A triptolide acrylamide derivative, its preparation method and use

Abstract

The application discloses a triacrylamide derivative of tripterine, and A1 and A2 are independently selected from O and NH; L is selected from -(CH2) n 、 -((CH2)2O) m (CH2)2-; n is an integer from 2 to 6, and m is an integer from 1 to 6; R1 and R2 are independently selected from H; R3 and R4 are independently selected from C1-C4 alkyl, or R3 and R4 are connected to each other to form a 5-7 membered heterocyclic ring containing one oxygen atom or a 5-7 membered heterocyclic ring containing one nitrogen atom. Compared with tripterine, the triacrylamide derivative of tripterine has a significantly enhanced inhibitory effect on tumor cells. The application further discloses a use of the triacrylamide derivative of tripterine in preparation of an antitumor drug.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of medicinal chemistry and pharmacology, and relates to celastrol acrylamide derivatives, a preparation method thereof and use thereof in the preparation of anti-tumor drugs. BACKGROUND

[0002] In recent years, the incidence and mortality of cancer have shown a rapid upward trend, which seriously endangers the physical and mental health of human beings. Although the prevention, diagnosis and treatment of cancer have developed rapidly, it is still one of the main killers leading to death.

[0003] Celastrol (CEL) is a kind of pentacyclic triterpenoid compounds derived from the roots of Chinese medicine Tripterygium wilfordii. A large number of studies have found that celastrol has various biological activities, such as anti-tumor, anti-inflammatory, anti-obesity, and anti-neurodegenerative diseases. The anti-tumor activity of celastrol is a research hotspot, and its activity still needs to be improved.

[0004] SUMMARY

[0005] The purpose of the present application is to modify and transform the structure of celastrol as a lead compound by introducing a Michael addition acceptor such as acrylamide, thereby providing a series of celastrol derivatives. Pharmacological experiments show that the anti-tumor activity of the celastrol derivatives of the present application is significantly improved compared with celastrol.

[0006] The purpose of the present application is achieved by the following technical solutions:

[0007] The celastrol acrylamide derivative has the structure shown in formula III:

[0008]

[0009] A1 and A2 are independently selected from O and NH;

[0010] L is selected from -(CH2) n - and -((CH2)2O) m (CH2)2-, n is an integer from 2 to 6, and m is an integer from 1 to 6;

[0011] R1 and R2 are independently selected from H, R3 and R4 are independently selected from C1-C4 alkyl, or R3 and R4 are connected to each other to form a 5-7 membered heterocyclic ring containing one oxygen atom together with the oxygen atom, or R3 and R4 are connected to each other to form a 5-7 membered heterocyclic ring together with the nitrogen atom.

[0012] Preferably, A1 and A2 are independently selected from O;

[0013] L is selected from -(CH2) n n is an integer from 3 to 5;

[0014] R1is selected from H, R2is selected from R3, R4are independently selected from C1-C4 alkyl, or R3, R4are mutually connected to form together with an oxygen atom a 5- to 7-membered heterocyclic ring containing one oxygen atom, or R3, R4are mutually connected to form together with a nitrogen atom a 5- to 7-membered heterocyclic ring;

[0015] or A1, A2are independently selected from NH;

[0016] L is selected from -(CH2) n -, -((CH2)2O) m (CH2)2-, n is an integer from 2 to 6, m is an integer from 1 to 3;

[0017] R1, R2are independently selected from H.

[0018] More preferably, A1, A2are independently selected from O;

[0019] L is selected from -(CH2) n -, n is an integer from 3 to 5;

[0020] R1is selected from H, R2is selected from R3, R4are independently selected from CH3, CH2CH3, CH(CH3)2, or R3, R4are mutually connected to form together with an oxygen atom a 6-membered heterocyclic ring containing one oxygen atom, or R3, R4are mutually connected to form together with a nitrogen atom a 6-membered heterocyclic ring;

[0021] or A1, A2are independently selected from NH;

[0022] L is selected from -(CH2) n -, -(CH2)2O(CH2)2-, -(CH2)2O(CH2)2O(CH2)2-, n is an integer from 3 to 6;

[0023] R1, R2are independently selected from H.

[0024] but not including: A1= A2= O, L is selected from -(CH2)5-, R1is selected from H, R2is selected from R3= R4= CH3, R3= R4= CH(CH3)2, or R3, R4are mutually connected to form together with a nitrogen atom a 6-membered heterocyclic ring.

[0025] Further more preferably, when A1, A2are independently selected from O, L is selected from -(CH2) n -, n is 3, R1is selected from H, R2is selected from R3, R4are independently selected from CH3; L is selected from -(CH2) n -; n is an integer from 3 to 5, R1is selected from H, and R2is selected from R3, R4are independently selected from CH2CH3; L is selected from -(CH2) n -; n is an integer from 3 to 5, R1is selected from H, and R2is selected from R3, R4are independently selected from CH2CH3; L is selected from -(CH2) n -; n is an integer from 3 to 5, R1is selected from H, and R2is selected from R3, R4are independently selected from CH2CH3; L is selected from -(CH2)

[0026] When A1, A2are independently selected from NH, L is selected from -(CH2) n -; n is an integer from 3 to 5, R1is selected from H, and R2is selected from

[0027] Specifically, the acrylamide derivative of tripterine is selected from the group consisting of:

[0028]

[0029]

[0030] As a further preferred technical solution of the present application, when A1, A2are independently selected from O, the acrylamide derivative of tripterine is selected from the group consisting of:

[0031]

[0032] Wherein, n, R3, R4are as described above.

[0033] Specifically, n is an integer from 2 to 6; R3, R4are independently selected from C1-C4 alkyl, or R3, R4are connected to each other to form a 5-7 membered heterocyclic ring containing one oxygen atom together with the oxygen atom, or R3, R4are connected to each other to form a 5-7 membered heterocyclic ring together with the nitrogen atom.

[0034] Preferably, n is an integer from 3 to 5; R3, R4are independently selected from C1-C4 alkyl, or R3, R4are connected to each other to form a 5-7 membered heterocyclic ring containing one oxygen atom together with the oxygen atom, or R3, R4are connected to each other to form a 5-7 membered heterocyclic ring together with the nitrogen atom.

[0035] More preferably, n is an integer from 3 to 5; R3, R4 are independently selected from CH3, CH2CH3, CH(CH3)2, or R3, R4 are connected to each other and to the oxygen atom to form a 6-membered heterocyclic ring containing one oxygen atom, or R3, R4 are connected to each other and to the nitrogen atom to form a 6-membered heterocyclic ring;

[0036] But not including: n is 5, R3 = R4 = CH3, R3 = R4 = CH(CH3)2, or R3, R4 are connected to each other and to the nitrogen atom to form a 6-membered heterocyclic ring.

[0037] More preferably, n is 3, R3, R4 are independently selected from CH3; n is an integer from 3 to 5, R3, R4 are independently selected from CH2CH3; n is an integer from 3 to 5, R3, R4 are connected to each other and to the oxygen atom to form a 6-membered heterocyclic ring containing one oxygen atom; n is an integer from 3 to 4, R3, R4 are connected to each other and to the nitrogen atom to form a 6-membered heterocyclic ring.

[0038] As a further preferred technical solution of the present application, when A1, A2 are independently selected from NH, the structure is shown in formula II:

[0039]

[0040] Wherein, L is as described above.

[0041] Specifically, L is selected from -(CH2) n -, -((CH2)2O) m (CH2)2-, n is an integer from 2 to 6, and m is an integer from 1 to 6. With the increase of L(-(CH2) n -, -((CH2)2O) m (CH2)2-), the inhibitory effect on tumor cells increases.

[0042] Preferably, L is selected from -(CH2) n -, -((CH2)2O) m (CH2)2-, n is an integer from 2 to 6, and m is an integer from 1 to 3.

[0043] More preferably, L is selected from -(CH2) n -, -(CH2)2O(CH2)2-, -(CH2)2O(CH2)2O(CH2)2-, n is an integer from 3 to 6.

[0044] More preferably, L is selected from -(CH2) n -, -(CH2)2O(CH2)2-, -(CH2)2O(CH2)2O(CH2)2-, n is an integer from 4 to 6.

