Majusculamide D derivative, preparation method and application of Majusculamide D derivative in preparation of anti-cancer drugs

Abstract

The invention provides a Majusculamide D derivative shown as a formula (I) or a pharmaceutically acceptable salt thereof, and the Majusculamide D derivative or the pharmaceutically acceptable salt thereof has a relatively good inhibition effect on various cancer cells and can be used for treating cancers. # imgabs0 #

Description

Technical Field

[0001] The invention provides a Majusculamide D derivative, relates to a preparation method of the Majusculamide D derivative, and use of the Majusculamide D derivative in preparing a drug for treating cancer. The invention belongs to the technical field of drugs. Background Art

[0002] Majusculamide D is a secondary metabolite extracted and isolated from the marine cyanobacteria Lyngbya majuscula in the Marshall Islands. Majusculamide D is a natural product of ester peptides with a chiral fatty side chain, multiple natural amino acids and α, β unsaturated ketone structures.

[0003] In 2019, Gerwick et al. completed the first total synthesis of Majusculamide D and found that it has good inhibitory activity against Panc-1 cells (IC 50 =0.32nM), which has the potential for further research and development. However, due to the low content of Majusculamide D in nature, its medicinal chemistry and mechanism have not been reported. The present invention provides the synthesis of Majusculamide D fatty side chain derivatives, and the derivatives have the effect of treating cancer and have IC 50 The values ​​range from 0.7 to 328.1 nM. Summary of the invention

[0004] The present invention provides a Majusculamide D derivative compound or a pharmaceutically acceptable salt thereof as shown in formula (I),

[0005]

[0006] In formula (I), R1 is hydrogen, methyl (R configuration) or methyl (S configuration); R2 is methyl (R configuration) or methyl (S configuration).

[0007] According to an embodiment of the present invention, the compound represented by formula (I) has the structure shown below:

[0008]

[0009] A method for preparing a Majusculamide D derivative represented by formula (I), characterized in that the compound represented by formula (II) is used as a raw material to prepare the compound represented by formula (I),

[0010]

[0011] A Majusculamide D derivative as shown in formula (I), and its use in preparing a cancer treatment or an auxiliary drug for cancer treatment, wherein the cancers are pancreatic cancer, lung adenocarcinoma, glioma, papillary lung adenocarcinoma, cervical cancer, human brain astrocytoma, liver cancer and colon cancer.

[0012] A pharmaceutical composition for treating cancer, comprising an effective amount of a Majusculamide D derivative of formula (I) in a pharmaceutically acceptable carrier or in combination with other anticancer drugs.

[0013] Beneficial Effects

[0014] The present invention provides a Majusculamide D derivative as shown in formula (I), which has good inhibitory effect on various cancer cells and can be used for treating cancer. DETAILED DESCRIPTION

[0015] In order to understand the present invention, the present invention is further illustrated by the following examples, but it is not intended to limit the protection scope of the present invention.

[0016] Example 1: Synthesis of Majusculamide D derivatives

[0017] The specific synthetic routes are shown in Route 1 and Route 2 below:

[0018]

[0019] Route 1

[0020]

[0021] Route 2

[0022] (1) Preparation of Compound 7

[0023] Compound 6 (10.0 g, 56.4 mmol) was added to a dry 500 mL three-necked round-bottom flask at room temperature, and dry THF (150 mL) was added to dissolve. The reaction system was replaced with argon three times, cooled to -78 °C, and n-BuLi (2.5 M in hexane, 22.6 mL, 56.4 mmol) was slowly added dropwise to the reaction system. After the addition was complete, the mixture was stirred for 2 h. Compound 5 (8.7 mL, 62.7 mmol) was then slowly added dropwise to the reaction system, and the mixture was stirred for 3 h at -78 °C. After the reaction was complete as detected by TLC, a saturated NH4Cl aqueous solution (100 mL) was added to the reaction system to quench the reaction. Ethyl acetate (100 mL) was added to dilute the reaction solution, and then the solution was allowed to stand for separation. The aqueous phase was extracted three times with ethyl acetate (3 × 100 mL), and the organic phases were combined. The organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 50 / 1-30 / 1) to obtain compound 7 (14.2 g, 91%) as a colorless oil. 1 HNMR (400MHz, CDCl3) δ7.38–7.18(m,5H),4.68(m,1H),4.24–4.13(m,2H),3.30(dd,J=13.3,3.1Hz,1H),3. 03–2.84(m,2H),2.77(dd,J=13.3,9.6Hz,1H),1.76–1.64(m,2H),1.45–1.30(m,4H),0.92(t,J=6.9Hz,3H); 13 C NMR (100MHz, CDCl3) δ173.61,153.61,135.49,129.57,129.10,127.49,66.30,55.31,38.10,35.65,31.43,24.11,22.59,14.07; HRMS(ESI)m / z:calcd for C 16 H 22 NO3 + [M+H] + :276.1594,found:276.1589.

