A Artemisia ketonic acid amide derivative containing a nitrogen heterocycle, its preparation method and use

Abstract

The present invention relates to a nitrogen-containing heterocyclic derivative of rupestonic acid amide, its preparation method and use. The synthesis of this type of derivative is to first react rupestonic acid with nitrogen-containing heterocyclic compounds with different substitutions under the conditions of diisopropylamine and 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate to obtain 1-20 nitrogen-containing heterocyclic derivatives of rupestonic acid amide. The reaction conditions of this method are mild and the experimental steps are simple. The anti-tumor activities of the obtained 1-20 nitrogen-containing heterocyclic derivatives of rupestonic acid amide were tested on four types of cancer cells, namely Hela (human cervical cancer cells), HT-29 (human colon cancer cells), A549 (human non-small cell lung cancer cells) and HepG2 (human liver cancer cells). The experimental results show that compounds 1, 3, 14, 17, 18 and 19 are useful in the preparation of anti-tumor drugs.

Description

Technical Field

[0001] The present invention relates to a nitrogen-containing heterocyclic artemisia ketonic acid amide derivative, a preparation method thereof, and uses thereof. Background Art

[0002] Artemisia ketonic acid is a guaiane-type sesquiterpene compound isolated from the genuine medicinal material Artemisia rupestris. Research shows that guaiane-type sesquiterpene compounds have antitumor activity.

[0003] Nitrogen-containing heterocyclic compounds have a wide range of biological activities. Articles in international pharmaceutical chemistry journals often report that nitrogen-containing heterocyclic compounds have antitumor cell activity. Therefore, the present invention designed and synthesized a series of nitrogen-containing heterocyclic artemisia ketonic acid amide derivatives and conducted preliminary antitumor activity screening.

[0004] The present invention introduced nitrogen-containing heterocyclic compounds into the artemisia ketonic acid molecule to synthesize a series of nitrogen-containing heterocyclic artemisia ketonic acid amide derivatives, and the synthesis is rapid and simple. Summary of the Invention

[0005] The purpose of the present invention is to provide a nitrogen-containing heterocyclic artemisia ketonic acid amide derivative, a preparation method thereof, and uses thereof. The synthesis of this type of derivative is obtained by reacting artemisia ketonic acid with nitrogen-containing heterocyclic compounds with different substitutions under the conditions of diisopropylamine and 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate to obtain nitrogen-containing heterocyclic artemisia ketonic acid amide derivatives 1-20. This method has mild reaction conditions and simple experimental steps. Antitumor activity tests on four types of cancer cells, namely Hela (human cervical cancer cells), HT-29 (human colon cancer cells), A549 (human non-small cell lung cancer cells), and HepG2 (human liver cancer cells), were carried out on the obtained nitrogen-containing heterocyclic artemisia ketonic acid amide derivatives 1-20. The experimental results show that compounds 1, 3, 14, 17, 18, and 19 are useful in the preparation of antitumor drugs.

[0006] A nitrogen-containing heterocyclic artemisia ketonic acid amide derivative described in the present invention, and the structure of this derivative is as shown in general formula (I)

[0007]

[0008] Wherein: The R group is 4-(naphthalen-1-yl)piperazine, 4-(isoquinolin-1-yl)piperazine, 4-(naphthalen-1-ylmethyl)piperazine, piperidine, 4-methylpiperidine, 4-hydroxypiperidine, 4-(N,N-dimethylamino)piperidine, 4-(piperidin-1-yl)piperidine, 1,2,3,4-tetrahydroquinoline, 4-phenylpiperidine, 4-benzylpiperidine, 4-(4-fluorophenyl)piperidine, 4-(4-chlorophenyl)piperidine, 4-(4-bromophenyl)piperidine, 4-(4-cyanophenyl)piperidine, 4-(4-nitrophenyl)piperidine, 4-(4-trifluoromethoxyphenyl)piperidine, 4-(4-fluorobenzyl)piperidine, 4-(3-phenylpropyl)piperidine or indoline.

[0009] The preparation method of the abrotanine amide derivative containing a nitrogen heterocycle is carried out according to the following steps:

[0010] a. At room temperature, N,N-diisopropylethylamine, 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate and the nitrogen heterocyclic compound are successively added to the dichloromethane solution of abrotanine, and the reaction is carried out overnight. The progress of the reaction is monitored by TLC. After the reaction is complete, the reaction is quenched with water to obtain a mixture.

[0011] b. The mixture obtained in step a is extracted 3 times with 5 mL of ethyl acetate, and the extraction liquids are combined and washed successively with 5 mL of water and 10 mL of saturated brine, and then dried with anhydrous Na 2 SO 4 and the solvent is removed by rotary evaporation to obtain a crude product.

[0012] c. The crude product obtained in step b is purified by column chromatography using dichloromethane:methanol with an eluent volume ratio of 20:1 - 10:1 to obtain the target compounds 1 - 20.

[0013] The names of each compound are as follows:

[0014] Compound 1 is 4-(naphthalen-1-yl)piperazine-1-abrotanine amide;

[0015] Compound 2 is 4-(isoquinolin-1-yl)piperazine-1-abrotanine amide;

[0016] Compound 3 is 4-(naphthalen-1-ylmethyl)piperazine-1-abrotanine amide;

[0017] Compound 4 is piperidine-1-abrotanine amide;

[0018] Compound 5 is 4-methylpiperidine-1-abrotanine amide;

[0019] Compound 6 is 4-hydroxypiperidine-1-abrotanine amide;

[0020] Compound 7 is 4-(N,N-dimethylamino)piperidin-1-yl rupestonic acid amide;

[0021] Compound 8 is 4-(piperidin-1-yl)piperidin-1-yl rupestonic acid amide;

[0022] Compound 9 is 1,2,3,4-tetrahydroquinolin-1-yl rupestonic acid amide;

[0023] Compound 10 is 4-phenylpiperidin-1-yl rupestonic acid amide;

[0024] Compound 11 is 4-benzylpiperidin-1-yl rupestonic acid amide;

[0025] Compound 12 is 4-(4-fluorophenyl)piperidin-1-yl rupestonic acid amide;

[0026] Compound 13 is 4-(4-chlorophenyl)piperidin-1-yl rupestonic acid amide;

[0027] Compound 14 is 4-(4-bromophenyl)piperidin-1-yl rupestonic acid amide;

[0028] Compound 15 is 4-(4-cyanophenyl)piperidin-1-yl rupestonic acid amide;

[0029] Compound 16 is 4-(4-nitrophenyl)piperidin-1-yl rupestonic acid amide;

[0030] Compound 17 is 4-(4-trifluoromethoxyphenyl)piperidin-1-yl rupestonic acid amide;

[0031] Compound 18 is 4-(4-fluorobenzyl)piperidin-1-yl rupestonic acid amide;

[0032] Compound 19 is 4-(3-phenylpropyl)piperidin-1-yl rupestonic acid amide;

[0033] Compound 20 is indolin-1-yl rupestonic acid amide.

[0034] Use of compounds 1, 3, 14, 17, 18 and 19 in the a rupestonic acid amide derivative containing a nitrogen heterocycle for preparing drugs against tumor Hela, HT-29, A549 and HepG2 cells.

[0035] Compound 17 and 18 in the a rupestonic acid amide derivative containing a nitrogen heterocycle have inhibitory activity against Hela; compounds 1, 3, 14, 17, 18 and 19 have inhibitory activity against HT-29; compound 17 has inhibitory activity against A549; compounds 17, 18 and 19 have inhibitory activity against HepG2.