[0045] Another object of the present application is to provide a preparation method of the emladin acrylamide derivative, when A1, A2 are independently selected from O, L is selected from -(CH2) n -, R1 is selected from H, and R2 is selected from as follows:

[0046]

[0047] wherein n, R3, R4 are as defined above;

[0048] comprising the following steps:

[0049] Step (1), reacting maleic anhydride with an amine compound represented by formula IV at room temperature to obtain intermediate V, using anhydrous dichloromethane as a reaction solvent and triethylamine as a base catalyst; wherein the molar ratio of maleic anhydride to the amine compound is 1:1.3-1:1.5; and the molar ratio of maleic anhydride to triethylamine is 1:1.3-1:1.5;

[0050] Step (2), reacting intermediate V with a dibromoalkane represented by formula at room temperature to obtain intermediate VI, using N,N-dimethylformamide (DMF) as a reaction solvent and K2CO3 as a base catalyst; wherein the molar ratio of intermediate V to the dibromoalkane is 1:3-1:4; and the molar ratio of intermediate V to K2CO3 is 1:2-1:3;

[0051] Step (3), reacting intermediate VI with emladin under heating reflux to obtain the emladin acrylamide derivative, using N,N-dimethylformamide (DMF) as a reaction solvent and NaHCO3 as a base catalyst; wherein the molar ratio of emladin to intermediate VI is 1:1.5-1:2; and the molar ratio of emladin to NaHCO3 is 1:3-1:4.

[0052] when A1, A2 are independently selected from NH, L is selected from -(CH2) n -, m (CH2)2-, R1, R2 are independently selected from H, as follows:

[0053]

[0054] wherein n, m, L are as defined above;

[0055] comprising the following steps:

[0056] Step (1), reacting maleic anhydride with an amine compound represented by formula IV at room temperature to obtain intermediate V, using anhydrous dichloromethane as a reaction solvent and triethylamine as a base catalyst; wherein the molar ratio of maleic anhydride to the amine compound is 1:1.3-1:1.5; and the molar ratio of maleic anhydride to triethylamine is 1:1.3-1:1.5; The N-(tert-butoxycarbonyl) diamine compound shown is reacted with a chloroacetamide shown in formula VII at room temperature to obtain intermediate VIII; wherein the molar ratio of the N-(tert-butoxycarbonyl) diamine compound to chloroacetamide is 1:1.5-1:2; and the molar ratio of the N-(tert-butoxycarbonyl) diamine compound to triethylamine is 1:2-1:3;

[0057] Step (2), intermediate VIII is subjected to acid hydrolysis to remove the Boc protecting group at room temperature in the presence of anhydrous dichloromethane as a reaction solvent and trifluoroacetic acid; wherein the volume ratio of trifluoroacetic acid to anhydrous dichloromethane is 1:5-1:10;

[0058] Step (3), amidation reaction occurs between intermediate IX and the carboxyl group of triptolide at room temperature in the presence of HATU and DIPEA with N,N-dimethylformamide (DMF) as a reaction solvent to obtain a triptolide acrylamide derivative; wherein the molar ratio of triptolide to intermediate IX is 1:4-1:5; the molar ratio of triptolide to HATU is 1:1.2-1:1.5; and the molar ratio of triptolide to DIPEA is 1:3-1:4.

[0059] Compared with triptolide, the triptolide acrylamide derivative of the present application has significantly enhanced inhibitory effect on tumor cells. Another object of the present application is to provide the use of the triptolide acrylamide derivative in the preparation of an anti-tumor drug.

[0060] The tumor is breast cancer, gastric cancer and colon cancer.

[0061] Another object of the present application is to provide a pharmaceutical composition which takes the triptolide acrylamide derivative as a main effective component and is supplemented with a pharmaceutically acceptable carrier to be prepared into any pharmaceutically acceptable dosage form.

[0062] The triptolide acrylamide derivative can be prepared into different dosage forms alone or in combination with one or more than one pharmaceutical carrier.

[0063] The dosage form is selected from tablets, capsules, dripping pills, granules, powders, lozenges, aqueous or oily suspensions, injections, patches, nano-preparations for oral, injection or topical administration in clinic. In these different preparations, the content of the triptolide acrylamide derivative of the present application can be 0.1%-99.9%.

[0064] The triptolide acrylamide derivative of the present application can be used alone or in combination with clinically commonly used anti-tumor drugs such as anti-metabolic drugs, alkylating agents, anti-tumor antibiotics, anti-tumor plant drugs, hormones and in combination with radiotherapy.

[0065] The beneficial effects of the present application are as follows:

[0066] The anti-tumor activity of the tripterine derivative of the present application is obviously improved compared with that of tripterine.

[0067] The preparation method of the tripterine derivative of the present application has mild reaction conditions, uses low-toxicity reagents, and has high yield. Specific embodiments

[0068] To further illustrate the technical solutions of the present application, a series of examples are listed below. These examples are illustrative and should not be construed as limiting the present application.

[0069] Example 1

[0070] Preparation of compound I-1

[0071]

[0072] Step (1), maleic anhydride (200 mg, 2.04 mmol) was dissolved in 5 mL of anhydrous dichloromethane, and dimethylamine (155 μL, 3.06 mmol) and triethylamine (425.3 μL, 3.06 mmol) were added in sequence. The reaction was stirred at room temperature, and the reaction progress was detected by TLC (dichloromethane:methanol = 20:1 V / V). After 1 h of reaction, it was detected that the raw material was completely reacted. After the reaction was completed, the organic phase was rotary evaporated to obtain the intermediate (E)-4-(dimethylamino)-4-oxobut-2-enoic acid, which was directly used in the next step reaction.

[0073]

[0074] Step (2), all the (E)-4-(dimethylamino)-4-oxobut-2-enoic acid obtained in step (1), K2CO3 (552.84 mg, 4 mmol) were stirred and dissolved in 4 mL of DMF, and 1,3-dibromopropane (609 μL, 6 mmol) was added. The reaction was carried out at room temperature, and the reaction progress was detected by TLC (petroleum ether: ethyl acetate = 1:2 V / V). After 4 h of reaction, it was detected that the reaction was completed. After the reaction was completed, 90 mL of ethyl acetate was added to dilute the reaction solution, and water washing (30 mL×3) was carried out. The organic phase was washed with saturated NaCl solution (100 ml×3), dried with anhydrous Na2SO4 overnight, and then filtered. The filtrate was concentrated, and then purified by normal phase silica gel column chromatography (eluent: petroleum ether: ethyl acetate = 3:2 V / V). After vacuum drying, compound A1 (white solid, 356.8 mg, yield about 66.2%) was obtained.

[0075]

[0076] Step (3), gossypol (90.12 mg, 0.2 mmol), compound A1 (105.65 mg, 0.4 mmol) and NaHCO3(67.2 mg, 0.8 mmol) were dissolved in 4 mL of DMF, and the reaction was heated to reflux at 60 °C. TLC was used to monitor the reaction progress (dichloromethane:methanol = 50:1 V / V). After 8 h, the reaction was determined to be complete. After the reaction was completed, 90 mL of ethyl acetate was added to dilute the reaction solution, which was washed with water (30 mL x 3). The organic phase was washed with saturated NaCl solution (100 mL x 3), dried over anhydrous Na2SO4overnight, filtered, and concentrated. The target compound I-1 (orange red powder, 83.2 mg, yield 65.7%) was obtained by normal phase silica gel column chromatography (eluent dichloromethane:methanol = 150:1 V / V) and vacuum drying.

[0077] m.p.: 229.4 °C.

[0078] ESI-MS: 634.3722 [M+H] + .

[0079] 1 H-NMR (300 MHz, DMSO-d6, TMS), δ ppm: 8.76 (s, 1H), 7.09 (d, J = 6.8 Hz, 1H), 6.70 (d, J = 11.9 Hz, 1H), 6.39 (s, 1H), 6.36 (d, J = 6.4 Hz, 1H), 5.95 (d, J = 12.1 Hz, 1H), 4.20-4.03 (m, 2H), 4.01-3.80 (m, 2H), 2.82 (s, 6H), 2.10 (s, 3H), 1.39 (s, 3H), 1.22 (s, 3H), 1.13 (s, 3H), 1.08 (s, 3H), 0.46 (s, 3H).

[0080] Example 2

[0081] Preparation of compound I-2

[0082]

[0083] Example 1 step (1), 1,4-dibromobutane (716.52 μL, 6 mmol) was used instead of 1,3-dibromopropane, and other conditions were unchanged to obtain compound A2 (white solid, 366.1 mg, yield about 65.8%).

[0084]

[0085] The preparation method of compound I-1 in step (3) of Example 1 was referred to, compound A2 was replaced by compound A1 in equal molar amount, and the target compound I-2 (orange red powder, 61.8 mg, yield 47.7%) was obtained without change of other conditions.

