d) δ 10.20 (s, 1H), 8.64 (dd, J = 17.9, 6.9 Hz, 1H), 8.25 (d, J = 2.4 Hz, 1H), 7.31 (d, J = 6.5 Hz, 7H), 7.24 (t, J = 6.5 Hz, 1H), 7.17 (d, J = 8.3 Hz, 1H), 7.12 (dt, J = 8.3, 1.8 Hz, 1H), 7.07 (d, J = 3.7 Hz, 1H), 7.00 (t, J = 2.1 Hz, 1H), 6.97–6.89 (m, 1H), 6.50 (d, J = 3.6 Hz, 1H), 5.77 (d, J = 2.3 Hz, 1H), 5.02 (d, J = 8.0 Hz, 1H), 4.57 (t, J = 13.6 Hz, 2H), 3.98–3.82 (m, 1H), 3.72–3.56 (m, 3H), 3.55–3.38 (m, 6H), 3.25 (s, 2H), 2.74 (d, J = 7.8 Hz, 1H), 2.49–1.95 (m, 18H), 1.59 (d, J = 15.2 Hz, 2H).

[0049] Referring to the synthetic method of Compound 1, Compounds 3, 11-13, 17-20 were synthesized.

[0050]

[0051]

[0052]

[0053] Example 2: Synthesis of Compound 2

[0054]

[0055] Synthetic route:

[0056]

[0057] Step 1: Synthesis of Compound 2-1

[0058] Compound 1-4 (0.6 g, 1.8 mmol, 1 eq) was dissolved in 2 mL of DMF and stirred in an ice bath. Sodium hydride (0.64 g, 2.7 mmol, 1.5 eq) was added slowly in portions. After stirring for 10 minutes, ethyl bromoacetate (0.60 g, 3.6 mmol, 2 eq) was added dropwise. The mixture was stirred at room temperature for 2 hours, 10 mL of saturated ammonium chloride solution was added, and the mixture was extracted with ethyl acetate. The organic layer was collected, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by silica gel column chromatography to obtain a pale yellow solid 2-1 (0.71 g, 1.69 mmol, yield 94%). (ESI) m / z = 420.3 [M+H] + 。

[0059] Step 2: Synthesis of compound 2-2

[0060] Compound 2-1 (0.71 g, 1.69 mmol, 1 eq) and lithium hydroxide monohydrate (0.08 g, 3.39 mmol, 2 eq) were dissolved in a mixed solution of 2 mL of methanol and 1 mL of water and reacted at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure, the pH was adjusted to 3 with 1 mol / L hydrochloric acid solution, and the mixture was allowed to stand overnight. The white solid compound 2-2 (0.63 g, 1.61 mmol, yield 96%) was obtained by filtration. (ESI) m / z = 392.2 [M+H] + 。 1 1H NMR (300 MHz, DMSO-d 6 ) δ 12.59 (s, 1H), 7.47–7.14 (m, 7H), 7.09 (d, J = 8.5 Hz, 1H), 5.74 (s, 1H), 3.48 (d, J = 17.4 Hz, 5H), 2.32 (d, J = 3.4 Hz, 3H), 2.15 (d, J = 3.4 Hz, 3H).

[0061] Step 3: Synthesis of compound 2

[0062] 1-Hydroxy-7-azabenzotriazole (0.052 g, 0.383 mmol, 1.5 eq), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (0.074 g, 0.383 mmol, 1.5 eq), N-methylmorpholine (0.09 g, 0.894 mmol, 3.5 eq) and compound 2-2 (0.1 g, 0.256 mmol, 1 eq) were added into a eggplant-shaped flask, dissolved in 3 ml of DMSO. After stirring at room temperature for 30 min, 4-(1-piperazinyl)-1H-pyrrolo[2,3-d]pyrimidine (0.052 g, 0.256 mmol, 1 eq) was added and the reaction was carried out at room temperature for 5 h. After monitoring the completion of the reaction, the reaction was stopped. The reaction solution was quenched with water (10 mL), and a white solid precipitated. After filtration and drying, compound 2 (0.112 g, 0.19 mmol, yield 76.2%) was obtained. (ESI) m / z = 577.3 [M+H] + 。 1 H NMR (300 MHz, DMSO-d 6 ) δ 11.75 (s, 1H), 8.16 (s, 1H), 7.42–7.32 (m, 4H), 7.29 (d, J = 1.8 Hz, 2H), 7.27 (q, J = 1.8 Hz, 1H), 7.21 (dd, J = 3.6, 2.4 Hz, 1H), 7.11–7.06 (m, 1H), 6.61 (dd, J = 3.7, 1.9 Hz, 1H), 5.69 (s, 1H), 4.87 (d, J = 16.6 Hz, 1H), 3.88 (d, J = 5.7 Hz, 4H), 3.68–3.51 (m, 5H), 2.33 (s, 3H), 2.17 (s, 3H).