[0036] A kind of artemisitone amide derivative containing nitrogen heterocycle, its preparation method and use according to the present invention introduce a substituted nitrogen heterocycle into the artemisitone molecule to synthesize a series of artemisitone amide derivatives containing nitrogen heterocycle, and the synthesis route is as follows:

[0037]

[0038] Wherein: the R group is 4-(naphthalen-1-yl)piperazine, 4-(isoquinolin-1-yl)piperazine, 4-(naphthalen-1-ylmethyl)piperazine, piperidine, 4-methylpiperidine, 4-hydroxypiperidine, 4-(N,N-dimethylamino)piperidine, 4-(piperidin-1-yl)piperidine, 1,2,3,4-tetrahydroquinoline, 4-phenylpiperidine, 4-benzylpiperidine, 4-(4-fluorophenyl)piperidine, 4-(4-chlorophenyl)piperidine, 4-(4-bromophenyl)piperidine, 4-(4-cyanophenyl)piperidine, 4-(4-nitrophenyl)piperidine, 4-(4-trifluoromethoxyphenyl)piperidine, 4-(4-fluorobenzyl)piperidine, 4-(3-phenylpropyl)piperidine, indoline.

[0039] A kind of artemisitone amide derivative containing nitrogen heterocycle, its preparation method and use according to the present invention have carried out preliminary in vitro anti-tumor activity tests on the synthesized derivatives 1-20 against Hela, HT-29, A549 and HepG2 cells. The experimental results show that: some compounds show good activity, among which compounds 17 and 18 have good inhibitory effects on Hela (human cervical cancer cells) (IC 50 are 9.54 and 7.95 μmol / L respectively); compounds 1, 3, 14, 17, 18 and 19 have good inhibitory effects on HT-29 (human colon cancer cells) (IC 50 are 6.26, 9.11, 9.06, 4.90, 2.49 and 4.37 μmol / L respectively); compound 17 has a good inhibitory effect on A549 (human non-small cell lung cancer cells) (IC 50 is 10.31 μmol / L); compounds 17, 18 and 19 have good inhibitory effects on HepG2 (human liver cancer cells) (IC 50 are 5.44, 10.03 and 9.75 μmol / L respectively). Specific embodiments

[0040] The following examples are used to illustrate the present invention and do not constitute any limitation to the scope of the present invention.

[0041] Reagents:

[0042] Artemisitone is separated by a conventional method, purity: 98%, detected by HPLC, and the rest of the reagents are all commercially available analytical pure.

[0043] Example 1

[0044] Preparation of 4-(naphthalen-1-yl)piperazine-1-yl abrotanate amide (1):

[0045] a. At room temperature, 12 μL (0.07 mmol) of N,N-diisopropylethylamine, 25 mg (0.06 mmol) of 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, and 14 mg (0.06 mmol) of 4-(naphthalen-1-yl)piperazine were successively added to a 1 mL dichloromethane solution containing 14 mg (0.05 mmol) of abrotanate. The reaction was carried out overnight, and the progress of the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water to obtain a mixture;

[0046] b. The mixture obtained in step a was extracted 3 times with 5 mL of ethyl acetate, and the extraction liquids were combined and washed successively with 5 mL of water and 10 mL of saturated brine, and then dried over anhydrous Na 2 SO 4 and the solvent was removed by rotary evaporation to obtain a crude product;

[0047] c. The crude product obtained in step b was purified by column chromatography using dichloromethane:methanol with an eluent volume ratio of 100:1 to obtain the target compound 1, 4-(naphthalen-1-yl)piperazine-1-yl abrotanate amide, with a yield of 99%;

[0048] NMR data: 1 H NMR (500 MHz, CDCl 3 ) δ 8.21 (d, J = 7.8 Hz, 1H), 7.91–7.79 (m, 1H), 7.60 (d, J = 8.2 Hz, 1H), 7.54–7.44 (m, 2H), 7.41 (t, J = 7.8 Hz, 1H), 7.07 (d, J = 7.4 Hz, 1H), 5.35 (s, 1H), 5.17 (s, 1H), 3.87 (brs, 2H), 3.20–3.10 (m, 1H), 3.30–2.91 (m, 4H), 3.00 (d, J = 19.4 Hz, 1H), 2.82–2.77 (m, 1H), 2.63–2.52 (m, 2H), 2.33–2.17 (m, 2H), 2.18–2.08 (m, 1H), 2.05 (d, J = 18.7 Hz, 1H), 1.94–1.77 (m, 3H), 1.68 (s, 3H), 1.65–1.58 (m, 1H), 0.66 (d, J = 7.1 Hz, 3H); 13 C NMR (125 MHz, CDCl 3)δ208.0,173.8,170.7,148.8,148.7,137.9,134.7,128.7,128.5,126.0,125.7,125.7,124.2,123.0,115.0,113.5,45.9,41.3,40.6,38.6,37.7,36.6,35.2,30.4,30.3,12.0,8.1。

[0049] Example 2

[0050] Preparation of 4-(isoquinolin-1-yl)piperazine-1-yl abrotanate amide (2):

[0051] a. At room temperature, N,N-diisopropylethylamine (12 μL, 0.07 mmol), 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate (24 mg, 0.06 mmol) and 4-(naphthalen-1-yl)piperazine (13 mg, 0.06 mmol) were successively added to a CH 2 Cl 2 (1 mL) solution containing abrotanate (13 mg, 0.05 mmol). The reaction was carried out overnight, and the progress of the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water to obtain a mixture;

[0052] b. The mixture obtained in step a was extracted 3 times with 5 mL of ethyl acetate, and the extraction solutions were combined and washed successively with 5 mL of water and 10 mL of saturated brine, and then dried over anhydrous Na 2 SO 4 and the solvent was removed by rotary evaporation to obtain a crude product;

[0053] c. The crude product obtained in step b was purified by column chromatography using dichloromethane:methanol with an eluent volume ratio of 100:1 to obtain the target compound 2, 4-(isoquinolin-1-yl)piperazine-1-yl abrotanate amide, with a yield of 99%;

[0054] NMR data: 1 H NMR(500MHz,CDCl 3)δ8.14(d, J = 5.8 Hz, 1H), 8.10(d, J = 8.4 Hz, 1H), 7.78(d, J = 8.1 Hz, 1H), 7.64(ddd, J = 8.2, 6.8, 1.2 Hz, 1H), 7.55(ddd, J = 8.2, 6.8, 1.2 Hz, 1H), 7.31(d, J = 5.8 Hz, 1H), 5.35(s, 1H), 5.16(s, 1H), 4.05–3.80(m, 4H), 3.41(t, J = 5.0 Hz, 4H), 3.18–3.07(m, 1H), 2.99(d, J = 19.4 Hz, 1H), 2.83–2.70(m, 1H), 2.62–2.52(m, 2H), 2.17–2.07(m, 1H), 2.03(d, J = 18.8, 1H), 1.94–1.73(m, 3H), 1.67(s, 3H), 1.65–1.56(m, 1H), 0.64(d, J = 7.1 Hz, 3H); 13 C NMR(125 MHz, CDCl 3 )δ208.0, 173.9, 170.7, 160.7, 148.8, 140.4, 138.1, 137.9, 129.9, 127.3, 126.5, 125.0, 121.7, 116.7, 113.5, 47.4, 45.8, 41.3, 40.6, 38.5, 37.6, 36.5, 35.2, 30.4, 12.0, 8.0.