[0086] m.p.: 227.7 °C.

[0087] ESI-MS: 648.3883 [M+H] + .

[0088] 1 H-NMR (300 MHz, DMSO-d6, TMS), δ ppm: 8.73 (s, 1H), 7.08 (d, J = 6.7 Hz, 1H), 6.75 (d, J = 12.0 Hz, 1H), 6.40 (s, 1H), 6.36 (d, J = 7.0 Hz, 1H), 6.00 (d, J = 12.0 Hz, 1H), 4.06 (t, J = 6.0 Hz, 2H), 3.97-3.80 (m, 2H), 2.86 (s, 3H), 2.84 (s, 3H), 2.10 (s, 3H), 1.39 (s, 3H), 1.23 (s, 3H), 1.13 (s, 3H), 1.08 (s, 3H), 0.48 (s, 3H).

[0089] Example 3

[0090] Preparation of compound I-3

[0091]

[0092] The preparation method of compound I-1 in step (3) of Example 1 was referred to, compound A2 was replaced by compound A1 in equal molar amount, and the target compound I-2 (orange red powder, 61.8 mg, yield 47.7%) was obtained without change of other conditions.

[0093]

[0094] The preparation method of compound I-1 in step (3) of Example 1 was referred to, compound A2 was replaced by compound A1 in equal molar amount, and the target compound I-2 (orange red powder, 61.8 mg, yield 47.7%) was obtained without change of other conditions.

[0095] m.p.: 230.0 °C.

[0096] ESI-MS: 662.4032 [M+H] + .

[0097] 1H-NMR (300 MHz, DMSO-d6, TMS), δ ppm: 8.72 (s, 1H), 7.08 (d, J = 6.8 Hz, 1H), 6.75 (d, J = 12.0 Hz, 1H), 6.38 (s, 1H), 6.35 (d, J = 7.0 Hz, 1H), 6.00 (d, J = 12.0 Hz, 1H), 4.04 (t, J = 6.4 Hz, 2H), 3.94-3.77 (m, 2H), 2.85 (s, 3H), 2.83 (s, 3H), 2.10 (s, 3H), 1.39 (s, 3H), 1.23 (s, 3H), 1.13 (s, 3H), 1.08 (s, 3H), 0.47 (s, 3H).

[0098] Example 4

[0099] Preparation of compound I-4

[0100]

[0101] Step (1), Maleic anhydride (200 mg, 2.04 mmol) was dissolved in 5 mL of anhydrous dichloromethane, diethylamine (317 μL, 3.06 mmol) and triethylamine (425.3 μL, 3.06 mmol) were added successively, the reaction was stirred at room temperature, TLC was used to detect the reaction progress (dichloromethane:methanol = 20:1 V / V), after 1 h, the reaction was detected to be complete. After the reaction was completed, the organic phase was rotary evaporated to obtain (E)-4-(diethylamino)-4-oxobut-2-enoic acid, which was directly used in the next step.

[0102]

[0103] Step (2), all (E)-4-(diethylamino)-4-oxobut-2-enoic acid obtained in step (1), K2CO3 (552.84 mg, 4 mmol) were stirred and dissolved in 4 mL of DMF, 1,3-dibromopropane (609 μL, 6 mmol) was added, the reaction was carried out at room temperature, TLC was used to detect the reaction progress (petroleum ether: ethyl acetate = 1:1 V / V), after 4 h, the reaction was detected to be complete. After the reaction was completed, 90 mL of ethyl acetate was added to dilute the reaction solution, water washing (30 mL x 3) was carried out, the organic phase was taken, the organic phase was washed with saturated NaCl solution (100 mL x 3), anhydrous Na2SO4 was added for drying overnight, suction filtration was carried out, the filtrate was concentrated, normal phase silica gel column chromatography (eluent: petroleum ether: ethyl acetate = 3:1 V / V) was carried out, vacuum drying was carried out, and compound A4 (white solid, 338.2 mg, yield about 58.1%) was obtained.

[0104]

[0105] Step (3), gossypol (90.12 mg, 0.2 mmol), compound A4 (116.42 mg, 0.4 mmol) and NaHCO3(67.2 mg, 0.8 mmol) were dissolved in 4 mL of DMF, and the reaction was heated to reflux at 60 °C. TLC was used to monitor the reaction progress (dichloromethane:methanol = 50:1 V / V). After 8 h, the reaction was determined to be complete. After the reaction was completed, 90 mL of ethyl acetate was added to dilute the reaction solution, which was washed with water (30 mL x 3). The organic phase was washed with saturated NaCl solution (100 mL x 3), dried over anhydrous Na2SO4overnight, filtered, and concentrated. The target compound I-4 (orange red powder, 68.9 mg, yield 52.1%) was obtained by normal phase silica gel column chromatography (eluent dichloromethane:methanol = 150:1 V / V) and vacuum drying.

[0106] m.p.: 224.2 °C.

[0107] ESI-MS: 662.4034 [M+H] + .

[0108] 1 H-NMR (300 MHz, DMSO-d6, TMS), δ ppm: 8.73 (s, 1H), 7.09 (d, J = 6.2 Hz, 1H), 6.75 (d, J = 12.0 Hz, 1H), 6.39 (s, 1H), 6.35 (d, J = 6.4 Hz, 1H), 5.96 (d, J = 11.9 Hz, 1H), 4.11 (t, J = 6.6 Hz, 2H), 4.01-3.82 (m, 2H), 3.30-3.22 (m, 2H), 3.17 (q, J = 7.1 Hz, 2H), 2.10 (s, 3H), 1.39 (s, 3H), 1.23 (s, 3H), 1.13 (s, 3H), 1.08 (s, 3H), 1.02 (m, 6H), 0.46 (s, 3H).

[0109] Example 5

[0110] Preparation of compound I-5

[0111]

[0112] Compound A5 (white solid, 435.6 mg, yield about 71.4%) was obtained by replacing 1,3-dibromopropane in Example 4 step (2) with 1,4-dibromobutane, and other conditions were unchanged.

[0113]

[0114] The preparation method of compound I-4 in step (3) of example 4 was referred to, compound A5 was replaced by compound A4 in equimolar amount, and the target compound I-5 (orange red powder, 70 mg, yield 51.8%) was obtained without change of other conditions.

[0115] m.p.: 229.4 °C.

[0116] ESI-MS: 698.3995 [M+Na] + .

[0117] 1 H-NMR (300 MHz, DMSO-d6, TMS), δ ppm: 8.73 (s, 1H), 7.08 (d, J = 6.1 Hz, 1H), 6.81 (d, J = 12.0 Hz, 1H), 6.39 (s, 1H), 6.36 (d, J = 6.7 Hz, 1H), 5.98 (d, J = 11.9 Hz, 1H), 4.05 (t, J = 5.9 Hz, 2H), 3.96-3.79 (m, 2H), 3.29 (q, J = 7.1 Hz, 2H), 3.21 (q, J = 7.1 Hz, 2H), 2.10 (s, 3H), 1.39 (s, 3H), 1.22 (s, 3H), 1.13 (s, 3H), 1.08 (s, 3H), 1.04 (t, J = 6.8 Hz, 6H), 0.47 (s, 3H).

[0118] Example 6

[0119] Preparation of compound I-6

[0120]

[0121] The preparation method of compound I-4 in step (3) of example 4 was referred to, compound A5 was replaced by compound A4 in equimolar amount, and the target compound I-5 (orange red powder, 70 mg, yield 51.8%) was obtained without change of other conditions.

[0122]

[0123] The preparation method of compound I-4 in step (3) of example 4 was referred to, compound A5 was replaced by compound A4 in equimolar amount, and the target compound I-5 (orange red powder, 70 mg, yield 51.8%) was obtained without change of other conditions.

[0124] m.p.: 227.8 °C.

[0125] ESI-MS: 690.4352 [M+H] + .