[0024] (2) Preparation of Compound 8

[0025] Compound 7 (13.2 g, 47.9 mmol) was added to a dry 500 mL three-necked round-bottom flask at room temperature, and dry THF (200 mL) was added to dissolve. The reaction system was replaced with argon three times, cooled to -78 °C, and NaHMDS (2M in THF, 31.2 mL, 62.3 mmol) was slowly added dropwise to the reaction system. After the addition was complete, it was stirred for 1 h. Subsequently, CH3I (9.0 mL, 143.8 mmol) was slowly added dropwise to the reaction system, and stirring was continued for 3 h at -78 °C. After TLC detection of the reaction was complete, saturated NH4Cl aqueous solution (100 mL) was added to the reaction system to quench the reaction. Ethyl acetate (100 mL) was added to dilute the reaction solution, and then the solution was separated by standing. The aqueous phase was extracted with ethyl acetate (3×150 mL) three times, and the organic phases were combined. The organic phases were washed with saturated brine (100 mL), then dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 50 / 1-20 / 1) to obtain compound 8 (10.0 g, 72%) as a light yellow oil. 1 H NMR (400MHz, CDCl3) δ7.37–7.18(m,5H),4.68(m,1H),4.24–4.12(m,2H),3.71(q,J=6.8Hz,1H),3.27(dd,J=13.3,3.2Hz,1H), 2.77(dd,J=13.3,9.6Hz,1H),1.74(m,1H),1.46–1.38(m,1H),1.35–1.26(m,4H),1.22(d,J=6.8Hz,3H),0.89(t,J=6.9Hz,3H); 13 C NMR (100MHz, CDCl3) δ177.50,153.19,135.49,129.58,129.05,127.45,66.13, 55.48,38.04,37.80,33.27,29.55,22.85,17.49,14.09; HRMS(ESI)m / z:calcd for C 17 H 24 NO3 + [M+H] + :290.1751,found:290.1746.

[0026] (3) Preparation of Compound 9

[0027] Compound 8 (10.0 g, 34.6 mmol) was added to a dry 500 mL round-bottom flask at room temperature, and dry THF (100 mL) was added to dissolve. The temperature was lowered to 0°C, and NaBH4 (1.96 g, 51.8 mmol) was slowly added to the reaction system in batches, and stirred at 0°C. NaBH4 (1.30 g, 34.6 mmol) was slowly added again in batches after 2 h and 4 h, respectively, and then the reaction was warmed to room temperature and stirred overnight. After TLC detection of the reaction was complete, saturated NH4Cl aqueous solution (100 mL) was slowly added to the reaction system at 0°C to quench the reaction, and then the liquid was separated by standing. The aqueous phase was extracted 3 times with ethyl acetate (3×50 mL), and the organic phases were combined, washed with saturated brine (50 mL), and then dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 50 / 1-20 / 1) to obtain compound 9 (2.7 g, 68%) as a colorless oil. 1 H NMR (400MHz, CDCl3) δ3.51(m,1H),3.42(m,1H),1.64–1.57(m,1H),1.43–1.34(m,1H) ,1.33–1.25(m,4H),1.17–1.06(m,1H),0.92(d,J=6.7Hz,3H),0.89(t,J=6.7Hz,3H); 13 CNMR(100MHz, CDCl3)δ68.59,35.91,32.99,29.35,23.12,16.73,14.22; HRMS(ESI)m / z:calcd for C7H 16 ON + [M+Na] + :139.1093, compound 9 may be completely broken into fragments during the test process, and no molecular ion peak was detected.

[0028] (4) Preparation of Compound 10

[0029] PPh3 (5.25 g, 20.0 mmol) was added to a 250 mL round-bottom flask at room temperature, and DCM (50 mL) was added to dissolve. The temperature was lowered to 0 °C, I2 (5.5 g, 21.7 mmol) and imidazole (1.36 g, 20.0 mmol) were added to the reaction system, and stirred for 20 min. Compound 9 (1.8 g, 15.5 mmol) dissolved in DCM (10 mL) was then slowly added dropwise to the reaction system, and the reaction was warmed to room temperature and stirred for 3 h. After TLC detection of the reaction was complete, saturated Na2S2O3 aqueous solution (50 mL) was slowly added to the reaction system to quench the reaction, and then the reaction was allowed to stand for separation. The aqueous phase was extracted 3 times with DCM (3×50 mL), and the organic phases were combined, washed with saturated brine (50 mL), and then dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by silica gel column chromatography (n-pentane) to obtain compound 10 (2.1 g, 60%) as a light yellow oil. 1 H NMR (400MHz, CDCl3) δ3.23 (dd, J=9.5, 4.6Hz, 1H), 3.16 (dd, J=9.5, 5.9Hz, 1H) ,1.44(m,1H),1.36–1.17(m,6H),0.97(d,J=6.5Hz,3H),0.90(t,J=6.9Hz,3H); 13 C NMR (100MHz, CDCl3) δ36.29, 34.86, 29.27, 22.87, 20.76, 18.26, 14.21; HRMS (ESI) m / z:calcd for C7H 15 INa + [M+Na] + :249.0111, compound 10 may be completely broken into fragments during the test process, and no molecular ion peak was detected.