[0055] Example 3

[0056] Preparation of 4-(naphthalen-1-ylmethyl)piperazine-1-yl abrotanate amide (3):

[0057] a. At room temperature, 15 μL (0.09 mmol) of N,N-diisopropylethylamine, 30 mg (0.08 mmol) of 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, and 18 mg (0.08 mmol) of 4-(naphthalen-1-ylmethyl)piperazine were successively added to a 1 mL dichloromethane solution containing 17 mg (0.06 mmol) of abrotanate. The reaction was carried out overnight. The progress of the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water to obtain a mixture;

[0058] b. The mixture obtained in step a was extracted 3 times with 5 mL of ethyl acetate. The extracts were combined, washed successively with 5 mL of water and 10 mL of saturated brine, and then dried over anhydrous Na 2 SO 4 and the solvent was removed by rotary evaporation to obtain the crude product;

[0059] c. Purify the crude product obtained in step b by column chromatography using dichloromethane:methanol with an eluent volume ratio of 100:1 to obtain the target compound 3,4-(naphthalen-1-ylmethyl)piperazine-1-yl artemisia ketone amide with a yield of 99%;

[0060] NMR data: 1 H NMR(500MHz,CD 3 OD)δ8.30(dd,J=8.5,1.2Hz,1H),7.88–7.83(m,1H),7.81(d,J=7.5Hz,1H),7.54–7.45(m,2H),7.46–7.38(m,2H),5.37(d,J=1.2Hz,1H),5.12(s,1H),3.98(s,2H),3.74–3.49(m,4H),3.21–3.15(m,1H),2.93(d,J=19.4Hz,1H),2.78–2.67(m,1H),2.63–2.47(m,5H),2.11(td,J=6.3,3.4Hz,1H),2.06–1.97(m,2H),1.90–1.73(m,3H),1.63(d,J=1.6Hz,3H),1.63–1.55(m,1H),0.63(d,J=7.2Hz,3H); 13 CNMR(125MHz,CD 3 OD)δ210.9,177.9,172.6,150.0,138.5,135.4,134.2,133.8,129.5,129.5,129.1,126.9,126.8,126.1,125.8,114.8,61.6,47.2,42.3,41.8,38.9,38.7,37.6,36.6,31.5,30.8,12.2,8.0.

[0061] Example 4

[0062] Preparation of piperidin-1-yl artemisia ketone amide (4):

[0063] a. At room temperature, sequentially add 17 μL (0.10 mmol) of N,N-diisopropylethylamine, 19 mg (0.05 mmol) of 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, and 6 mg (0.05 mmol) of piperidine hydrochloride to a 1 mL dichloromethane solution containing 10 mg (0.04 mmol) of artemisia ketone acid. React overnight and monitor the reaction progress by TLC. After the reaction is complete, quench the reaction with water to obtain a mixture;

[0064] b. Extract the mixture obtained in step a three times with 5 mL of ethyl acetate. Combine the extracts, wash them successively with 5 mL of water and 10 mL of saturated brine, and then dry over anhydrous Na 2 SO 4 . Rotavapor to remove the solvent to obtain the crude product;

[0065] c. Purify the crude product obtained in step b by column chromatography using dichloromethane:methanol with an eluent volume ratio of 100:1 to obtain the target compound 4, piperidin-1-yl abrotanate amide, with a yield of 99%;

[0066] NMR data: 1 H NMR(500 MHz, CDCl 3 ) δ 5.24 (d, J = 1.2 Hz, 1H), 5.06 (s, 1H), 3.71–3.43 (m, 4H), 3.16–3.05 (m, 1H), 2.94 (d, J = 19.6 Hz, 1H), 2.72 (t, J = 11.5 Hz, 1H), 2.61–2.45 (m, 2H), 2.11 (tq, J = 7.0, 3.5 Hz, 1H), 2.02 (d, J = 18.8 Hz, 1H), 1.91–1.81 (m, 2H), 1.76 (tt, J = 14.1, 4.0 Hz, 1H), 1.71–1.63 (m, 5H), 1.62–1.50 (m, 5H), 0.63 (d, J = 7.1 Hz, 3H); 13 C NMR(125 MHz, CDCl 3 ) δ 208.1, 174.2, 170.5, 149.4, 137.8, 112.5, 45.9, 41.4, 40.6, 37.7, 36.6, 35.3, 30.4, 26.7, 25.7, 24.6, 12.0, 8.0.

[0067] Example 5

[0068] Preparation of 4-methylpiperidin-1-yl abrotanate amide (5):

[0069] a. At room temperature, successively add 13 μL (0.07 mmol) of N,N-diisopropylethylamine, 26 mg (0.07 mmol) of 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, and 7 mg (0.07 mmol) of 4-methylpiperidine to a 1 mL dichloromethane solution containing 14 mg (0.06 mmol) of abrotanate. React overnight and monitor the reaction progress by TLC. After the reaction is complete, quench the reaction with water to obtain a mixture;

[0070] b. Extract the mixture obtained in step a three times with 5 mL of ethyl acetate. Combine the extracts and wash them successively with 5 mL of water and 10 mL of saturated brine. Then dry over anhydrous Na 2 SO 4 and rotary evaporate to remove the solvent to obtain the crude product;

[0071] c. Purify the crude product obtained in step b by column chromatography using dichloromethane:methanol with an eluent volume ratio of 100:1 to obtain the target compound 5,4-methylpiperidin-1-artemisia ketone amide with a yield of 86%;

[0072] NMR data: 1 H NMR(500MHz,CDCl 3 )δ(500MHz,CDCl 3 )δ5.23(d,J=1.2Hz,1H),5.05(s,1H),4.57(s,1H),4.02(s,1H),3.11(dt,J=4.4,2.0Hz,1H),3.08–2.87(m,2H),2.71(t,J=11.6Hz,1H),2.68–2.60(m,1H),2.57(ddd,J=18.8,6.6,1.3Hz,1H),2.53–2.43(m,1H),2.11(tq,J=7.1,3.4Hz,1H),2.02(d,J=18.7Hz,1H),1.91–1.51(m,10H),1.17–1.01(m,2H),0.96(d,J=6.4Hz,3H),0.63(d,J=7.1Hz,3H); 13 C NMR(125MHz,CDCl 3 )δ208.1,174.1,170.4,149.4,137.8,112.5,47.5,45.8,41.3,37.7,36.6,35.3,34.9,33.8,31.2,30.2,21.7,12.0,8.0。

[0073] Example 6

[0074] Preparation of 4-hydroxypiperidin-1-artemisia ketone amide (6):

[0075] a. At room temperature, 12 μL (0.07 mmol) of N,N - diisopropylethylamine, 24 mg (0.06 mmol) of 2-(7 - azabenzotriazol - 1 - yl)-N,N,N',N' - tetramethyluronium hexafluorophosphate, and 6 mg (0.06 mmol) of 4 - hydroxypiperidine were successively added to a 1 mL dichloromethane solution containing 13 mg (0.05 mmol) of rupestonic acid. The reaction was carried out overnight. The progress of the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water to obtain a mixture.

[0076] b. The mixture obtained in step a was extracted 3 times with 5 mL of ethyl acetate. The extracts were combined and washed successively with 5 mL of water and 10 mL of saturated brine, and then dried over anhydrous Na 2 SO 4 . The solvent was removed by rotary evaporation to obtain a crude product.

[0077] c. The crude product obtained in step b was purified by column chromatography using dichloromethane:methanol with an eluent volume ratio of 50:1 to obtain the target compound 6, 4 - hydroxypiperidin - 1 - rupestonic acid amide, with a yield of 99%.

[0078] NMR data: 1 H NMR (500 MHz, CDCl 3 ) δ 5.25 (s, 1H), 5.07 (s, 1H), 4.09 (s, 1H), 3.95 (tt, J = 7.9, 3.7 Hz, 1H), 3.87 (s, 1H), 3.35–3.21 (m, 2H), 3.10 (dq, J = 6.5, 1.9 Hz, 1H), 2.93 (d, J = 19.6 Hz, 1H), 2.70 (t, J = 11.5 Hz, 1H), 2.63–2.45 (m, 3H), 2.10 (qd, J = 7.2, 3.5 Hz, 1H), 2.02 (d, J = 18.8 Hz, 1H), 1.92–1.80 (m, 4H), 1.80–1.72 (m, 1H), 1.64 (s, 3H), 1.61–1.46 (m, 3H), 0.62 (d, J = 7.1 Hz, 3H); 13 C NMR (125 MHz, CDCl 3 ) δ 208.1, 174.1, 170.5, 149.1, 137.8, 112.8, 66.9, 45.8, 41.3, 40.9, 40.5, 37.7, 36.5, 35.2, 30.3, 29.6, 12.0, 8.0.