[0126] 1H-NMR (300 MHz, DMSO-d6, TMS), δ ppm: 8.72 (s, 1H), 7.08 (d, J = 6.7 Hz, 1H), 6.81 (d, J = 12.0 Hz, 1H), 6.38 (s, 1H), 6.34 (d, J = 6.4 Hz, 1H), 5.99 (d, J = 12.0 Hz, 1H), 4.03 (t, J = 6.4 Hz, 2H), 3.86 (dp, J = 22.3, 5.6 Hz, 2H), 3.28 (q, J = 7.1 Hz, 2H), 3.20 (q, J = 7.0 Hz, 2H), 2.10 (s, 3H), 1.39 (s, 3H), 1.22 (s, 3H), 1.13 (s, 3H), 1.08 (s, 3H), 1.04 (t, J = 7.2 Hz, 6H), 0.47 (s, 3H). Example 7

[0127] Preparation of compound I-7

[0128]

[0129] Step (1), Maleic anhydride (200 mg, 2.04 mmol) was dissolved in 5 mL of anhydrous dichloromethane, diisopropylamine (429 μL, 3.06 mmol), triethylamine (425.3 μL, 3.06 mmol) were added successively, the reaction was stirred at room temperature, TLC was used to detect the reaction progress (dichloromethane:methanol = 20:1 V / V), after 1 h, the reaction was detected to be complete. After the reaction was completed, the organic phase was rotary evaporated to obtain (E)-4-(diisopropylamino)-4-oxobut-2-enoic acid, which was directly used in the next step.

[0130]

[0131] Step (2), all (E)-4-(diisopropylamino)-4-oxobut-2-enoic acid obtained in step (1), K2CO3 (552.84 mg, 4 mmol) were stirred and dissolved in 4 mL of DMF, 1,3-dibromopropane (609 μL, 6 mmol) was added, the reaction was carried out at room temperature, TLC was used to detect the reaction progress (petroleum ether: ethyl acetate = 2:1 V / V), after 4 h, the reaction was detected to be complete. After the reaction was completed, 90 mL of ethyl acetate was added to dilute the reaction solution, water washing (30 mL x 3) was carried out, the organic phase was taken, the organic phase was washed with saturated NaCl solution (100 ml x 3), dried with anhydrous Na2SO4 overnight, suction filtered, the filtrate was concentrated, purified by normal phase silica gel column chromatography (eluent was petroleum ether: ethyl acetate = 6:1 V / V), and dried in vacuum to obtain compound A7 (white solid, 383.7 mg, yield about 59.1%).

[0132]

[0133] Step (3), gttifern (90.12 mg, 0.2 mmol), compound A7 (127.6 mg, 0.4 mmol) and NaHC03(67.2 mg, 0.8 mmol) were dissolved in 4 mL of DMF, and the reaction was heated to reflux at 60 °C. The reaction progress was monitored by TLC (dichloromethane:methanol = 50:1 V / V). After 8 h, the reaction was determined to be complete. After the reaction was completed, 90 mL of ethyl acetate was added to dilute the reaction solution, which was washed with water (30 mL x 3). The organic phase was washed with saturated NaCl solution (100 mL x 3), dried over anhydrous Na2S04overnight, filtered, and concentrated. The target compound I-7 (orange red powder, 92.5 mg, yield 67.1%) was obtained by normal phase silica gel column chromatography (eluent dichloromethane:methanol = 150:1 V / V) and vacuum drying.

[0134] m.p.: 231.8 °C.

[0135] ESI-MS: 690.4343 [M+H] + .

[0136] 1 H-NMR (300 MHz, DMSO-d6, TMS), δ ppm: δ 8.73 (s, 1H), 7.09 (d, J = 6.7 Hz, 1H), 6.72 (d, J = 12.0 Hz, 1H), 6.39 (s, 1H), 6.36 (d, J = 7.0 Hz, 1H), 5.85 (d, J = 12.0 Hz, 1H), 4.17-4.07 (m, 2H), 4.02-3.82 (m, 2H), 3.75 (p, J = 6.5 Hz, 1H), 3.44 (p, J = 6.8 Hz, 1H), 2.10 (s, 3H), 1.39 (s, 3H), 1.32 (d, J = 6.7 Hz, 6H), 1.22 (s, 3H), 1.13 (s, 3H), 1.08 (s, 3H), 1.05 (d, J = 2.3 Hz, 3H), 1.03 (d, J = 2.3 Hz, 3H), 0.46 (s, 3H).

[0137] Example 8

[0138] Preparation of compound I-8

[0139]

[0140] With 1,4-dibromobutane (716.52 μL, 6 mmol) instead of 1,3-dibromopropane in Example 7 step (2), and other conditions unchanged, compound A8 (white solid, 434 mg, yield about 63.9%) was obtained.

[0141]

[0142] Referring to the preparation method of compound I-7 in step (3) of Example 7, compound A7 was replaced with an equimolar amount of compound A8, and other conditions remained unchanged, to obtain the target compound I-8 (orange-red powder, 100.3 mg, yield 71.3%).

[0143] mp:230.2℃.

[0144] ESI-MS: 726.4326 [M+Na] + .

[0145] 1 H-NMR (300MHz, DMSO-d6, TMS), δppm: 8.73 (s, 1H), 7.08 (d, J = 6.8Hz, 1H), 6.76 (d, J = 1 1.9Hz,1H),6.39(s,1H),6.35(d,J=6.4Hz,1H),5.87(d,J=11.9Hz,1H),4.10-4.04(m, 2H),3.93-3.78(m,3H),3.46(p,J=6.8Hz,1H),2.10(s,3H),1.39(s,3H),1.33(d,J=6 .7Hz,6H),1.22(s,3H),1.12(s,3H),1.09(s,3H),1.07(d,J=2.7Hz,6H),0.47(s,3H).

[0146] Example 9

[0147] Preparation of compound I-9

[0148]

[0149] Replacing 1,3-dibromopropane in step (2) of Example 7 with 1,5-dibromopentane (817 μL, 6 mmol), while keeping other conditions unchanged, yielded compound A9 (white solid, 401.6 mg, yield approximately 56.7%).

[0150]

[0151] Referring to the preparation method of compound I-7 in step (3) of Example 7, compound A7 was replaced with an equimolar amount of compound A9, and other conditions remained unchanged, to obtain the target compound I-9 (orange-red powder, 82.1 mg, yield 57.2%).

[0152] mp:227.3℃.

[0153] ESI-MS: 718.4660 [M+H] + .

[0154] 1 H-NMR (300 MHz, DMSO-d6, TMS), δ ppm: 8.73 (s, 1 H), 7.08 (d, J = 6.4 Hz, 1 H), 6.77 (d, J = 11.9 Hz, 1 H), 6.38 (s, 1 H), 6.35 (d, J = 6.7 Hz, 1 H), 5.88 (d, J = 12.0 Hz, 1 H), 4.05 (t, J = 6.4 Hz, 2 H), 3.92 - 3.75 (m, 3 H), 3.45 (p, J = 6.7 Hz, 1 H), 2.10 (s, 3 H), 1.39 (s, 3 H), 1.33 (d, J = 6.8 Hz, 6 H), 1.22 (s, 3 H), 1.12 (s, 3 H), 1.07 (d, J = 6.2 Hz, 9 H), 0.47 (s, 3 H).

[0155] Example 10

[0156] Preparation of compound 1-10

[0157]

[0158] Step (1), Maleic anhydride (200 mg, 2.04 mmol) was dissolved in 5 mL of anhydrous dichloromethane, morpholine (267 μL, 3.06 mmol), triethylamine (425.3 μL, 3.06 mmol) were added successively, the reaction was stirred at room temperature, TLC was used to monitor the reaction progress (dichloromethane:methanol = 20:1 V / V), after 1 h, the reaction was completed. After the reaction was completed, the organic phase was rotary evaporated to obtain the intermediate (E)-4-(morpholino)-4-oxobut-2-enoic acid, which was directly used in the next step.

[0159]

[0160] Step (2): Dissolve all of the (E)-4-(morpholino)-4-oxobut-2-enoic acid obtained in step (1) and K2CO3 (552.84 mg, 4 mmol) in 4 mL of DMF with stirring. Then add 1,3-dibromopropane (609 μL, 6 mmol) and react at room temperature. Monitor the reaction progress by TLC (petroleum ether: ethyl acetate = 1:2 V / V). After 4 h of reaction, the reaction was detected as complete. After the reaction was completed, add 90 mL of ethyl acetate to the reaction solution for dilution, wash with water (30 mL × 3), take the organic phase, wash the organic phase with saturated NaCl solution (100 mL × 3), dry with anhydrous Na2SO4 overnight, filter, concentrate the filtrate, and perform normal phase silica gel column chromatography (eluent: petroleum ether: ethyl acetate = 3:2 V / V). Dry under vacuum to obtain compound A10 (white solid, 313.8 mg, yield approximately 50.4%).