[0063] Compounds 10, 16, and 21 were synthesized by referring to the synthetic method of compound 2.

[0064]

[0065]

[0066] Example 3: Synthesis of Compound 4

[0067]

[0068] Synthetic route:

[0069]

[0070] Step 1: Synthesis of Compound 4-2

[0071] 4-1 (0.4 g, 1.57 mmol, 1 eq), 1-methyl-1H-pyrazole-5-boronic acid (0.391 g, 1.88 mmol, 1.2 eq) and potassium carbonate (0.649 g, 4.7 mmol, 3 eq) were added to a three-necked flask, followed by the addition of 10 mL of dioxane and 1 mL of water as a mixed solvent for dissolution, and the mixture was stirred at room temperature for 5 minutes under nitrogen protection. Bis(tri-tert-butylphosphine)palladium (0.04 g, 0.078 mmol, 0.5 eq) was added to the above reaction solution. The above reaction solution was heated to 80 °C under nitrogen protection and reacted for 4 hours. After monitoring the completion of the reaction by thin-layer chromatography, the oil bath was removed, the reaction solution was cooled to room temperature, the insoluble substances were removed by filtration, the reaction solution was rotary evaporated, and purified by silica gel column chromatography to obtain white solid compound 4-2 (0.24 g, 0.94 mmol, yield 59.7%). (ESI) m / z = 257.1 [M+H] + 。 1 H NMR (400 MHz, Chloroform-d) δ 7.65 (s, 1H), 7.55 (d, J = 2.0 Hz, 1H), 6.39 (d, J = 2.0 Hz, 1H), 3.91 (s, 3H), 3.83 (s, 3H).

[0072] Step 2: Synthesis of compound 4-3

[0073] Compound 4-2 (0.24 g, 0.935 mmol, 1 eq) and N-chlorosuccinimide (0.15 g, 1.12 mmol, 1.2 eq) were added to a sealed tube and heated to 70 °C in an oil bath for 3 h. The oil bath was removed, the reaction solution was cooled to room temperature, the reaction solution was rotary evaporated, and the insoluble substances were removed by stirring with methyl tert-butyl ether. The filtrate was rotary evaporated, 2 mL of methanol, 2 mL of tetrahydrofuran and 1 mL of 6N aqueous sodium hydroxide solution were added to the residue, and the mixture was stirred at room temperature overnight. After monitoring the completion of the reaction by thin-layer chromatography, the reaction was stopped, the reaction solution was rotary evaporated, 3N HCl was added to the residue to adjust the pH to 3, extracted with dichloromethane (10 mL × 2), washed with saturated brine (10 mL × 2), dried over anhydrous sodium sulfate, and rotary evaporated to obtain white solid product compound 4-3 (0.196 g, 0.71 mmol, yield 72%). (ESI) m / z = 277.1 [M+H] + 。 1 H NMR (300 MHz, DMSO-d 6 ) δ 7.84 (s, 1H), 7.71 (s, 1H), 3.73 (s, 3H).