[0079] Example 7

[0080] Preparation of 4-(N,N - dimethylamino)piperidin - 1 - rupestonic acid amide (7):

[0081] a. At room temperature, 10 μL (0.06 mmol) of N,N-diisopropylethylamine, 20 mg (0.05 mmol) of 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, and 7 mg (0.05 mmol) of 4-(N,N-dimethylamino)piperidine were successively added to a 1 mL dichloromethane solution containing 11 mg (0.04 mmol) of rupestonic acid. The reaction was carried out overnight, and the progress of the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water to obtain a mixture.

[0082] b. The mixture obtained in step a was extracted 3 times with 5 mL of ethyl acetate. The extraction solutions were combined, washed successively with 5 mL of water and 10 mL of saturated brine, and then dried over anhydrous Na 2 SO 4 and the solvent was removed by rotary evaporation to obtain a crude product.

[0083] c. The crude product obtained in step b was purified by column chromatography using dichloromethane:methanol with an eluent volume ratio of 100:1 to obtain the target compound 7, 4-(N,N-dimethylamino)piperidin-1-yl rupestonate, with a yield of 96%.

[0084] NMR data: 1 H NMR (500 MHz, CD 3 OD) δ 5.43 (d, J = 1.2 Hz, 1H), 5.21 (s, 1H), 4.73 (s, 1H), 4.29 (s, 1H), 3.52 (tt, J = 11.8, 3.7 Hz, 1H), 3.28–3.18 (m, 2H), 2.98 (d, J = 19.3 Hz, 1H), 2.90 (s, 6H), 2.75 (t, J = 11.4 Hz, 1H), 2.71–2.55 (m, 2H), 2.23–2.10 (m, 3H), 2.04 (d, J = 18.9 Hz, 1H), 1.92–1.78 (m, 3H), 1.72–1.58 (m, 7H), 0.66 (d, J = 7.1 Hz, 3H); 13 C NMR (125 MHz, CD 3 OD) δ 211.0, 178.0, 172.8, 150.1, 138.6, 114.9, 64.6, 47.2, 47.1, 42.4, 41.8, 40.5, 38.8, 37.6, 36.6, 31.7, 30.7, 12.2, 7.9.

[0085] Example 8

[0086] Preparation of 4-(piperidin-1-yl)piperidin-1-yl rupestonate (8):

[0087] a. At room temperature, 12 μL (0.07 mmol) of N,N - diisopropylethylamine, 23 mg (0.06 mmol) of 2-(7 - azabenzotriazol - 1 - yl)-N,N,N',N' - tetramethyluronium hexafluorophosphate, and 10 mg (0.06 mmol) of 4-(piperidin - 1 - yl)piperidine were successively added to a 1 mL dichloromethane solution containing 13 mg (0.05 mmol) of rupestonic acid. The reaction was carried out overnight, and the progress of the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water to obtain a mixture.

[0088] b. The mixture obtained in step a was extracted 3 times with 5 mL of ethyl acetate. The extraction liquids were combined and washed successively with 5 mL of water and 10 mL of saturated brine, and then dried with anhydrous Na 2 SO 4 and the solvent was removed by rotary evaporation to obtain a crude product.

[0089] c. The crude product obtained in step b was purified by column chromatography using dichloromethane:methanol with an eluent volume ratio of 15:1 to obtain the target compound 8, 4-(piperidin - 1 - yl)piperidine - 1 - rupestonic acid amide, with a yield of 99%.

[0090] NMR data: 1 H NMR (500 MHz, CDCl 3 ) δ 5.29 (s, 1H), 5.09 (s, 1H), 4.77 (s, 1H), 4.22 (s, 1H), 3.39–2.77 (m, 8H), 2.67 (t, J = 11.4 Hz, 1H), 2.59–2.45 (m, 2H), 2.43–2.21 (m, 2H), 2.15–1.92 (m, 5H), 1.89–1.64 (m, 6H), 1.63 (s, 3H), 1.60–1.40 (m, 4H), 0.61 (d, J = 7.2 Hz, 3H); 13 C NMR (125 MHz, CDCl 3 ) δ 207.9, 173.7, 170.6, 148.6, 137.8, 113.7, 63.7, 53.8, 53.4, 45.8, 41.2, 40.4, 37.7, 36.4, 35.1, 30.5, 29.6, 22.9, 22.4, 11.9, 8.0.

[0091] Example 9

[0092] Preparation of 1,2,3,4 - tetrahydroquinoline - 1 - rupestonic acid amide (9):

[0093] a. At room temperature, 11 μL (0.06 mmol) of N,N-diisopropylethylamine, 21 mg (0.05 mmol) of 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, and 7 mg (0.05 mmol) of 1,2,3,4-tetrahydroquinoline were successively added to a 1 mL dichloromethane solution containing 11 mg (0.04 mmol) of rupestonic acid. The reaction was carried out overnight, and the progress of the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water to obtain a mixture;

[0094] b. The mixture obtained in step a was extracted 3 times with 5 mL of ethyl acetate, and the extracts were combined and washed successively with 5 mL of water and 10 mL of saturated brine, and then dried over anhydrous Na 2 SO 4 and the solvent was removed by rotary evaporation to obtain a crude product;

[0095] c. The crude product obtained in step b was purified by column chromatography using dichloromethane:methanol with an eluent volume ratio of 100:1 to obtain the target compound 9, 1,2,3,4-tetrahydroquinolin-1-yl rupestonate, with a yield of 30%;

[0096] NMR data: 1 H NMR (500 MHz, CD 3 OD) δ 7.30 (d, J = 7.2 Hz, 1H), 7.24–7.19 (m, 1H), 7.15–7.09 (m, 2H), 5.44 (d, J = 1.2 Hz, 1H), 5.41 (s, 1H), 3.87 (ddd, J = 12.9, 6.9, 6.0 Hz, 1H), 3.83–3.75 (m, 1H), 3.02–2.92 (m, 1H), 2.85 (d, J = 19.6 Hz, 1H), 2.79 (t, J = 6.7 Hz, 2H), 2.59–2.50 (m, 2H), 2.39 (t, J = 10.7 Hz, 1H), 2.09–1.96 (m, 4H), 1.84–1.75 (m, 2H), 1.66–1.52 (m, 5H), 0.62 (d, J = 7.2 Hz, 3H); 13 C NMR (125 MHz, CD 3 OD) δ 210.8, 177.8, 173.2, 152.8, 139.8, 138.5, 129.7, 127.1, 126.7, 125.7, 118.2, 118.2, 47.1, 42.3, 40.8, 39.0, 37.7, 36.6, 32.2, 27.6, 25.2, 20.3, 12.2, 7.9.

[0097] Example 10

[0098] Preparation of 4-phenylpiperidin-1-yl abrotanate amide (10):

[0099] a. At room temperature, 12 μL (0.07 mmol) of N,N-diisopropylethylamine, 24 mg (0.06 mmol) of 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, and 10 mg (0.06 mmol) of 4-phenylpiperidine were successively added to a 1 mL dichloromethane solution containing 13 mg (0.05 mmol) of abrotanate. The reaction was carried out overnight, and the progress of the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water to obtain a mixture;

[0100] b. The mixture obtained in step a was extracted 3 times with 5 mL of ethyl acetate. The extraction solutions were combined and washed successively with 5 mL of water and 10 mL of saturated brine, and then dried over anhydrous Na 2 SO 4 and the solvent was removed by rotary evaporation to obtain a crude product;

[0101] c. The crude product obtained in step b was purified by column chromatography using dichloromethane:methanol with an eluent volume ratio of 100:1 to obtain the target compound 10, 4-phenylpiperidin-1-yl abrotanate amide, with a yield of 99%;