[0161]

[0162] Step (3): Tripterygium wilfordii (90.12 mg, 0.2 mmol), compound A10 (122 mg, 0.4 mmol), and NaHCO3 (67.2 mg, 0.8 mmol) were dissolved in 4 mL of DMF and heated to reflux at 60 °C. The reaction progress was monitored by TLC (dichloromethane:methanol = 42:1 V / V). After 8 h of reaction, the reaction was detected as complete. After the reaction was completed, 90 mL of ethyl acetate was added to the reaction solution for dilution, and the solution was washed with water (30 mL × 3). The organic phase was taken and washed with saturated NaCl solution (100 mL × 3). The solution was dried overnight with anhydrous Na2SO4, filtered, and the filtrate was concentrated. The solution was then subjected to normal phase silica gel column chromatography (eluent:dichloromethane:methanol = 126:1 V / V) and dried under vacuum to obtain the target compound I-10 (orange-red powder, 103.2 mg, yield 76.4%).

[0163] mp:227.9℃.

[0164] ESI-MS: 676.3831 [M+H] + .

[0165] 1H-NMR (300 MHz, DMSO-d6, TMS), δ ppm: 8.72 (s, 1 H), 7.09 (d, J = 6.7 Hz, 1 H), 6.70 (d, J = 12.0 Hz, 1 H), 6.39 (s, 1 H), 6.36 (d, J = 7.0 Hz, 1 H), 6.00 (d, J = 12.0 Hz, 1 H), 4.13 (dq, J = 15.0, 5.6 Hz, 2 H), 3.92 (ddt, J = 32.3, 11.4, 6.0 Hz, 2 H), 3.57 (t, J = 4.7 Hz, 2 H), 3.51 (t, J = 4.7 Hz, 2 H), 3.45 (t, J = 4.8 Hz, 2 H), 3.28 (t, J = 4.9 Hz, 2 H), 2.10 (s, 3 H), 1.39 (s, 3 H), 1.23 (s, 3 H), 1.14 (s, 3 H), 1.08 (s, 3 H), 0.47 (s, 3 H).

[0166] Example 11

[0167] Preparation of compound I-11

[0168]

[0169] With 1,4-dibromobutane (716.52 μL, 6 mmol) instead of 1,3-dibromopropane in Example 10 step (2), and other conditions unchanged, compound A11 (white solid, 367.8 mg, yield about 56.5%) was obtained.

[0170]

[0171] Referring to the preparation method of compound I-10 in Example 10 step (3), with equimolar amount of compound A11 instead of compound A10, and other conditions unchanged, the target compound I-11 (orange red powder, 74.4 mg, yield 54%) was obtained.

[0172] m.p.: 228.0 °C.

[0173] ESI-MS: 690.3998 [M+H] + .

[0174] 1H-NMR (300 MHz, DMSO-d6, TMS), δ ppm: 8.72 (s, 1 H), 7.08 (d, J = 6.8 Hz, 1 H), 6.76 (d, J = 11.8 Hz, 1 H), 6.40 (s, 1 H), 6.35 (d, J = 7.0 Hz, 1 H), 6.04 (d, J = 11.7 Hz, 1 H), 4.10 (t, J = 5.4 Hz, 2 H), 3.89 (dp, J = 17.5, 5.7 Hz, 2 H), 3.56 (t, J = 11.0, 4.6 Hz, 4 H), 3.46 (t, J = 4.6 Hz, 2 H), 3.29 (t, J = 5.1 Hz, 2 H), 2.10 (s, 3 H), 1.39 (s, 3 H), 1.24 (s, 3 H), 1.13 (s, 3 H), 1.08 (s, 3 H), 0.48 (s, 3 H). Example 12

[0175] Preparation of compound I-12

[0176]

[0177] With 1,5-dibromopentane (817 μL, 6 mmol) instead of 1,3-dibromopropane in Example 10 step (2), and other conditions unchanged, compound A12 (white solid, 384.5 mg, yield about 56.6%) was obtained.

[0178]

[0179] With compound A12 instead of compound A10 in equimolar amount, and other conditions unchanged, the target compound I-12 (orange red powder, 100.5 mg, yield 71.5%) was obtained according to the preparation method of compound I-10 in Example 10 step (3).

[0180] m.p.: 227.2 °C.

[0181] ESI-MS: 726.3956 [M+Na] + .

[0182] 1H-NMR (300 MHz, DMSO-d6, TMS), δ ppm: 8.72 (s, 1H), 7.08 (d, J = 7.0 Hz, 1H), 6.76 (d, J = 11.8 Hz, 1H), 6.38 (s, 1H), 6.35 (d, J = 6.7 Hz, 1H), 6.05 (d, J = 11.6 Hz, 1H), 4.06 (t, J = 6.3 Hz, 2H), 3.95-3.77 (m, 2H), 3.57 (t, J = 4.7 Hz, 2H), 3.53 (t, J = 4.2 Hz, 2H), 3.47 (t, J = 4.6 Hz, 2H), 3.29 (t, J = 5.5 Hz, 2H), 2.10 (s, 3H), 1.39 (s, 3H), 1.22 (s, 3H), 1.13 (s, 3H), 1.08 (s, 3H), 0.47 (s, 3H).

[0183] Example 13

[0184] Preparation of compound I-13

[0185]

[0186] Step (1), maleic anhydride (200 mg, 2.04 mmol) was dissolved in 5 mL of anhydrous dichloromethane, piperidine (280.2 μL, 3.06 mmol), triethylamine (425.3 μL, 3.06 mmol) were added successively, the reaction was stirred at room temperature, TLC was used to detect the reaction progress (dichloromethane:methanol = 20:1 V / V), after 1 h, the reaction was detected to be complete. After the reaction was completed, the organic phase was rotary evaporated to obtain the intermediate (E)-4-oxo-4-(piperidin-l-yl)but-2-enoic acid, which was subjected to the next step.

[0187]

[0188] Step (2), all the (E)-4-oxo-4-(piperidin-l-yl)but-2-enoic acid obtained in step (1), K2CO3 (552.84 mg, 4 mmol) were stirred and dissolved in 4 mL of DMF, 1,3-dibromopropane (609 μL, 6 mmol) was added, the reaction was carried out at room temperature, TLC was used to detect the reaction progress (petroleum ether: ethyl acetate = 1:1 V / V), after 4 h, the reaction was detected to be complete. After the reaction was completed, 90 mL of ethyl acetate was added to dilute the reaction solution, water washing (30 mL x 3) was carried out, the organic phase was taken, the organic phase was washed with saturated NaCl solution (100 mL x 3), dried over anhydrous Na2SO4 overnight, suction filtered, the filtrate was concentrated, and normal phase silica gel column chromatography (petroleum ether: ethyl acetate = 3:1 V / V) was carried out, and vacuum drying was carried out to obtain compound A13 (white solid, 405.7 mg, yield about 65.6%).

[0189]

[0190] Step (3), ginnalin (90.12 mg, 0.2 mmol), compound A13 (121.2 mg, 0.4 mmol) and NaHC03(67.2 mg, 0.8 mmol) were dissolved in 4 mL of DMF, and the reaction was heated to reflux at 60 °C. TLC was used to monitor the reaction progress (dichloromethane:methanol = 44:1 V / V). After 8 h, the reaction was determined to be complete. After the reaction was completed, 90 mL of ethyl acetate was added to dilute the reaction solution, which was washed with water (30 mL x 3). The organic phase was washed with saturated NaCl solution (100 mL x 3), dried over anhydrous Na2S04overnight, filtered, and concentrated. The target compound I-13 (orange red powder, 86.6 mg, yield 64.3%) was obtained by normal phase silica gel column chromatography (eluent dichloromethane:methanol = 132:1 V / V) and vacuum drying.

[0191] m.p.: 226.1 °C.

[0192] ESI-MS: 674.4038 [M+H] + .

[0193] 1 H-NMR (400 MHz, DMSO-d6, TMS), δ ppm: 8.75 (s, 1H), 7.09 (d, J = 6.8 Hz, 1H), 6.70 (d, J = 12.0 Hz, 1H), 6.38 (s, 1H), 6.35 (d, J = 6.4 Hz, 1H), 5.94 (d, J = 11.9 Hz, 1H), 4.17-4.06 (m, 2H), 4.01-3.81 (m, 2H), 3.41 (t, J = 5.7 Hz, 2H), 3.21 (t, J = 6.4 Hz, 2H), 2.10 (s, 3H), 1.39 (s, 3H), 1.22 (s, 3H), 1.13 (s, 3H), 1.07 (s, 3H), 0.46 (s, 3H).