[0074] Step 3: Synthesis of compound 4-4

[0075] Dissolve compound 1-14 (0.05 g, 0.15 mmol, 1 eq) in 2 ml of N,N-dimethylformamide. Transfer the above reaction solution to an ice bath, slowly add 60% sodium hydride (0.009 g, 0.225 mmol, 1.5 eq), stir for 5 minutes, and then add N-Boc-3-aminopropyl bromide (0.071 g, 0.3 mmol, 2 eq) to the above reaction solution. After the addition is complete, remove the ice bath and stir at room temperature for 1.5 h. After monitoring the reaction by thin layer chromatography until completion, add saturated ammonium chloride solution (10 mL), extract with ethyl acetate (10 mL×2), wash with saturated brine (10 mL×2), dry over anhydrous sodium sulfate, and rotary evaporate to obtain a crude product. Purify by silica gel column chromatography to obtain a transparent oily product 4-4 (0.064 g, 0.13 mmol, yield 86.6%). (ESI) m / z = 491.3 [M+H] + . 1 H NMR (300 MHz, Chloroform-d) δ 7.38–7.24 (m, 6H), 7.14 (dd, J = 8.4, 2.0 Hz, 1H), 7.00–6.93 (m, 2H), 5.49 (s, 1H), 3.95 (dt, J = 14.2, 7.0 Hz, 1H), 3.43 (s, 3H), 3.27 (dd, J = 13.6, 7.0 Hz, 1H), 3.00 (dt, J = 15.0, 5.8 Hz, 2H), 2.35 (s, 3H), 2.21 (s, 3H), 1.70 (s, 2H), 1.44 (s, 9H).

[0076] Step 4: Synthesis of compound 4-5

[0077] Dissolve compound 4-4 (0.25 g, 0.52 mmol) in 2 ml of 4N HCl / EA solution, stir at room temperature for 1 hour, stop the reaction after monitoring the reaction to completion, rotary evaporate the reaction solution, add saturated sodium bicarbonate solution (10 mL) to the residue, extract with ethyl acetate (10 mL×2), wash with saturated brine (10 mL×2), dry over anhydrous sodium sulfate, and rotary evaporate to obtain 0.22 g of a crude product, which is used directly in the subsequent reaction without further purification. (ESI) m / z = 391.2 [M+H] + .

[0078] Step 5: Synthesis of compound 4

[0079] Compound 4-3 (0.064 g, 0.23 mmol, 1 eq), 1-hydroxy-7-azabenzotriazole (0.047 g, 0.346 mmol, 1.5 eq), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (0.066 g, 0.346 mmol, 1.5 eq), and N-methylmorpholine (0.07 g, 0.69 mmol, 3 eq) were added to a round-bottom flask and dissolved in 3 mL of DMSO. After stirring the above reaction solution at room temperature for 20 min, compound 4-5 (0.09 g, 0.23 mmol, 1 eq) was added, and the reaction was carried out at room temperature for 4 h. After monitoring the completion of the reaction, water (10 mL) was added to the reactants, and a white solid precipitated. Filtration under suction gave white solid compound 4 (0.075 g, 0.11 mmol, yield 50.1%). (ESI) m / z = 649.2 [M+H] + 。 1 H NMR (300 MHz, DMSO-d6) δ 8.69 (t, J = 5.7 Hz, 1H), 7.73 (s, 2H), 7.40–7.20 (m, 7H), 7.03 (d, J = 8.5 Hz, 1H), 5.74 (d, J = 5.6 Hz, 1H), 3.92–3.81 (m, 1H), 3.73 (s, 2H), 3.22 (dd, J = 13.1, 6.5 Hz, 1H), 2.85 (ddd, J = 25.2, 16.2, 7.3 Hz, 1H), 2.36 (s, 3H), 2.19 (s, 3H), 1.78 (dp, J = 21.0, 6.8 Hz, 2H).

[0080] With reference to the synthetic method of compound 4, compounds 7, 8, and 14 were synthesized.

[0081]

[0082] Example 4: Synthesis of compound 5

[0083]

[0084] Synthetic route:

[0085]