[0102] NMR data: 1 H NMR (500 MHz, CD 3 OD) δ 7.32–7.27 (m, 2H), 7.26–7.22 (m, 2H), 7.21–7.16 (m, 1H), 5.40 (d, J = 1.2 Hz, 1H), 5.19 (s, 1H), 4.71 (d, J = 13.2 Hz, 1H), 4.23 (d, J = 13.0 Hz, 1H), 3.28–3.24 (m, 1H), 3.01 (d, J = 19.3 Hz, 1H), 2.86 (ddt, J = 12.1, 7.5, 3.7 Hz, 2H), 2.77 (t, J = 11.4 Hz, 1H), 2.70–2.56 (m, 2H), 2.18–2.10 (m, 1H), 2.05 (d, J = 18.9 Hz, 1H), 1.97–1.79 (m, 5H), 1.75–1.55 (m, 7H), 0.67 (d, J = 7.1 Hz, 3H); 13 C NMR (125 MHz, CD 3OD) δ 210.9, 178.0, 172.7, 150.6, 146.6, 138.6, 129.6, 127.8, 127.5, 114.2, 47.3, 43.8, 43.4, 42.4, 38.8, 37.7, 36.7, 35.4, 34.1, 30.8, 12.2, 7.9。

[0103] Example 11

[0104] Preparation of 4-benzylpiperidin-1-yl abrotanate amide (11):

[0105] a. At room temperature, 14 μL (0.08 mmol) of N,N-diisopropylethylamine, 28 mg (0.07 mmol) of 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, and 13 mg (0.07 mmol) of 4-benzylpiperidine were successively added to a 1 mL dichloromethane solution containing 15 mg (0.06 mmol) of abrotanate. The reaction was carried out overnight, and the progress of the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water to obtain a mixture;

[0106] b. The mixture obtained in step a was extracted 3 times with 5 mL of ethyl acetate. The extraction solutions were combined and washed successively with 5 mL of water and 10 mL of saturated brine, and then dried over anhydrous Na 2 SO 4 and the solvent was removed by rotary evaporation to obtain a crude product;

[0107] c. The crude product obtained in step b was purified by column chromatography using dichloromethane:methanol with an eluent volume ratio of 100:1 to obtain the target compound 11, 4-benzylpiperidin-1-yl abrotanate amide, with a yield of 92%;

[0108] NMR data: 1 H NMR (500 MHz, CDCl 3 ) δ 7.30–7.25 (m, 2H), 7.22–7.17 (m, 1H), 7.13 (d, J = 7.1 Hz, 2H), 5.24 (s, 1H), 5.05 (s, 1H), 4.60 (s, 1H), 4.19–3.92 (m, 1H), 3.18–3.03 (m, 1H), 3.03–2.82 (m, 2H), 2.71 (t, J = 11.4 Hz, 1H), 2.66–2.42 (m, 5H), 2.16–2.07 (m, 1H), 2.03 (d, J = 18.9 Hz, 1H), 1.88–1.67 (m, 6H), 1.65 (s, 3H), 1.61–1.54 (m, 1H), 1.23–1.06 (m, 2H), 0.63 (d, J = 7.1 Hz, 3H);13 C NMR (125 MHz, CDCl 3 ) δ 208.0, 174.0, 170.4, 149.3, 139.7, 137.8, 129.0, 128.3, 126.1, 112.6, 47.5, 45.8, 42.9, 41.3, 38.3, 37.7, 36.5, 35.2, 32.8, 31.8, 30.2, 12.0, 8.0。

[0109] Example 12

[0110] Preparation of 4-(4-fluorophenyl)piperidin-1-yl abrotanate (12):

[0111] a. At room temperature, 12 μL (0.07 mmol) of N,N-diisopropylethylamine, 24 mg (0.06 mmol) of 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, and 11 mg (0.06 mmol) of 4-(4-fluorophenyl)piperidine were successively added to a 1 mL dichloromethane solution containing 13 mg (0.05 mmol) of abrotanate. The reaction was carried out overnight, and the progress of the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water to obtain a mixture;

[0112] b. The mixture obtained in step a was extracted 3 times with 5 mL of ethyl acetate, and the extraction solutions were combined and washed successively with 5 mL of water and 10 mL of saturated brine, and then dried over anhydrous Na 2 SO 4 and the solvent was removed by rotary evaporation to obtain a crude product;

[0113] c. The crude product obtained in step b was purified by column chromatography using dichloromethane:methanol with an eluent volume ratio of 100:1 to obtain the target compound 12, 4-(4-fluorophenyl)piperidin-1-yl abrotanate, with a yield of 92%;

[0114] NMR data: 1 H NMR (500 MHz, CD 3OD) δ 7.26 (dd, J = 8.7, 5.5 Hz, 2H), 7.02 (t, J = 8.8 Hz, 2H), 5.40 (d, J = 1.2 Hz, 1H), 5.19 (s, 1H), 4.70 (d, J = 13.1 Hz, 1H), 4.29–4.13 (m, 1H), 3.27–3.23 (m, 1H), 3.00 (d, J = 19.6 Hz, 1H), 2.93–2.83 (m, 2H), 2.77 (t, J = 12.0 Hz, 1H), 2.68–2.57 (m, 2H), 2.21–2.09 (m, 1H), 2.04 (d, J = 18.9 Hz, 1H), 1.97–1.79 (m, 5H), 1.75–1.53 (m, 7H), 0.66 (d, J = 7.2 Hz, 3H); 13 C NMR (125 MHz, CD 3 OD) δ 210.9, 178.0, 172.7, 162.9 (d, J = 242.6 Hz), 150.6, 142.6 (d, J = 2.8 Hz), 138.6, 129.5 (d, J = 7.5 Hz), 116.1 (d, J = 21.0 Hz), 114.2, 47.2, 43.3, 43.0, 42.4, 38.8 (d, J = 6.5 Hz), 37.7, 36.6, 35.4, 34.2, 30.7, 12.2, 7.9.

[0115] Example 13

[0116] Preparation of 4-(4-chlorophenyl)piperidin-1-yl abrotanate amide (13):

[0117] a. At room temperature, 15 μL (0.08 mmol) of N,N-diisopropylethylamine, 29 mg (0.08 mmol) of 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, and 15 mg (0.08 mmol) of 4-(4-chlorophenyl)piperidine were successively added to a 1 mL dichloromethane solution containing 16 mg (0.06 mmol) of abrotanate. The reaction was carried out overnight, and the progress of the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water to obtain a mixture.

[0118] b. The mixture obtained in step a was extracted 3 times with 5 mL of ethyl acetate. The combined extracts were washed successively with 5 mL of water and 10 mL of saturated brine, and then dried over anhydrous Na 2 SO 4 and the solvent was removed by rotary evaporation to obtain the crude product.

[0119] c. Purify the crude product obtained in step b by column chromatography using dichloromethane:methanol with an eluent volume ratio of 100:1 to obtain the target compound 13,4-(4-chlorophenyl)piperidine-1-yl rupicolinamide, with a yield of 96%;

[0120] NMR data: 1 H NMR(500MHz,CDCl 3 )δ7.29(d,J = 8.5Hz,2H),7.13(d,J = 8.4Hz,2H),5.30(d,J = 1.3Hz,1H),5.13(s,1H),4.80(s,1H),4.22(s,1H),3.24–3.05(m,2H),2.98(d,J = 19.8Hz,1H),2.88–2.68(m,4H),2.65–2.49(m,2H),2.14(dq,J = 7.3,3.6Hz,1H),2.05(d,J = 18.8Hz,1H),1.98–1.84(m,4H),1.79(t,J = 13.6Hz,1H),1.67(s,3H),1.65–1.54(m,2H),0.65(d,J = 7.1Hz,3H); 13 C NMR(125MHz,CDCl 3 )δ208.0,173.9,170.6,149.2,143.3,137.8,132.2,128.7,128.0,112.9,45.8,42.1,41.3,40.6,38.5,37.7,36.5,35.2,32.7,30.4,12.0,8.0。