[0194] Example 14

[0195] Preparation of compound I-14

[0196]

[0197] With 1,4-dibromobutane (716.52 μL, 6 mmol) instead of 1,3-dibromopropane in Example 13 step (2), and other conditions unchanged, compound A14 (white solid, 453.1 mg, yield about 70.1%) was obtained.

[0198]

[0199] Referring to the preparation method of compound I-13 in step (3) of Example 13, compound A13 was replaced with an equimolar amount of compound A14, and other conditions remained unchanged, to obtain the target compound I-14 (orange-red powder, 80.8 mg, yield 58.8%).

[0200] mp:222.6℃.

[0201] ESI-MS: 688.4196 [M+H] + .

[0202] 1 H-NMR (300MHz, DMSO-d6, TMS), δppm: 8.74 (s, 1H), 7.08 (d, J = 6.9 Hz, 1H), 6.76 (d, J =11.8Hz,1H),6.40(s,1H),6.36(d,J=6.5Hz,1H),5.97(d,J=11.8Hz,1H),4.06(t,J =5.9Hz,2H),3.88(dt,J=23.8,6.6Hz,2H),3.42(t,J=5.4Hz,2H),3.26(t,J=5.5Hz, 2H),2.10(s,3H),1.39(s,3H),1.22(s,3H),1.13(s,3H),1.08(s,3H),0.48(s,3H).

[0203] Example 15

[0204] Preparation of compound I-15

[0205]

[0206] Replacing 1,3-dibromopropane in step (2) of Example 13 with 1,5-dibromopentane (817 μL, 6 mmol), while keeping other conditions unchanged, yielded compound A15 (white solid, 330.8 mg, yield approximately 49%).

[0207]

[0208] Referring to the preparation method of compound I-13 in step (3) of Example 13, compound A13 was replaced with an equimolar amount of compound A15, and other conditions remained unchanged, to obtain the target compound I-15 (orange-red powder, 69.3 mg, yield 49.4%).

[0209] mp:224.9℃.

[0210] ESI-MS: 724.4163 [M+Na]+ .

[0211] 1 H-NMR (300 MHz, DMSO-d6, TMS), δ ppm: 8.74 (s, 1 H), 7.08 (d, J = 6.7 Hz, 1 H), 6.76 (d, J = 11.8 Hz, 1 H), 6.37 (s, 1 H), 6.34 (d, J = 7.0 Hz, 1 H), 5.98 (d, J = 11.8 Hz, 1 H), 4.04 (t, J = 6.2 Hz, 2 H), 3.95-3.76 (m, 2 H), 3.42 (t, J = 5.4 Hz, 2 H), 3.25 (t, J = 6.5 Hz, 2 H), 2.09 (s, 3 H), 1.39 (s, 3 H), 1.23 (s, 3 H), 1.13 (s, 3 H), 1.07 (s, 3 H), 0.47 (s, 3 H).

[0212] Example 16

[0213] Preparation of compound II-1

[0214]

[0215] Step (1), dissolve N-(tert-butoxycarbonyl)-1,2-ethanediamine (296.4 μL, 1.87 mmol) in 5 mL anhydrous dichloromethane, add triethylamine (780.8 μL, 5.62 mmol), stir at 0 °C for 5 min; then add acryloyl chloride (304.3 μL, 3.75 mmol), stir at room temperature, TLC monitor the reaction progress (petroleum ether: ethyl acetate = 1:2 V / V), after 1 h of reaction, it is detected that the reaction is complete. After the reaction is completed, the organic phase is rotary evaporated, 90 mL of ethyl acetate is added, washed with water (30 mL x 3), take the organic phase, the organic phase is washed with saturated NaCl solution (100 ml x 3), dried overnight with anhydrous Na2SO4, suction filtered, the filtrate is concentrated, and column chromatography is performed on normal phase silica gel (eluent petroleum ether: ethyl acetate = 3:2 V / V), vacuum dried to obtain A16-1 (white solid, 227.1 mg, yield 56.7%);

[0216] Step (2), dissolve compound A16-1 179.7 mg in 4 mL anhydrous CH2Cl2, drop 0.4 mL trifluoroacetic acid, react at room temperature for 1.5 h, TLC monitor at this time the reaction is completed. After the reaction is completed, the organic phase is rotary evaporated to obtain compound A16 (white sticky material, 95.8 mg, yield about 79.2%).

[0217]

[0218] Step (3), celastrol (90.12 mg, 0.2 mmol), HATU (114.1 mg, 0.3 mmol) were dissolved in 4 mL DMF, DIPEA (139.3 μL, 0.8 mmol) was added dropwise, stirred at room temperature until TLC detection showed that celastrol was activated, compound A16 (91.2 mg, 0.8 mmol) was added, and the reaction was continued at room temperature, and the reaction progress was monitored by TLC (dichloromethane:methanol = 25:1 V / V), and the reaction was complete after 4 h. After the reaction was completed, 90 mL of ethyl acetate was added to dilute the reaction solution, washed with water (30 mL x 3), and the organic phase was washed with saturated NaCl solution (100 mL x 3), dried over anhydrous Na2SO4 overnight, filtered, and the filtrate was concentrated, and the target compound II-1 (orange red powder, 32.2 mg, yield 29.5%) was obtained by normal phase silica gel column chromatography (eluent dichloromethane:methanol = 75:1 V / V) and vacuum drying.

[0219] m.p.: 200.6 °C.

[0220] ESI-MS: 547.3523 [M+H] + .

[0221] 1 H-NMR (300 MHz, DMSO-d6, TMS), δ ppm: 8.73 (s, 1H), 8.11 (t, J = 5.3 Hz, 1H), 7.65 (t, J = 4.9 Hz, 1H), 7.07 (d, J = 6.7 Hz, 1H), 6.39 (s, 1H), 6.34 (d, J = 7.0 Hz, 1H), 6.14 (dd, J = 17.1, 9.4 Hz, 1H), 6.04 (dd, J = 17.0, 2.9 Hz, 1H), 5.56 (dd, J = 9.4, 2.9 Hz, 1H), 3.15-2.92 (m, 4H), 2.09 (s, 3H), 1.37 (s, 3H), 1.20 (s, 3H), 1.07 (s, 3H), 1.04 (s, 3H), 0.51 (s, 3H).

[0222] Example 17

[0223] Preparation of compound II-2

[0224]

[0225] With N-(tert-butoxycarbonyl)-1,3-propanediamine (300.6 μL, 1.72 mmol) instead of N-(tert-butoxycarbonyl)-1,2-ethanediamine in Example 16 step (1), other conditions remain the same, to obtain compound A17 (white viscous substance, 108.8 mg, yield about 49.4%).

[0226]

[0227] Referring to the preparation method of compound II-1 in Example 16 step (3), compound A17 was replaced with an equimolar amount of compound A16, and other conditions were unchanged to obtain the target compound II-2 (orange red powder, 33.3 mg, yield 29.7%).

[0228] m.p.: 212.5 °C.

[0229] ESI-MS: 561.3672 [M+H] + .

[0230] 1 H-NMR (300 MHz, DMSO-d6, TMS), δ ppm: 8.74 (s, 1H), 8.04 (t, J = 5.5 Hz, 1H), 7.62 (t, J = 5.6 Hz, 1H), 7.08 (d, J = 6.8 Hz, 1H), 6.39 (s, 1H), 6.35 (d, J = 7.0 Hz, 1H), 6.16 (dd, J = 17.1, 9.8 Hz, 1H), 6.03 (dd, J = 17.1, 2.6 Hz, 1H), 5.53 (dd, J = 9.8, 2.6 Hz, 1H), 3.13-2.79 (m, 4H), 2.09 (s, 3H), 1.38 (s, 3H), 1.21 (s, 3H), 1.08 (s, 3H), 1.04 (s, 3H), 0.53 (s, 3H).

[0231] Example 18

[0232] Preparation of compound II-3

[0233]

[0234] Referring to the preparation method of compound II-1 in Example 16 step (3), compound A17 was replaced with an equimolar amount of compound A16, and other conditions were unchanged to obtain the target compound II-2 (orange red powder, 33.3 mg, yield 29.7%).

[0235]

[0236] Referring to the preparation method of compound II-1 in Example 16 step (3), compound A17 was replaced with an equimolar amount of compound A16, and other conditions were unchanged to obtain the target compound II-2 (orange red powder, 33.3 mg, yield 29.7%).