[0086] Step 1: Synthesis of compound 4

[0087] Compound 1-2 (0.093 g, 0.256 mmol, 1 eq), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (0.074 g, 0.384 mmol, 1.5 eq), 1-hydroxy-7-azabenzotriazole (0.052 g, 0.384 mmol, 0.2 eq) and N-methylmorpholine (0.078 g, 0.768 mmol, 3 eq) were added to 3 ml of DMSO. After stirring at room temperature for 15 minutes, compound 4-5 (0.1 g, 0.256 mmol, 1 eq) was added to the above reaction solution. Subsequently, the reaction was continued at room temperature for 4 h. After monitoring the completion of the reaction, water (10 mL) was added to the reactants, and a white solid precipitated. Filtration by suction gave white solid compound 5 (0.145 g, 0.23 mmol, yield 91%). (ESI) m / z = 634.2 [M+H] + 。 1 H NMR (300 MHz, Chloroform-d) δ 10.43 (s, 1H), 8.34 (s, 1H), 8.11 (t, J = 6.2 Hz, 1H), 7.41–7.24 (m, 7H), 7.19–7.05 (m, 2H), 7.01–6.90 (m, 2H), 6.52 (d, J = 3.7 Hz, 1H), 5.50 (s, 1H), 4.53 (d, J = 13.5 Hz, 2H), 3.94 (dt, J = 14.2, 7.1 Hz, 1H), 3.61 (td, J = 11.1, 3.4 Hz, 2H), 3.44 (s, 4H), 3.07 (ddd, J = 25.6, 13.8, 6.2 Hz, 2H), 2.64 (s, 1H), 2.34 (s, 4H), 2.29 (d, J = 4.3 Hz, 1H), 2.20 (s, 3H), 1.54 (t, J = 13.4 Hz, 2H), 1.29–1.25 (m, 1H), 0.96–0.85 (m, 1H).

[0088] Referring to the synthesis method of compound 5, compounds 9, 15, and 22 were synthesized.

[0089]

[0090]

[0091] Example 6: Synthesis of compound 6

[0092]

[0093] Synthesis route:

[0094]

[0095] Step 1: Synthesis of Compound 6

[0096] Compound 2-2 (0.060 g, 0.15 mmol, 1 eq), N-((S)-1-amino-3-(3-fluorophenyl)propan-2-yl)-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)thiophene-2-carboxamide (0.063 g, 0.15 mmol, 1 eq) and N-methylimidazole (0.038 g, 0.46 mmol, 3 eq) were added to acetonitrile (10 mL). Then N,N,N',N'-tetramethylchloroformamidinium hexafluorophosphate (0.065 g, 0.23 mmol, 1.5 eq) was added to the above reaction solution. After reacting at room temperature for 4 hours, the reaction was monitored to be complete by thin layer chromatography. The above reaction solution was concentrated to dryness, saturated ammonium chloride solution (10 mL) was added to the residue, and it was extracted with ethyl acetate (10 mL × 2), washed with saturated brine (10 mL × 2), dried over anhydrous sodium sulfate, and concentrated to dryness. The product was purified by silica gel column chromatography to obtain a white solid product 6 (0.076 g, 0.095 mmol, yield 63.3%). (ESI) m / z = 800.3 [M+H] + 。 1 1H NMR (300 MHz, Chloroform-d) δ 7.50 (d, J = 3.6 Hz, 1H), 7.32 (d, J = 4.6 Hz, 1H), 7.30–7.26 (m, 2H), 7.23 (dd, J = 6.7, 2.6 Hz, 3H), 7.18 (dd, J = 3.4, 2.2 Hz, 1H), 7.17–7.13 (m, 1H), 7.11–6.84 (m, 5H), 6.80–6.69 (m, 1H), 5.48 (d, J = 23.9 Hz, 1H), 4.18 (dd, J = 42.3, 16.2 Hz, 2H), 3.98–3.79 (m, 1H), 3.73 (d, J = 8.9 Hz, 3H), 3.43 (d, J = 4.2 Hz, 3H), 3.32–3.04 (m, 3H), 2.69–2.41 (m, 1H), 2.29 (d, J = 17.3 Hz, 3H), 2.14 (d, J = 18.1 Hz, 3H).

[0097] Referring to the synthesis method of Compound 6, Compounds 23-25 were synthesized.

[0098]

[0099] Pharmacological Tests

[0100] Test for Antagonistic Effect on Cell Proliferation

[0101] The pharmaceutical composition involved in the present invention has undergone the following strict and systematic pharmacological tests to comprehensively evaluate its activity and therapeutic effect. These test data provide a strong scientific basis for the further research and development and clinical application of the compounds described in the present invention.