[0121] Example 14

[0122] Preparation of 4-(4-bromophenyl)piperidine-1-yl rupicolinamide (14):

[0123] a. At room temperature, sequentially add 12 μL (0.07 mmol) of N,N-diisopropylethylamine, 23 mg (0.06 mmol) of 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate, and 15 mg (0.06 mmol) of 4-(4-bromophenyl)piperidine to a 1 mL dichloromethane solution containing 13 mg (0.05 mmol) of rupicolinic acid. React overnight, monitor the reaction progress by TLC, and after the reaction is complete, quench the reaction with water to obtain a mixture;

[0124] b. Extract the mixture obtained in step a three times with 5 mL of ethyl acetate, combine the extracts, wash them successively with 5 mL of water and 10 mL of saturated brine, and then use anhydrous Na 2 SO4 Dry it, remove the solvent by rotary evaporation to obtain the crude product;

[0125] c. Purify the crude product obtained in step b by column chromatography using dichloromethane:methanol with an eluent volume ratio of 100:1 to obtain the target compound 14, 4-(4-bromophenyl)piperidine-1-artemisia ketonic acid amide, with a yield of 99%;

[0126] Nuclear magnetic data: 1 H NMR(500MHz,CDCl 3 )δ7.42(d,J=8.4Hz,2H),7.06(d,J=8.3Hz,2H),5.28(s,1H),5.11(s,1H),4.78(s,1H),4.20(s,1H),3.19–3.02(m,2H),2.96(d,J=19.9Hz,1H),2.78–2.69(m,3H),2.61–2.47(m,2H),2.16–2.06(m,1H),2.03(d,J=18.9Hz,1H),1.93–1.82(m,4H),1.82–1.74(m,1H),1.65(s,3H),1.64–1.47(m,3H),0.63(d,J=7.1Hz,3H); 13 C NMR(125MHz,CDCl 3 )δ208.1,174.0,170.6,149.1,143.8,137.8,131.6,128.4,120.2,112.9,50.7,45.9,42.2,41.3,38.5,37.7,36.5,35.2,32.7,30.4,12.0,8.0。

[0127] Example 15

[0128] Preparation of 4-(4-cyanophenyl)piperidine-1-artemisia ketonic acid amide (15):

[0129] a. At room temperature, sequentially add 12 μL, 0.07 mmol of N,N-diisopropylethylamine, 23 mg, 0.06 mmol of 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate, and 11 mg, 0.06 mmol of 4-(4-cyanophenyl)piperidine to a 1 mL dichloromethane solution containing 13 mg, 0.05 mmol of artemisia ketonic acid. React overnight, monitor the progress of the reaction by TLC. After the reaction is complete, quench the reaction with water to obtain a mixture;

[0130] b. Extract the mixture obtained in step a three times with 5 mL of ethyl acetate. Combine the extracts, wash them successively with 5 mL of water and 10 mL of saturated brine, and then dry over anhydrous Na 2 SO 4 . Rotavapor to remove the solvent to obtain the crude product;

[0131] c. Purify the crude product obtained in step b by column chromatography using dichloromethane:methanol with an eluent volume ratio of 100:1 to obtain the target compound 15, 4-(4-cyanophenyl)piperidine-1-artemisia ketone amide, with a yield of 99%;

[0132] NMR data: 1 H NMR(500 MHz, CDCl 3 ) δ 7.60 (d, J = 8.3 Hz, 2H), 7.30 (d, J = 8.3 Hz, 2H), 5.30 (d, J = 1.2 Hz, 1H), 5.12 (s, 1H), 4.80 (s, 1H), 4.23 (s, 1H), 3.21–3.06 (m, 2H), 2.95 (d, J = 19.7 Hz, 1H), 2.85 (tt, J = 12.1, 3.5 Hz, 1H), 2.77–2.69 (m, 2H), 2.63–2.46 (m, 2H), 2.17–2.07 (m, 1H), 2.06–1.82 (m, 6H), 1.77 (tt, J = 13.9, 3.9 Hz, 1H), 1.64 (s, 3H), 1.62–1.55 (m, 2H), 0.63 (d, J = 7.1 Hz, 3H); 13 C NMR(125 MHz, CDCl 3 ) δ 208.1, 174.0, 170.7, 150.2, 149.0, 137.8, 132.5, 127.5, 118.7, 113.1, 110.5, 45.8, 42.8, 41.3, 40.5, 38.5, 37.7, 36.5, 35.2, 30.4, 29.6, 12.0, 8.0.

[0133] Example 16

[0134] Preparation of 4-(4-nitrophenyl)piperidine-1-artemisia ketone amide (16):

[0135] a. At room temperature, 15 μL (0.09 mmol) of N,N-diisopropylethylamine, 31 mg (0.08 mmol) of 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, and 17 mg (0.08 mmol) of 4-(4-nitrophenyl)piperidine were successively added to a 1 mL dichloromethane solution containing 17 mg (0.07 mmol) of rupestonic acid. The reaction was carried out overnight, and the progress of the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water to obtain a mixture.

[0136] b. The mixture obtained in step a was extracted 3 times with 5 mL of ethyl acetate. The extraction solutions were combined, washed successively with 5 mL of water and 10 mL of saturated brine, and then dried over anhydrous Na 2 SO 4 and the solvent was removed by rotary evaporation to obtain a crude product.

[0137] c. The crude product obtained in step b was purified by column chromatography using a dichloromethane:methanol eluent with a volume ratio of 50:1 to obtain the target compound 16, 4-(4-nitrophenyl)piperidin-1-yl rupestonate, with a yield of 99%.

[0138] NMR data: 1 H NMR (500 MHz, CDCl 3 ) δ 8.16 (d, J = 8.7 Hz, 2H), 7.36 (d, J = 8.7 Hz, 2H), 5.30 (d, J = 1.2 Hz, 1H), 5.13 (s, 1H), 4.82 (s, 1H), 4.25 (s, 1H), 3.28–3.07 (m, 2H), 3.03–2.86 (m, 2H), 2.82–2.70 (m, 2H), 2.61–2.47 (m, 2H), 2.16–2.07 (m, 1H), 2.02 (d, J = 19.0 Hz, 1H), 1.95 (d, J = 12.9 Hz, 2H), 1.91–1.82 (m, 2H), 1.77 (tt, J = 14.0, 3.9 Hz, 1H), 1.72–1.54 (m, 6H), 0.63 (d, J = 7.1 Hz, 3H); 13 C NMR (125 MHz, CDCl 3 ) δ 207.9, 173.8, 170.6, 152.3, 149.1, 146.7, 137.8, 127.5, 123.9, 113.1, 45.8, 42.7, 41.3, 40.6, 38.5, 37.7, 36.5, 35.2, 32.4, 30.4, 12.0, 8.0.

[0139] Example 17

[0140] Preparation of 4-(4-Trifluoromethoxyphenyl)piperidin-1-yl abrotanate amide (17):

[0141] a. At room temperature, 26 μL (0.15 mmol) of N,N-diisopropylethylamine, 28 mg (0.07 mmol) of 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, and 20 mg (0.07 mmol) of 4-(4-trifluoromethoxyphenyl)piperidine hydrochloride were successively added to a 1 mL dichloromethane solution containing 15 mg (0.06 mmol) of abrotanate. The reaction was carried out overnight, and the progress of the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water to obtain a mixture;

[0142] b. The mixture obtained in step a was extracted 3 times with 5 mL of ethyl acetate. The extraction solutions were combined and washed successively with 5 mL of water and 10 mL of saturated brine, and then dried over anhydrous Na 2 SO 4 and the solvent was removed by rotary evaporation to obtain a crude product;

[0143] c. The crude product obtained in step b was purified by column chromatography using dichloromethane:methanol with an eluent volume ratio of 50:1 to obtain the target compound 17, 4-(4-trifluoromethoxyphenyl)piperidin-1-yl abrotanate amide, with a yield of 99%;