[0237] m.p.: 217.0 °C.

[0238] ESI-MS: 575.3828 [M+H] + .

[0239] 1 H-NMR (300 MHz, DMSO-d6, TMS), δ ppm: 8.73 (s, 1H), 8.02 (t, J = 5.6 Hz, 1H), 7.58 (t, J = 5.5 Hz, 1H), 7.08 (d, J = 6.7 Hz, 1H), 6.40 (s, 1H), 6.34 (d, J = 7.0 Hz, 1H), 6.16 (dd, J = 17.1, 9.9 Hz, 1H), 6.02 (dd, J = 17.1, 2.6 Hz, 1H), 5.52 (dd, J = 9.9, 2.6 Hz, 1H), 3.08 - 2.82 (m, 4H), 2.09 (s, 3H), 1.38 (s, 3H), 1.21 (s, 3H), 1.07 (s, 3H), 1.04 (s, 3H), 0.53 (s, 3H).

[0240] Example 19

[0241] Preparation of compound II-4

[0242]

[0243] With N-(tert-butoxycarbonyl)-1,6-hexanediamine (310.9 μL, 1.39 mmol) instead of N-(tert-butoxycarbonyl)-1,2-ethanediamine in Example 16, step (1), and other conditions unchanged, compound A19 (white sticky substance, 178.3 mg, yield about 75.5%) was obtained.

[0244]

[0245] With equal molar amount of compound A19 instead of compound A16, and other conditions unchanged, the target compound II-4 (orange red powder, 48.8 mg, yield 40.5%) was obtained according to the preparation method of compound II-1 in Example 16, step (3).

[0246] m.p.: 212.1 °C.

[0247] ESI-MS: 603.4142 [M+H] + .

[0248] 1H-NMR (300 MHz, DMSO-d6, TMS), δ ppm: 8.74 (s, 1H), 8.03 (t, J = 5.6 Hz, 1H), 7.55 (t, J = 6.0 Hz, 1H), 7.08 (d, J = 6.4 Hz, 1H), 6.40 (s, 1H), 6.35 (d, J = 6.6 Hz, 1H), 6.20 (dd, J = 17.1, 9.9 Hz, 1H), 6.05 (dd, J = 17.1, 2.5 Hz, 1H), 5.55 (dd, J = 9.9, 2.5 Hz, 1H), 3.07 (q, J = 6.6 Hz, 2H), 2.90 (dq, J = 13.8, 6.9 Hz, 2H), 2.09 (s, 3H), 1.38 (s, 3H), 1.21 (s, 3H), 1.07 (s, 3H), 1.04 (s, 3H), 0.53 (s, 3H).

[0249] Example 20

[0250] Preparation of compound II-5

[0251]

[0252] With tert-butyl [2-(2-aminoethoxy)ethyl]carbamate (293.3 μL, 1.47 mmol) instead of N-(tert-butoxycarbonyl)-1,2-ethanediamine in Example 16, step (1), and other conditions unchanged, compound A20 (white sticky substance, 156.2 mg, yield about 67.3%) was obtained.

[0253]

[0254] With equal molar amount of compound A20 instead of compound A16, and other conditions unchanged, the target compound II-5 (orange red powder, 53 mg, yield 44.9%) was obtained according to the preparation method of compound II-1 in Example 16, step (3).

[0255] m.p.: 204.9 °C.

[0256] ESI-MS: 591.3771 [M+H] + .

[0257] 1H-NMR (300 MHz, DMSO-d6, TMS), δ ppm: 8.70 (s, 1 H), 8.09 (t, J = 5.5 Hz, 1 H), 7.55 (t, J = 5.4 Hz, 1 H), 7.07 (d, J = 6.7 Hz, 1 H), 6.39 (s, 1 H), 6.34 (d, J = 6.9 Hz, 1 H), 6.24 (dd, J = 17.1, 10.0 Hz, 1 H), 6.07 (dd, J = 17.1, 2.4 Hz, 1 H), 5.57 (dd, J = 10.0, 2.4 Hz, 1 H), 3.38 (t, J = 5.3 Hz, 4 H), 3.28-3.22 (m, 2 H), 3.15-3.04 (m, 2 H), 2.09 (s, 3 H), 1.38 (s, 3 H), 1.21 (s, 3 H), 1.07 (s, 3 H), 1.05 (s, 3 H), 0.53 (s, 3 H).

[0258] Example 21

[0259] Preparation of compound II-6

[0260]

[0261] With tert-butyl N-[2-[2-(2-aminoethoxy)ethoxy]ethyl]carbamate (293.3 μL, 1.47 mmol) instead of N-(tert-butoxycarbonyl)-1,2-ethanediamine in Example 16, step (1 ), and other conditions unchanged, compound A20 (white sticky substance, 186.6 mg, yield about 76.5%) was obtained.

[0262]

[0263] With equal molar amount of compound A21 instead of compound A16 in the preparation method of compound II-1 in Example 16, step (3), and other conditions unchanged, the target compound II-6 (orange red powder, 71.2 mg, yield 56.2%) was obtained.

[0264] m.p.: 208.2 °C.

[0265] ESI-MS: 635.4045 [M+H] + .

[0266] 1H-NMR (300 MHz, DMSO-d6, TMS), δ ppm: 8.71 (s, 1H), 8.14 (t, 1H), 7.59 (t, J = 5.7 Hz, 1H), 7.08 (d, J = 6.6 Hz, 1H), 6.40 (s, 1H), 6.36 (d, J = 6.9 Hz, 1H), 6.24 (dd, J = 17.1, 10.0 Hz, 1H), 6.07 (dd, J = 17.0, 2.3 Hz, 1H), 5.56 (dd, J = 10.0, 2.3 Hz, 1H), 3.42 (t, J = 7.2 Hz, 8H), 3.26 (d, J = 5.5 Hz, 2H), 3.02 (d, J = 3.8 Hz, 2H), 2.09 (s, 3H), 1.38 (s, 3H), 1.21 (s, 3H), 1.07 (s, 3H), 1.04 (s, 3H), 0.54 (s, 3H).

[0267] Example 22

[0268] The anti-tumor activity of the gossypol acrylamide derivatives of the application was tested by tetrazolium blue colorimetric method (MTT method), and gossypol (CEL) and CDDO-Me (Bardoxolone methyl) were selected as positive control drugs. An appropriate amount of test compound, CEL and CDDO-Me was weighed, dissolved in DMSO to prepare a stock solution with a concentration of 10 mmol / L, and then diluted with culture medium to prepare a series of solutions with concentrations of 0.25, 0.5, 1, 2 and 4 μmol / L.

[0269] Instruments: super-clean workbench (SW-CJ-1FD, AIRTECH, Suqing Antai), constant-temperature CO2 incubator (3111, Thermo, USA), high-pressure sterilization pot (YXO.SG41.280, Shanghai Huaxian), centrifuge (SIGMA), multi-biology microscope (IX71, Olympus, Japan), multifunctional enzyme marker (POLARstar, Omega, USA).

[0270] Reagents: DMEM, 1640 culture medium (KeyGEN), fetal bovine serum (GIBCO), trypsin (SIGMA), DMSO (SIGMA).

[0271] Cell strains: human breast cancer cell strains MDA-MB-231, MCF-7, human gastric cancer cell strain BGC-823 and human colon cancer cell strain HT-29, wherein the BGC-823 cells were cultured with RPMI-1640 culture medium, and the MDA-MB-231, MCF-7 and HT-29 cells were cultured with DMEM (high-sugar) culture medium (all purchased from Jiangsu Keygen Biotech Co., Ltd.).

[0272] Method: The frozen cell line was recovered and placed in a constant temperature 37℃, 5% CO2 incubator, and the liquid was changed once a day. When it was in the exponential growth phase and in good condition, it was plated. 1 mL of 0.25% trypsin digestion solution was added, and the cells were digested for 1-2 min. Under a microscope, when the adherent cells were round and contracted, the digestion solution was removed, 1-2 mL of culture medium containing 10% fetal bovine serum was added to prepare a cell suspension, and the cells were counted. According to the number of cells per hole (5x10 3 4 μmol / L), 3 replicates were set for each drug concentration, and cultured for 48 h. MTT reagent (5 mg / mL) was added to the 96-well plate, 20 μL / well, and incubated for 4 h. The culture medium in the plate was removed, 100 μL of DMSO was added to each well, and the absorbance value of each well was detected by a multifunctional enzyme label instrument at a wavelength of 570 nm. The inhibition rate was calculated according to the following formula.