[0030] (5) Preparation of Compound 12

[0031] At room temperature, diisopropylamine (800 μL, 5.65 mmol) was added to a dry 25 mL three-necked round-bottom flask, and dry THF (1 mL) was added to dissolve. The reaction system was replaced with argon three times, cooled to -78°C, and n-BuLi (2.4 M in hexane, 2.25 mL, 5.41 mmol) was slowly added dropwise to the reaction system. After the addition was complete, the mixture was heated to room temperature and stirred for 1 h. Then the temperature was lowered to -78°C, and compound 11 (386 mg, 2.46 mmol) dissolved in THF (2 mL) was slowly added dropwise to the reaction system. After the addition was complete, the mixture was stirred for 1 h. HMPA (980 μL, 5.65 mmol) was added and stirred for 1 h. Then compound 10 (500 mg, 2.21 mmol) dissolved in THF (1 mL) was slowly added dropwise. After the addition was complete, the mixture was heated to -40°C and stirred for 4 h. After TLC detected that the reaction was complete, saturated NH4Cl aqueous solution (25 mL) was added to the reaction system to quench the reaction. Ethyl acetate (10 mL) was added to dilute the reaction solution, and then the mixture was allowed to stand for separation. The aqueous phase was extracted 3 times with ethyl acetate (3×20 mL), and the organic phases were combined. The organic phases were washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by preparative thin layer chromatography (dichloromethane / methanol=30 / 1) to obtain compound 12 (346 mg, 61%) as a light yellow oil. 1 H NMR (400MHz, CDCl3) δ5.29(s,1H),4.25(m,1H),3.57(m,4H),2.66(m,1H),2.04(m,1H),1.91(m,2H),1.72(m,1H),1.60(m,1 H),1.43–1.35(m,1H),1.28(m,5H),1.14(d,J=6.8Hz,4H),1.10–1.01(m,1H),0.89(d,J=6.8Hz,3H),0.86(d,J=6.6Hz,3H); 13 C NMR (100MHz, CDCl3) δ178.65,67.97,61.12,47.95,41.26,36.75,35.81,30.73,29.19,28.39,24.61,23.12,20.13,18.40,14.25; HRMS (ESI) m / z:calcd for C 15 H 30 NO2 + [M+H] + :256.2271,found:256.2268.

[0032] (6) Preparation of Compound 2

[0033] Compound 12 (2.68 g, 10.5 mmol) was added to a 10 mL round-bottom flask, and aqueous hydrochloric acid solution (1N, 52.5 mL, 52.5 mmol) was added to dissolve, and heated under reflux for 8 h. After TLC detection, ethyl acetate (50 mL) was added to the reaction system to dilute the reaction solution, and then the solution was allowed to stand for separation. The aqueous phase was extracted 3 times with ethyl acetate (3×50 mL), and the organic phases were combined, washed with saturated brine (100 mL), and then dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 20 / 1-10 / 1) to obtain a light yellow oil compound 2 (1.1 g, 62%). 1 H NMR(400MHz, CDCl3)δ2.62–2.50(m,1H),1.73(ddd,J=14.1,9.0,5.5Hz,1H),1.52–1.42(m,1H) ,1.28(dd,J=10.8,4.5Hz,5H),1.18(d,J=6.9Hz,3H),1.15–1.07(m,2H),0.89(t,J=6.1Hz,6H); 13 C NMR(100MHz, CDCl3)δ184.23,41.38,37.51,36.83,30.81,29.12,23.11,19.69,17.94,14.25; HRMS(ESI)m / z:calcd forC 10 H 19 O2 - [MH] - :171.1391,found:171.1387.

[0034] (7) Preparation of compound C40-epi-12

[0035] The synthesis method is the same as that of compound 12, except that the chiral auxiliary group 11 is replaced by ent-11. The crude product is separated and purified by preparative thin layer chromatography (dichloromethane / methanol = 30 / 1) to obtain a light yellow oil compound C40-epi-12 (310 mg, 55%). 1H NMR (400MHz, CDCl3) δ5.31(s,1H),4.27–4.16(m,1H),3.67–3.43(m,4H),2.61(m,1H),2.07–1.98(m,1H),1.89(m,1H),1. 57(m,1H),1.39(d,J=6.5Hz,3H),1.29–1.16(m,5H),1.10(d,J=6.7Hz,4H),0.87(d,J=7.0Hz,3H),0.84(d,J=6.1Hz,3H); 13 C NMR (100MHz, CDCl3) δ178.66,67.72,61.16,47.85,40.67,37.18,35.77,30.41,29.18,28.33,24.53,22.97,19.58,17.29,14.14; HRMS(ESI)m / z:calcd forC 15 H 30 NO2 + [M+H] + :256.2277,found:256.2268.