[0144] NMR data: 1 H NMR (500 MHz, CDCl 3 ) δ 7.21 (d, J = 8.7 Hz, 2H), 7.15 (d, J = 8.4 Hz, 2H), 5.29 (s, 1H), 5.12 (s, 1H), 4.80 (s, 1H), 4.40–4.05 (m, 1H), 3.20–3.07 (m, 2H), 2.97 (d, J = 19.7 Hz, 1H), 2.85–2.69 (m, 3H), 2.62–2.47 (m, 2H), 2.17–2.08 (m, 1H), 2.03 (d, J = 18.9 Hz, 1H), 1.99–1.82 (m, 4H), 1.78 (tt, J = 13.9, 3.9 Hz, 1H), 1.69–1.51 (m, 6H), 0.64 (d, J = 7.1 Hz, 3H); 13 C NMR (125 MHz, CDCl 3)δ208.0,173.9,170.6,149.2,147.8,143.6,137.8,127.9,121.1,120.4(q,J=256.7Hz),112.9,47.7,45.9,42.1,41.3,40.6,37.7,36.6,35.2,34.1,30.3,12.0,8.0。

[0145] Example 18

[0146] Preparation of 4-(4-fluorobenzyl)piperidin-1-yl abrotanate amide (18):

[0147] a. At room temperature, 22 μL (0.12 mmol) of N,N-diisopropylethylamine, 24 mg (0.06 mmol) of 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, and 15 mg (0.06 mmol) of 4-(4-fluorobenzyl)piperidine hydrochloride were successively added to a 1 mL dichloromethane solution containing 13 mg (0.05 mmol) of abrotanate. The reaction was carried out overnight, and the progress of the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water to obtain a mixture;

[0148] b. The mixture obtained in step a was extracted 3 times with 5 mL of ethyl acetate, and the extraction solutions were combined and washed successively with 5 mL of water and 10 mL of saturated brine, and then dried over anhydrous Na 2 SO 4 and the solvent was removed by rotary evaporation to obtain a crude product;

[0149] c. The crude product obtained in step b was purified by column chromatography using dichloromethane:methanol with an eluent volume ratio of 50:1 to obtain the target compound 18, 4-(4-fluorobenzyl)piperidin-1-yl abrotanate amide, with a yield of 99%;

[0150] NMR data: 1 H NMR(500MHz,CDCl 3) δ 7.08 (dd, J = 8.4, 5.6 Hz, 2H), 6.96 (t, J = 8.7 Hz, 2H), 5.24 (s, 1H), 5.05 (s, 1H), 4.60 (s, 1H), 4.05 (s, 1H), 3.11 (dq, J = 6.8, 2.0 Hz, 1H), 3.05–2.84 (m, 2H), 2.70 (t, J = 11.5 Hz, 1H), 2.65–2.43 (m, 5H), 2.11 (dd, J = 7.3, 3.4 Hz, 1H), 2.02 (d, J = 18.9 Hz, 1H), 1.90–1.68 (m, 6H), 1.64 (s, 3H), 1.62–1.51 (m, 1H), 1.24–0.97 (m, 2H), 0.63 (d, J = 7.1 Hz, 3H); 13 C NMR (125 MHz, CDCl 3 ) δ 208.0, 174.0, 170.4, 161.4 (d, J = 244.1 Hz), 149.3, 137.8, 135.3 (d, J = 3.5 Hz), 130.3 (d, J = 8.0 Hz), 115.1 (d, J = 20.9 Hz), 112.6, 47.4, 45.8, 42.0, 41.7, 41.3, 38.3, 37.7, 36.5, 35.2, 32.7, 31.7, 12.0, 8.0.

[0151] Example 19

[0152] Preparation of 4-(3-phenylpropyl)piperidine-1-yl abrotanate amide (19):

[0153] a. At room temperature, 13 μL (0.08 mmol) of N,N-diisopropylethylamine, 27 mg (0.07 mmol) of 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate, and 14 mg (0.07 mmol) of 4-(3-phenylpropyl)piperidine were successively added to a 1 mL dichloromethane solution containing 15 mg (0.06 mmol) of abrotanate. The reaction was carried out overnight, and the progress of the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water to obtain a mixture.

[0154] b. The mixture obtained in step a was extracted 3 times with 5 mL of ethyl acetate. The extraction solutions were combined, washed successively with 5 mL of water and 10 mL of saturated brine, and then dried over anhydrous Na 2 SO 4 and the solvent was removed by rotary evaporation to obtain the crude product.

[0155] c. Purify the crude product obtained in step b by column chromatography using dichloromethane:methanol with an eluent volume ratio of 50:1 to obtain the target compound 19, 4-(3-phenylpropyl)piperidine-1-yl rupicolinamide, with a yield of 98%;

[0156] NMR data: 1 H NMR (500 MHz, CDCl 3 ) δ 7.29–7.24 (m, 2H), 7.20–7.13 (m, 3H), 5.23 (s, 1H), 5.05 (s, 1H), 4.59 (s, 1H), 4.03 (s, 1H), 3.14–3.06 (m, 1H), 3.01–2.88 (m, 2H), 2.71 (t, J = 11.6 Hz, 1H), 2.66–2.45 (m, 5H), 2.11 (dq, J = 10.9, 5.3, 4.5 Hz, 1H), 2.02 (d, J = 18.8 Hz, 1H), 1.91–1.69 (m, 5H), 1.69–1.46 (m, 7H), 1.33–1.22 (m, 2H), 1.16–0.98 (m, 2H), 0.63 (d, J = 7.1 Hz, 3H); 13 C NMR (125 MHz, CDCl 3 ) δ 208.0, 174.1, 170.4, 149.3, 142.3, 137.8, 128.3, 128.2, 125.7, 112.5, 47.6, 45.8, 41.8, 41.3, 37.7, 36.5, 36.1, 36.0, 35.9, 35.2, 33.0, 31.9, 28.4, 12.0, 8.0.

[0157] Example 20

[0158] Preparation of indoline-1-yl rupicolinamide (20):

[0159] a. At room temperature, sequentially add 11 μL, 0.06 mmol of N,N-diisopropylethylamine, 22 mg, 0.06 mmol of 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, and 7 mg, 0.06 mmol of indoline to a 1 mL dichloromethane solution containing 12 mg, 0.05 mmol of rupicolinic acid. React overnight and monitor the progress of the reaction by TLC. After the reaction is complete, quench the reaction with water to obtain a mixture;

[0160] b. Extract the mixture obtained in step a three times with 5 mL of ethyl acetate. Combine the extracts, wash them sequentially with 5 mL of water and 10 mL of saturated brine, and then use anhydrous Na 2 SO 4Dry it, remove the solvent by rotary evaporation to obtain the crude product;

[0161] c. Purify the crude product obtained in step b by column chromatography using dichloromethane:methanol with an eluent volume ratio of 50:1 to obtain the target compound 20, indoline-1-artemisitone amide, with a yield of 93%;

[0162] NMR data: 1 H NMR(500MHz,CDCl 3 )δ8.08(s,1H),7.24–7.16(m,2H),7.05(t,J=7.4Hz,1H),5.43(d,J=1.2Hz,1H),5.34(s,1H),4.13(t,J=8.3Hz,2H),3.13(t,J=8.2Hz,3H),3.03(d,J=19.1Hz,1H),2.86(t,J=10.9Hz,1H),2.68–2.53(m,2H),2.19–2.08(m,1H),2.03(d,J=18.9Hz,1H),1.98–1.89(m,1H),1.89–1.80(m,1H),1.77(dt,J=13.9,3.4Hz,1H),1.74–1.62(m,4H),0.65(d,J=7.1Hz,3H); 13 C NMR(125MHz,CDCl 3 )δ208.0,173.8,169.4,150.7,142.3,137.9,132.4,127.4,124.9,124.2,117.1,114.4,53.4,45.8,41.3,40.2,37.9,36.6,35.3,30.8,29.7,12.0,8.0.