[0273] Cell inhibition rate % = [(blank control OD value - drug group OD value) / blank control group OD value] x 100%.

[0274] The average of 3 primary screening results is the final inhibition rate, and the IC 50 value of the test drug is calculated (graphpad software). The results of 3 repeated experiments are the final IC 50 value of the measured compound.

[0275] Table 1. Inhibition rate of test compound (concentration 0.5 μmol / L) on human breast cancer cell line MDA-MB-231

[0276]

[0277] Table 2. Inhibition of test compound on MDA-MB-231, MCF-7, BGC-823 and HT-29 cell lines

[0278]

[0279] Based on Table 1 and Table 2, compared with the emladin, the emladin acrylamide derivative of the application has significantly enhanced inhibition effect on MDA-MB-231, MCF-7, BGC-823 and HT-29 cell strains. Among them, the activity of compound I-4 is the best, and the activity is increased by about 5.3 times compared with the emladin, which is expected to become a new anti-tumor candidate, and is worthy of further research.

Claims

1. A triptolide acrylamide derivative with the structure shown in Formula III: in, A1 and A2 are independently selected from O and NH, respectively; L is selected from -(CH2) n -、-((CH2)2O) m (CH2)2-, where n is an integer from 2 to 6, and m is an integer from 1 to 6; R1 and R2 are independently selected from H, R3 and R4 are independently selected from C1-C4 alkyl groups, or R3 and R4 are connected to each other and together with oxygen atoms to form a 5- to 7-membered heterocycle containing one oxygen atom, or R3 and R4 are connected to each other and together with nitrogen atoms to form a 5- to 7-membered heterocycle.

2. The triptolide acrylamide derivative according to claim 1, characterized in that: A1 and A2 are each independently selected from O; L is selected from -(CH2) n -, where n is an integer from 3 to 5; R1 is selected from H, R2 is selected from R3 and R4 are independently selected from C1-C4 alkyl groups, or R3 and R4 are connected to each other and together with oxygen atoms to form a 5-7 membered heterocycle containing one oxygen atom, or R3 and R4 are connected to each other and together with nitrogen atoms to form a 5-7 membered heterocycle. Alternatively, A1 and A2 can be independently selected from NH; L is selected from -(CH2) n -、-((CH2)2O) m (CH2)2-, where n is an integer from 2 to 6 and m is an integer from 1 to 3; R1 and R2 are each independently selected from H.

3. The triptolide acrylamide derivative according to claim 2, characterized in that: A1 and A2 are each independently selected from O; L is selected from -(CH2) n -, where n is an integer from 3 to 5; R1 is selected from H, R2 is selected from R3 and R4 are independently selected from CH3, CH2CH3, and CH(CH3)2, respectively, or R3 and R4 are connected to each other and together with oxygen atoms to form a 6-membered heterocycle containing 1 oxygen atom, or R3 and R4 are connected to each other and together with nitrogen atoms to form a 6-membered heterocycle. Alternatively, A1 and A2 can be independently selected from NH; L is selected from -(CH2) n -、-(CH2)2O(CH2)2-,-(CH2)2O(CH2)2O(CH2)2-,n is an integer from 3 to 6; R1 and R2 are each independently selected from H; However, it does not include: A1 = A2 = O, L is selected from -(CH2)5-, R1 is selected from H, R2 is selected from R3=R4=CH3, R3=R4=CH(CH3)2, or R3 and R4 are connected to each other and together with nitrogen atoms to form a 6-membered heterocycle.

4. The triptolide acrylamide derivative according to claim 3, characterized in that: When A1 and A2 are independently selected from O, L is selected from -(CH2). n -, n is 3, R1 is selected from H, R2 is selected from R3 and R4 are each independently selected from CH3; L is selected from -(CH2). n - where n is an integer from 3 to 5, R1 is selected from H, and R2 is selected from H. R3 and R4 are each independently selected from CH2CH3; L is selected from -(CH2). n - where n is an integer from 3 to 5, R1 is selected from H, and R2 is selected from H. R3 and R4 are interconnected and together with the oxygen atom form a 6-membered heterocycle containing one oxygen atom; L is selected from -(CH2). n - where n is an integer from 3 to 4, R1 is selected from H, and R2 is selected from H. R3 and R4 are interconnected and together with nitrogen atoms form a 6-membered heterocycle; When A1 and A2 are independently selected from NH, L is selected from -(CH2). n -、-(CH2)2O(CH2)2-、-(CH2)2O(CH2)2O(CH2)2-, n is an integer from 4 to 6, and R1 and R2 are independently selected from H.

5. Selected from triptolide acrylamide derivatives with the structure shown below:

6. A method for preparing the triptolide acrylamide derivative, characterized in that: When A1 and A2 are independently selected from O, and L is selected from -(CH2). n - R1 is selected from H, R2 is selected from The synthesis route is as follows: Wherein, n, R3, and R4 are as described in claim 1; Includes the following steps: Step (1): Using anhydrous dichloromethane as the reaction solvent and triethylamine as the base catalyst, maleic anhydride reacts with the amine compound shown in Formula IV at room temperature to obtain intermediate V; Step (2): Using N,N-dimethylformamide as the reaction solvent and K2CO3 as the base catalyst, intermediate V reacts with... The dibromoalkane shown reacts at room temperature to give intermediate VI; Step (3): Using N,N-dimethylformamide as the reaction solvent and NaHCO3 as the base catalyst, intermediate VI reacts with triptolide under reflux to obtain triptolide acrylamide derivative. When A1 and A2 are independently selected from NH, and L is selected from -(CH2). n -、-((CH2)2O) m When (CH2)2-, R1 and R2 are independently selected from H, the synthetic route is as follows: Wherein, n, m, and L are as described in claim 1; Includes the following steps: Step (1): Using anhydrous dichloromethane as the reaction solvent and triethylamine as the base catalyst, the formula... The N-(tert-butyloxycarbonyl)diamine compound shown reacts with chloroacetamide of formula VII at room temperature to give intermediate VIII; Step (2): Using anhydrous dichloromethane as the reaction solvent, in the presence of trifluoroacetic acid, intermediate VIII is acid-hydrolyzed at room temperature to remove the Boc protecting group and obtain intermediate IX; Step (3): Using N,N-dimethylformamide as the reaction solvent, in the presence of HATU and DIPEA, intermediate IX undergoes an amidation reaction with the carboxyl group of triptolide at room temperature to obtain triptolide acrylamide derivative.

7. The method for preparing the triptolide acrylamide derivative according to claim 6, characterized in that: When A1 and A2 are independently selected from O, and L is selected from -(CH2). n - R1 is selected from H, R2 is selected from In step (1), the molar ratio of maleic anhydride to amine compound is 1:1.3 to 1:1.5; the molar ratio of maleic anhydride to triethylamine is 1:1.3 to 1:1.

5. In step (2), the molar ratio of intermediate V to dibromoalkane is 1:3 to 1:4; the molar ratio of intermediate V to K2CO3 is 1:2 to 1:

3. In step (3), the molar ratio of triptolide to intermediate VI is 1:1.5 to 1:2; the molar ratio of triptolide to NaHCO3 is 1:3 to 1:

4. When A1 and A2 are independently selected from NH, and L is selected from -(CH2). n -、-((CH2)2O) m When (CH2)2-, R1 and R2 are independently selected from H, in step (1), the molar ratio of the N-(tert-butoxycarbonyl)diamine compound to chloroacetamide is 1:1.5 to 1:2; the molar ratio of the N-(tert-butoxycarbonyl)diamine compound to triethylamine is 1:2 to 1:

3. In step (2), the volume ratio of trifluoroacetic acid to anhydrous dichloromethane is 1:5 to 1:10; In step (3), the molar ratio of triptolide to intermediate IX is 1:4 to 1:5; the molar ratio of triptolide to HATU is 1:1.2 to 1:1.5; and the molar ratio of triptolide to DIPEA is 1:3 to 1:

4.

8. Use of the triptolide acrylamide derivative according to any one of claims 1-5 in the preparation of an antitumor drug, wherein the tumor is breast cancer, gastric cancer, or colon cancer.

9. A pharmaceutical composition, characterized in that: It uses the triptolide acrylamide derivative as described in any one of claims 1-5 as the main active ingredient, supplemented with a pharmaceutically acceptable carrier, to formulate any pharmaceutically acceptable dosage form.

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