[0036] (8) Preparation of compound C40-epi-2

[0037] The synthesis method is the same as that of compound 2. The crude product was separated and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 40 / 1-20 / 1) to obtain compound C40-epi-2 (300 mg, 64%) as a light yellow oil. 1 H NMR (400MHz, CDCl3) δ2.54(m,1H),1.54(dt,J=12.3,7.1Hz,1H),1.47(dt,J=12.5,6.1Hz,1H),1. 42–1.35(m,1H),1.29(m,5H),1.16(d,J=6.9Hz,4H),0.89(d,J=5.9Hz,3H),0.86(d,J=6.3Hz,3H); 13 C NMR(100MHz, CDCl3)δ183.95,40.94,37.29,36.83,30.56,29.23,23.09,19.48,16.96,14.25; HRMS(ESI)m / z:calcd forC 10 H 19 O2 - [MH] - :171.1385,found:171.1386.

[0038] (9) Preparation of compound ent-7

[0039] The synthesis method is the same as that of compound 7, except that the chiral auxiliary group 6 is replaced by ent-6. The crude product is separated and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 50 / 1-30 / 1) to obtain a colorless oil compound ent-7 (73.6 g, 95%). 1 H NMR (400MHz, CDCl3) δ7.48–7.08(m,5H),4.68(m,1H),4.25–4.13(m,2H),3.30(dd,J=13.4,3.3Hz,1H),2. 93(m,2H),2.77(dd,J=13.3,9.6Hz,1H),1.75–1.66(m,2H),1.37(m,J=3.8Hz,4H),0.93(t,J=6.7Hz,3H); 13 C NMR (100MHz, CDCl3) δ173.45,153.50,135.39,129.48,128.97,127.35,66.18,55.17,37.93,35.53,31.32,23.98,22.50,14.00; HRMS(ESI)m / z:calcd for C 16 H 22 NO3 + [M+H] + :276.1594,found:276.1591.

[0040] (10) Preparation of compound ent-8

[0041] The synthesis method is the same as the method for preparing compound 8. The crude product was separated and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 40 / 1-20 / 1) to obtain a light yellow oily compound ent-8 (63.0 g, 82%). 68.9 (c = 1.0, CHCl3); 1 H NMR(400MHz, CDCl3)δ7.37–6.83(m,5H),4.65(m,1H), 3.69(q,J=6.5Hz,1H),3.25(d,J=13.4Hz,1H),2.81–2.70(m,1H),1.73(m,1H) ,1.40(m,1H),1.32–1.25(m,4H),1.21(d,J=5.9Hz,3H),0.89(t,J=5.8Hz,3H); 13C NMR (100MHz, CDCl3) δ177.44,153.15,135.46,129.55,129.01,127.41,66.09, 55.44,37.99,37.76,33.23,29.52,22.82,17.46,14.07; HRMS(ESI)m / z:calcd for C 17 H 24 NO3 + [M+H] + :290.1751,found:290.1748.

[0042] (11) Preparation of compound ent-9

[0043] The synthesis method is the same as the method for preparing compound 9. The crude product was separated and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 50 / 1-20 / 1) to obtain a colorless oil compound ent-9 (14.7 g, 69%). 1 H NMR (400MHz, CDCl3) δ3.50(m,1H),3.40(m,1H),1.68–1.52(m,1H),1.41–1.36(m, 1H),1.35–1.20(m,4H),1.09(m,1H),0.91(d,J=3.4Hz,3H),0.89(t,J=3.7Hz,3H); 13 C NMR (100MHz, CDCl3) δ68.48, 35.86, 32.98, 29.32, 23.09, 16.69, 14.18; HRMS (ESI) m / z:calcd for C7H 16 ON + [M+Na] + :139.1093, compound ent-9 may be completely broken into fragments during the test process, and no molecular ion peak was detected.

[0044] (12) Preparation of compound ent-10

[0045] The synthesis method is the same as the method for preparing compound 10. The crude product was separated and purified by silica gel column chromatography (n-pentane) to obtain a light yellow oily compound ent-10 (19.8 g, 69%). 1H NMR (400MHz, CDCl3) δ3.27–3.17(m,1H),3.20–3.11(m,1H),1.45(m,1H),1.40–1.14(m,6H),0.97(d,J=6.5Hz,3H),0.90(t,J=6.9Hz,3H); 13 C NMR (100MHz, CDCl3) δ36.30, 34.88, 29.28, 22.88, 20.76, 18.22, 14.20; HRMS (ESI) m / z:calcd for C7H 15 INa + [M+Na] + :249.0111, compound ent-10 may be completely broken into fragments during the test process, and no molecular ion peak was detected.

[0046] (13) Preparation of compound C42-epi-12

[0047] The synthesis method is the same as that of compound 12. The crude product was separated and purified by preparative thin layer chromatography (dichloromethane / methanol=30 / 1) to obtain compound C42-epi-12 (416 mg, 74%) as a light yellow oil. 1 H NMR (400MHz, CDCl3) δ5.31(s,1H),4.22(m,1H),3.70–3.45(m,4H),2.62(m,1H),2.09–1.79(m,3H),1.57(m ,1H),1.40(m,3H),1.30–1.21(m,5H),1.11(d,J=6.7Hz,4H),0.88(d,J=6.0Hz,3H),0.85(d,J=6.1Hz,3H); 13 C NMR (100MHz, CDCl3) δ178.77,67.84,61.22,47.90,40.68,37.21,35.79,30.41,29.21,28.38,24.57,23.01,19.60,17.29,14.20; HRMS (ESI) m / z:calcd for C 15 H 30 NO2 + [M+H] + :256.2271,found:256.2267.