[0102] Quickly transfer the cryopreserved 22RV1 or Vcap cells taken out from liquid nitrogen to a water bath at 37 °C and quickly shake to thaw. Transfer the cell suspension to a 15 mL centrifuge tube, centrifuge at 1500 rpm for 5 min, discard the supernatant, resuspend the cell pellet with RPMI-1640 complete medium (containing 10% fetal bovine serum and 1% penicillin / streptomycin) and transfer it to a cell culture dish containing 10 mL of medium, and culture it in a 37 °C, 5% CO 2 incubator. Passage the cells after they adhere to the wall.

[0103] Take cells in the logarithmic growth phase with good condition, digest them with trypsin cell digestive solution, resuspend them with a certain amount of RPMI-1640 complete medium into a single cell suspension, count using a cell counting plate, inoculate them into a 96-well white plate at a density of 800 cells / well, let it stand for 5 min, and then place it in a 37 °C, 5% CO 2 incubator and culture overnight. The next day, add compounds at different concentrations to act on the cells, and treat the control group with DMSO at the corresponding concentration. Dilute the stock solution with RPMI-1640 complete medium to a final concentration of 30.00 μM, 10.00 μM, 3.33 μM, 1.11 μM, 0.37 μM, 0.12 μM, 0.04 μM, 0.01 μM. Set 3 parallel wells for each group, add 100 μL of the dosing solution to each well, and place it in a 37 °C, 5% CO 2 incubator and culture for 72 h.

[0104] After the drug has acted for 72 h, perform CellTitle-Glo detection. Add 100 μL of CellTitle-Glo solution to each well in the control group and the dosing group, mix on an oscillator for 2 minutes to induce cell lysis. Incubate the 96-well plate at room temperature for 10 minutes to stabilize the fluorescence signal value. Use a microplate reader to detect the luminescence signal. Take the cell group treated with DMSO as the control group, and calculate the half inhibitory concentration (50% Inhibitory Concentration, IC 50 ) according to the median effect equation: Inhibition rate (%) = (L value of the control group - L value of the drug treatment group) / A value of the control group × 100%. Use the GraphPad non-linear fitting formula to calculate the IC 50 of the compound. The results are shown in Table 1.

[0105] Table 1 Results of the inhibitory effect of the compounds of the present invention on cell proliferation

[0106] Compound Number <![CDATA[22RV1 IC 50 (μM)]]> <![CDATA[Vcap IC 50 (μM)]]> Capivasertib 12.42 >50 Compound 1 3.91 3.96 Compound 2 1.12 0.72 Compound 3 11.91 4.37 Compound 9 1.18 0.99 Compound 11 0.89 0.33 Compound 12 4.78 0.86 Compound 13 5.34 2.02 Compound 16 1.07 1.96 Compound 17 0.49 0.49 Compound 18 0.58 0.88 Compound 19 1.99 2.99 Compound 22 13.3 4.21

[0107] Results: The compounds showed good anti - proliferative activities in 22RV1 and Vcap cells, and their activities were better than those of the existing control drug Capivasertib. Among them, the activity results of compounds 2, 9, and 16 are shown in the appendix Figure 1 . In particular, the anti - tumor activities of compounds 1, 2, 9, 11, 12, 13, 16, 17, 18, and 19 were significantly better than those of the prior art control drug Capivasertib, showing significant prospects for drug application. Therefore, the present invention provides that the above - mentioned compounds, their pharmaceutically acceptable salts, stereoisomers, deuterated compounds, or the above - mentioned pharmaceutical compositions can be used for preventing or treating cancer.

[0108] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above - mentioned embodiments. Within the scope of the technical concept of the present invention, various equivalent transformations can be made to the technical solutions of the present invention, and these equivalent transformations all fall within the protection scope of the present invention. Additionally, it should be noted that the various specific technical features described in the above - mentioned specific embodiments can be combined in any appropriate manner without contradiction. To avoid unnecessary repetition, the present invention does not separately describe various possible combination methods. Furthermore, any arbitrary combination can be made between different embodiments of the present invention as long as it does not violate the idea of the present invention, and it should also be regarded as the content disclosed by the present invention.

Claims

1. A dihydroquinazolinone compound, characterized in that The dihydroquinazolinone compound is selected from:

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