[0163] Example 21

[0164] Each artemisitone amide derivative containing a nitrogen heterocycle obtained in Examples 1-20 was subjected to a preliminary anti-tumor activity screening:

[0165] MTT method was used to detect cell viability:

[0166] Experimental procedure:

[0167] For the cells growing in the logarithmic growth phase, aspirate the culture medium, wash once with PBS, digest with trypsin, terminate with culture medium, gently pipette, count, and inoculate in a 96-well plate at the corresponding cell density (100 μL / well) and incubate overnight; add 20 μL / well of the sample solution, set a concentration gradient, and set 3 replicates for each concentration. CO 2Cultivate for 48 hours in an incubator at 37°C. Aspirate and discard the old culture medium, add 100 μL of MTT, and continue to cultivate for 2 hours. After incubating at 37°C for 2 hours, use an MB microplate reader to measure the light absorption value (OD) at 570 nm;

[0168] Calculation formula:

[0169] Percentage of cell viability % = (OD of compound - OD of blank / OD of control group - OD of blank) × 100%

[0170] Cell inhibition rate % = 1 - cell viability % = [1 - (OD of compound - OD of blank / OD of control group - OD of blank)] × 100%. Use graphpad, and the IC 50 ;

[0171] Sample treatment: The sample is 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 detection of activity;

[0172] Data processing and result description: Under the condition of a single concentration of 50 μM in the primary screening, the activity of the sample is tested; for the samples showing activity, see Table 1, the inhibition rate % (Inhibition) is greater than 75%, and further test the dose-dependent relationship of activity, that is, the IC 50 value, which is obtained by non-linear fitting of the sample concentration based on the sample activity. The software used for calculation is Graphpad Prism 4; each sample is set with duplicate wells (n ≥ 3) in the test and is represented by the standard deviation (Standard Deviation, SD) in the results. Each test has a reported compound as a reference. All data are made as reliable, accurate, and correct as possible within the scope of knowledge and ability;

[0173] Table 1 Inhibition rate results of tumor cells by nitrogeneous heterocyclic derivatives of artemisitone acid amide 1 - 20

[0174]

[0175] Note: The results in Table 1 are the inhibition rates of tumor cells measured under the condition of a monomer concentration of 50 μM;

[0176] The inhibition rate of the positive control DOX (doxorubicin) in Table 1 is the inhibition rate of tumor cells measured under the condition of 1 μM;

[0177] Table 2 Anti-tumor activity results of nitrogeneous heterocyclic derivatives of artemisitone acid amide 1 - 20

[0178]

[0179]

[0180] Note: IC 50 refers to the activity-dose dependence relationship of the sample measured when the inhibition rate is greater than 75% at a monomer concentration of 50 μM;

[0181] As can be seen from Table 2, the in vitro anti-tumor activities of the artemisitone acid amide derivatives 1-20 containing nitrogen heterocycles against Hela, HT-29, A549 and HepG2 cells were preliminarily tested; the experimental results showed that some compounds exhibited good activities, among which compounds 17 and 18 had good inhibitory effects on Hela (human cervical cancer cells) (IC 50 were 9.54 and 7.95 μmol / L, respectively); compounds 1, 3, 14, 17, 18 and 19 had good inhibitory effects on HT-29 (human colon cancer cells) (IC 50 were 6.26, 9.11, 9.06, 4.90, 2.49 and 4.37 μmol / L, respectively); compound 17 had good inhibitory effects on A549 (human non-small cell lung cancer cells) (IC 50 was 10.31 μmol / L); compounds 17, 18 and 19 had good inhibitory effects on HepG2 (human liver cancer cells) (IC 50 were 5.44, 10.03 and 9.75 μmol / L, respectively).

Claims

1. A derivative of artemisitone acid amide containing a nitrogen heterocycle, characterized in that the structure of this derivative is as shown in general formula (I) wherein: the R group is 4-(naphthalen-1-yl)piperazine, 4-(isoquinolin-1-yl)piperazine, 4-(naphthalen-1-ylmethyl)piperazine, piperidine, 4-methylpiperidine, 4-hydroxypiperidine, 4-(N,N-dimethylamino)piperidine, 4-(piperidin-1-yl)piperidine, 1,2,3,4-tetrahydroquinoline, 4-phenylpiperidine, 4-benzylpiperidine, 4-(4-fluorophenyl)piperidine, 4-(4-chlorophenyl)piperidine, 4-(4-bromophenyl)piperidine, 4-(4-cyanophenyl)piperidine, 4-(4-nitrophenyl)piperidine, 4-(4-trifluoromethoxyphenyl)piperidine, 4-(4-fluorobenzyl)piperidine, 4-(3-phenylpropyl)piperidine, indoline.

2. A preparation method of the derivative of artemisitone acid amide containing a nitrogen heterocycle according to claim 1, characterized in that it is carried out according to the following steps: a. At room temperature, N,N-diisopropylethylamine, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and the nitrogen heterocyclic compound are successively added to the dichloromethane solution containing artemisitone acid, and the reaction is carried out overnight. The progress of the reaction is monitored by TLC. After the reaction is complete, the reaction is quenched with water to obtain a mixture; b. Extract the mixture obtained in step a three times with 5 mL of ethyl acetate, combine the extracts, wash them successively with 5 mL of water and 10 mL of saturated brine, and then dry over anhydrous Na 2 SO 4 dry, and remove the solvent by rotary evaporation to obtain the crude product; c. The crude product obtained in step b is purified by column chromatography using dichloromethane:methanol with an eluent volume ratio of 20:1 to 10:1 to obtain the target compounds 1-20; The names of its various compounds are: Compound 1 is 4-(naphthalen-1-yl)piperazine-1-artemisitone acid amide; Compound 2 is 4-(isoquinolin-1-yl)piperazine-1-artemisitone acid amide; Compound 3 is 4-(naphthalen-1-ylmethyl)piperazine-1-artemisitone acid amide; Compound 4 is piperidine-1-artemisitone acid amide; Compound 5 is 4-methylpiperidine-1-artemisitone acid amide; Compound 6 is 4-hydroxypiperidine-1-artemisitone acid amide; Compound 7 is 4-(N,N-dimethylamino)piperidine-1-artemisitone acid amide; Compound 8 is 4-(piperidin-1-yl)piperidine-1-artemisitone acid amide; Compound 9 is 1,2,3,4-tetrahydroquinoline-1-artemisitone acid amide; Compound 10 is 4-phenylpiperidine-1-artemisitone acid amide; Compound 11 is 4-benzylpiperidine-1-artemisitone acid amide; Compound 12 is 4-(4-fluorophenyl)piperidine-1-artemisitone acid amide; Compound 13 is 4-(4-chlorophenyl)piperidine-1-artemisitone acid amide; Compound 14 is 4-(4-bromophenyl)piperidine-1-artemisitone acid amide; Compound 15 is 4-(4-cyanophenyl)piperidine-1-artemisitone acid amide; Compound 16 is 4-(4-nitrophenyl)piperidine-1-artemisitone acid amide; Compound 17 is 4-(4-trifluoromethoxyphenyl)piperidine-1-artemisitone acid amide; Compound 18 is 4-(4-fluorobenzyl)piperidine-1-artemisitone acid amide; Compound 19 is 4-(3-phenylpropyl)piperidin-1-yl rupestonic acid amide; Compound 20 is indolin-1-yl rupestonic acid amide.

3. Among the rupestonic acid amide derivatives containing nitrogen heterocycles obtained by the method according to claim 2: the use of Compounds 17 and 18 in the preparation of drugs for anti-tumor Hela cells; The use of Compounds 1, 3, 14, 17, 18 and 19 in the preparation of drugs for anti-tumor HT-29 cells; The use of Compound 17 in the preparation of drugs for anti-tumor A549 cells; The use of Compounds 17, 18 and 19 in the preparation of drugs for anti-tumor HepG2 cells.

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