[0048] (14) Preparation of compound C42-epi-2

[0049] The synthesis method is the same as that of compound 2. The crude product was separated and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 40 / 1-20 / 1) to obtain compound C42-epi-2 (750 mg, 56%) as a light yellow oil. 1 H NMR (400MHz, CDCl3) δ2.54(m,1H),1.51(m,2H),1.42–1.34(m,1H),1.33–1.20(m,5H),1.15(d,J=7.0Hz,4H),0.87(t,J=6.4Hz,6H); 13 C NMR(100MHz, CDCl3)δ183.79,40.91,37.25,36.82,30.52,29.22,23.09,19.46,16.95,14.27; HRMS(ESI)m / z:calcd for C 10 H 19 O2 - [MH] - 171.1391, found:171.1386.

[0050] (15) Preparation of compound C40-epi-C42-epi-12

[0051] The synthesis method is the same as that of compound 12, except that the chiral auxiliary group 11 is replaced by ent-11. The crude product is separated and purified by preparative thin layer chromatography (dichloromethane / methanol=30 / 1) to obtain a light yellow oil compound C40-epi-C42-epi-12 (317 g, 56%). 1 H NMR (400MHz, CDCl3) δ5.27(d,J=8.2Hz,1H),4.26(m,1H),3.70–3.45(m,4H),2.73–2.60(m,1H),1.97(m,3H),1.72(m,1H),1.59(m,1H) ,1.39(q,J=9.6,8.2Hz,1H),1.34–1.23(m,5H),1.14(d,J=6.7Hz,4H),1.10–1.02(m,1H),0.89(d,J=6.9Hz,3H),0.86(d,J=6.5Hz,3H); 13C NMR (100MHz, CDCl3) δ178.63,67.91,61.09,47.94,41.26,36.74,35.81,30.73,29.18,28.37,24.60,23.11,20.12,18.39,14.24; HRMS(ESI)m / z:calcd for C 15 H 30 NO2 + [M+H] + :256.2271,found:256.2268.

[0052] (16) Preparation of Compound C40-epi-C42-epi-2

[0053] The synthesis method is the same as that of compound 2. The crude product was separated and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 40 / 1-20 / 1) to obtain compound C40-epi-C42-epi-2 (630 mg, 55%) as a light yellow oil. 1 H NMR(400MHz, CDCl3) δ2.57(m,1H),1.73(ddd,J=14.0,9.0,5.4Hz,1H),1.51–1.40(m,1 H),1.32–1.22(m,5H),1.18(d,J=6.9Hz,3H),1.14–1.02(m,2H),0.89(t,J=6.5Hz,6H); 13 C NMR(100MHz, CDCl3)δ183.66,41.40,37.45,36.82,30.81,29.12,23.10,19.71,17.96,14.26; HRMS(ESI)m / z:calcd for C 10 H 19 O2 - [MH] - :171.1385,found:171.1385.

[0054] (17) Preparation of compound 7b

[0055] The synthesis method is the same as that of compound 7, except that compound 5 is replaced by 5b. The crude product is separated and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 40 / 1-10 / 1) to obtain compound 7b (7.84 g, 92%) as a colorless oil. 1H NMR (400MHz, CDCl3) δ7.33(t,J=7.3Hz,2H),7.30–7.25(m,1H),7.21(d,J=7.3Hz,2H),4.67(m,1H),4.24–4.12(m,2 H),3.29(d,J=13.2Hz,1H),2.93(m,2H),2.77(dd,J=13.3,9.8Hz,1H),1.69(m,2H),1.32(m,8H),0.92–0.86(m,3H); 13 C NMR (100MHz, CDCl3) δ173.56,153.58,135.45,129.54,129.06,127.44,66.25,55. 26,38.03,35.66,31.81,29.20,29.18,24.37,22.74,14.22; HRMS(ESI)m / z:calcd forC 18 H 26 NO3 + [M+H] + :304.1907,found:304.1906.

[0056] (18) Preparation of compound 8b

[0057] The synthesis method is the same as the method for preparing compound 8. The crude product was separated and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 40 / 1-10 / 1) to obtain a light yellow oil compound 8b (5.9 g, 81%). 1 H NMR (400MHz, CDCl3) δ7.36–7.27(m,3H),7.21(d,J=7.3Hz,2H),4.68(m,1H),4.27–4.13(m,2H),3.71(m,1H),3.27(dd,J=13.4,3 .3Hz,1H),2.77(dd,J=13.3,9.6Hz,1H),1.73(m,1H),1.46–1.36(m,1H),1.28(m,8H),1.22(d,J=6.7Hz,3H),0.91–0.83(m,3H); 13 C NMR (100MHz, CDCl3) δ177.39,153.07,135.37,129.45,128.93,127.33,66.01,55.37, 37.94,37.72,33.45,31.70,29.31,27.22,22.59,17.36,14.06; HRMS(ESI)m / z:calcd for C19 H 28 NO3 + [M+H] + :318.2064,found:318.2061.

[0058] (19) Preparation of compound 2b

[0059] Compound 8b (1.00 g, 3.15 mmol) was added to a 100 mL round-bottom flask, and THF (20 mL) and water (5 mL) were added to dissolve. The temperature was lowered to 0 ° C, and 30% H2O2 (9.8 M in H2O, 2.57 mL, 25.2 mmol) and LiOH (4.0 M in H2O, 3.15 mL, 12.6 mmol) were slowly added to the reaction system, and then the reaction was warmed to room temperature and stirred for 1.5 h. After TLC detection of the reaction was complete, the reaction solution was concentrated, and water (50 mL) was added to dilute the reaction solution. The aqueous phase was adjusted to pH 2 with 1N HCl aqueous solution, and then extracted with ethyl acetate (3×50 mL) for 3 times. The organic phases were combined, washed with saturated brine (200 mL), and then dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was separated and purified by silica gel column chromatography (dichloromethane / methanol=200 / 1-50 / 1) to obtain compound 2b (370 mg, 74%) as a light yellow oil. 1 H NMR (400MHz, CDCl3) δ2.45(m,1H),1.68(m,1H),1.43(m,1H),1.28(m,8H),1.17(dd,J=6.9,2.0Hz,3H),0.88(t,J=6.1Hz,3H); 13 C NMR(100MHz, CDCl3)δ184.00,39.60,33.69,31.82,29.32,27.25,22.74,16.91,14.14; HRMS(ESI)m / z:calcd for C9H 17 O2 - [MH] - :157.1234,found:157.1228.

[0060] (20) Preparation of compound 7c

[0061] The synthesis method is the same as that of compound 7b. The crude product is separated and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 40 / 1-10 / 1) to obtain compound 7c (8.29 g, 97%) as a colorless oil. 1H NMR (400MHz, CDCl3) δ7.37–7.26(m,3H),7.24–7.18(m,2H),4.73–4.62(m,1H),4.24–4.13(m,2H),3.30(dd,J=13. 3,3.4Hz,1H),3.04–2.83(m,2H),2.76(dd,J=13.3,9.6Hz,1H),1.69(m,2H),1.44–1.24(m,8H),0.93–0.83(m,3H); 13 C NMR (100MHz, CDCl3) δ173.59,153.60,135.46,129.56,129.08,127.47,66.27,55. 29,38.05,35.68,31.82,29.22,29.20,24.39,22.76,14.24; HRMS(ESI)m / z:calcd for C 18 H 26 NO3 + [M+H] + :304.1907,found:304.1905.

[0062] (21) Preparation of compound 8c

[0063] The synthesis method is the same as the method for preparing compound 8. The crude product was separated and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 40 / 1-10 / 1) to obtain a light yellow oil compound 8b (6.5 g, 81%). 1 H NMR (400MHz, CDCl3) δ7.31(m,3H),7.22(d,J=7.3Hz,2H),4.68(m,1H),4.19(m,2H),3.70(m,1H),3.27(dd,J=13.2,3.2Hz ,1H),2.77(dd,J=13.3,9.7Hz,1H),1.74(m,1H),1.41(m,1H),1.28(m,8H),1.22(d,J=6.9Hz,3H),0.87(t,J=6.5Hz,3H); 13 C NMR (100MHz, CDCl3) δ177.37,153.08,135.37,129.48,128.94,127.34,66.01,55.37, 37.90,37.73,33.45,31.72,29.33,27.24,22.62,17.39,14.11; HRMS(ESI)m / z:calcd forC 19 H28 NO3 + [M+H] + :318.2064,found:318.2061.

[0064] (22) Preparation of compound 2c

[0065] The synthesis method is the same as that of compound 2b. The crude product is separated and purified by silica gel column chromatography (dichloromethane / methanol=200 / 1-50 / 1) to obtain compound 2c (400 mg, 80%) as a light yellow oil. 1 H NMR (400MHz, CDCl3) δ2.46 (m, 1H), 1.68 (m, 1H), 1.42 (m, 1H), 1.28 (m, 8H), 1.17 (d, J = 6.9Hz, 3H), 0.88 (t, J = 6.6Hz, 3H); 13 C NMR(100MHz, CDCl3)δ183.98,39.65,33.71,31.82,29.32,27.25,22.74,16.95,14.16; HRMS(ESI)m / z:calcd forC9H 17 O2 - [MH] - :157.1234,found:157.1228.

[0066] (23) Preparation of Compound 1

[0067] Compound 13 (50 mg, 0.066 mmol) was added to a 10 mL round-bottom flask at room temperature, and THF (500 μL) was added to dissolve. The temperature was lowered to 0°C, and a hydrochloric acid dioxane solution (4N, 500 μL, 1.98 mmol) was added to the reaction system. The reaction was warmed to room temperature and stirred for 1 h. After TLC detected that the reaction was complete, the reaction solution was concentrated, and then an oil pump was used to spin off the high-boiling dioxane to obtain a white solid, which was directly used in the next step of the reaction.

[0068] Compound 2 (34 mg, 0.20 mmol) and HATU (75 mg, 0.20 mmol) were added to the reaction system at room temperature, and DCM (200 μL) was added to dissolve. The temperature was lowered to 0°C, and DIPEA (65 μL, 0.39 mmol) was slowly added dropwise to the reaction system. The reaction was warmed to room temperature and stirred overnight. After TLC detection of the reaction was complete, the reaction solution was concentrated, and ethyl acetate (5 mL) was added to dilute the reaction solution. The organic phase was first washed 3 times with 1N HCl (3×5 mL), then washed with saturated brine (5 mL), and then dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was separated and purified by silica gel column chromatography (dichloromethane / ethyl acetate = 2 / 1-1 / 1) and preparative thin layer chromatography (ethyl acetate) to obtain an off-white solid compound 1 (26 mg, 49%). 1 H NMR (400MHz, CDCl3) δ7.28(d,J=2.0Hz,1H),7.09(d,J=8.5Hz,2H),7.03(d,J=8.9Hz,1H),6.77(d, J=8.6Hz,1H),6.08(dd,J=6.1,1.6Hz,1H),5.66(dd,J=10.1,2.0Hz,1H),5.57(dd,J=11.2,5.6Hz, 1H),5.25(qd,J=6.4,2.7Hz,1H),5.00(d,J=11.1Hz,1H),4.96(dd,J=9.0,2.7Hz,1H),4.85–4.76( m,1H),4.37(s,1H),3.87(d,J=11.6Hz,1H),3.81(dd,J=11.6,4.4Hz,1H),3.75(s,3H),3.52(d,J=1 1.0Hz,1H),3.15(dd,J=15.4,5.9Hz,1H),3.08(s,3H),3.02–2.94(m,1H),2.92(s,3H),2.68–2.59 (m,1H),2.46(ddd,J=14.5,10.1,4.7Hz,1H),2.25(m,1H),2.02(d,J=14.1Hz,1H),1.98(s,3H),1.6 4(t,J=9.3Hz,1H),1.46(d,J=6.7Hz,3H),1.29–1.21(m,4H),1.16(d,J=6.5Hz,5H),1.06(d,J=6.7 Hz,3H),0.99(d,J=6.5Hz,3H),0.88(t,J=7.1Hz,5H),0.81(d,J=6.7Hz,3H),0.49(d,J=5.8Hz,3H); 13C NMR (100MHz, CDCl3) δ178.01,174.63,170.77,169.93,169.80,169.65,168.98,158.34 ,154.25,129.74,128.79,125.41,113.91,71.88,68.50,59.25,58.70,58.19,57.01,56 .44,55.17,51.96,41.67,37.28,36.70,33.49,32.44,30.69,30.46,30.36,29.08,27.2 2,23.12,21.12,19.46,18.91,18.58,18.21,17.49,17.02,14.24; HRMS(ESI)m / z:calcd forC 43 H 65 N5O 10 Na + [M+Na] + :834.4624,found:834.4620.

[0069] (24) Preparation of Compound 14

[0070] The synthesis method is the same as that of compound 1. The crude product was separated and purified by silica gel column chromatography (dichloromethane / ethyl acetate = 2 / 1-1 / 1) and preparative thin layer chromatography (ethyl acetate) to obtain white solid compound 14 (15 mg, 46%). 11H NMR (400 MHz, CDCl3) δ 7.27 (d, J = 2.1 Hz, 1H), 7.10 (d, J = 8.6 Hz, 2H), 6.92 (d, J = 8.9 Hz, 1H), 6.78 (d, J = 8.6 Hz, 2H), 6.08 (dd, J = 6.1, 1.7 Hz, 1H), 5.66 (dd, J = 10.0, 2.2 Hz, 1H), 5.40 (t, J = 8.4 Hz, 1H), 5.25 (dd, J = 6.5, 2.9 Hz, 1H), 5.02–4.91 (m, 2H), 4.80 (qt, J = 6.7, 1.8 Hz, 2H), 4.37 (s, 1H), 3.91–3.80 (m, 2H), 3.76 (s, 3H), 3.20 (dd, J = 14.8, 6.4 Hz, 1H), 3.06 (s, 3H), 2.97 (dd, J = 10.8, 3.9 Hz, 1H), 2.89 (s, 3H), 2.63 (m, 1H), 2.47 (m, 1H), 2.25 (m, 1H), 2.04 (d, J = 2.2 Hz, 1H), 1.99 (s, 3H), 1.46 (d, J = 6.7 Hz, 3H), 1.41 (m, 1H), 1.36 (m, 2H), 1.25 (m, 6H), 1.17 (d, J = 6.5 Hz, 3H), 0.98 (d, J = 6.5 Hz, 3H), 0.86 (d, J = 6.6 Hz, 6H), 0.82 (d, J = 6.2 Hz, 3H), 0.79 (d, J = 6.7 Hz, 3H); 13 13C NMR (100 MHz, CDCl3) δ 178.18, 174.85, 170.67, 170.00, 169.87, 169.04, 158.44, 154.28, 129.99, 129.00, 125.50, 113.98, 72.05, 68.62, 59.36, 58.76, 58.26, 57.12, 55.37, 52.08, 40.86, 37.36, 36.74, 33.77, 33.04, 31.64, 30.54, 30.48, 29.84, 29.16, 27.28, 23.14, 21.19, 19.65, 18.99, 18.59, 17.55, 17.07, 16.95, 14.26; HRMS (ESI) m / z: calcd for C 43 H 65 N5O 10 Na + [M + Na] + : 834.4624, found: 834.4620.

[0071] (25) Preparation of Compound 15

[0072] The synthesis method is the same as that of compound 1. The crude product was separated and purified by silica gel column chromatography (dichloromethane / ethyl acetate = 2 / 1-1 / 1) and preparative thin layer chromatography (ethyl acetate) to obtain white solid compound 15 (21 mg, 67%). 1 H NMR (400MHz, CDCl3) δ7.28(d,J=2.2Hz,1H),7.10(d,J=8.5Hz,2H),7.01(d,J=9.1Hz, 1H),6.78(d,J=8.6Hz,2H),6.08(dd,J=6.1,1.7Hz,1H),5.66(dd,J=10.0,2.1Hz,1H), 5.55(dd,J=10.4,6.1Hz,1H),5.25(dd,J=6.6,3.0Hz,1H),5.00(d,J=11.1Hz,1H),4. 96(dd,J=9.0,3.0Hz,1H),4.86–4.76(m,1H),4.37(s,1H),3.88–3.77(m,1H),3.75(s, 3H),3.19(dd,J=14.9,6.1Hz,1H),3.07(s,3H),2.95(dd,J=14.9,10.8Hz,1H),2.90( s,3H),2.63(m,1H),2.47(m,1H),2.24(m,1H),2.04(s,1H),1.99(s,3H),1.46(d,J=6. 7Hz,3H),1.25(m,4H),1.17(d,J=6.5Hz,3H),1.14(m,2H),1.05(d,J=6.7Hz,4H),0.99 (d,J=6.5Hz,3H),0.86(t,J=7.0Hz,4H),0.80(d,J=6.6Hz,3H),0.70(d,J=5.8Hz,3H); 13C NMR (100MHz, CDCl3) δ178.37,174.82,170.73,169.99,169.88,169.76,169.06,158.41,1 54.29,129.89,128.77,125.48,113.95,72.03,68.65,59.31,58.74,58.24,57.14,56.66 ,55.23,52.07,41.17,36.81,36.72,33.73,32.75,30.95,30.52,30.35,29.82,28.90,27 .28,23.16,21.18,19.73,18.96,18.61,17.56,17.27,17.06,14.21; HRMS(ESI)m / z:calcd for C 43 H 65 N5O 10 Na[M+Na] + :834.4624,found:834.4623.

[0073] (26) Preparation of Compound 16

[0074] The synthesis method is the same as that of compound 1. The crude product was separated and purified by silica gel column chromatography (dichloromethane / ethyl acetate = 2 / 1-1 / 1) and preparative thin layer chromatography (ethyl acetate) to obtain white solid compound 16 (11 mg, 34%). 1H NMR(400MHz,CDCl3)δ7.27(d,J=2.5Hz,1H),7.10(d,J=8.5Hz,2H),6.88(d,J=8.8Hz,1H),6.78(d,J=8.5Hz,2H),6.08(dd,J=6.1,1.7Hz,1H),5.66(dd,J=10.1,2.1Hz,1H),5.44(s,1H),5.26(dd,J=6.5,2.8Hz,1H),5.02–4.93(m,2H),4.85–4.76(m,1H),4.36(s,1H),3.89–3.78(m,2H),3.75(s,3H),3.21(dd,J=14.8,6.2Hz,1H),3.07(s,3H),3.03–2.92(m,1H),2.90(s,3H),2.69(m,1H),2.47(m,1H),2.24(m,1H),2.03(d,J=5.4Hz,1H),1.98(s,3H),1.77–1.66(m,1H),1.46(d,J=6.7Hz,3H),1.27(d,J=5.5Hz,1H),1.28–1.20(m,6H),1.17(d,J=6.5Hz,3H),1.09–1.00(m,1H),0.98(d,J=6.5Hz,3H),0.90–0.84(m,3H),0.81(d,J=6.5Hz,3H),0.79(d,J=6.7Hz,6H); 13 C NMR(100MHz,CDCl3)δ177.88,174.82,170.70,170.01,169.86,169.03,158.43,154.29,129.98,128.97,125.48,113.97,72.03,68.59,59.36,58.75,58.26,57.10,55.38,52.07,41.30,36.99,36.74,33.55,33.13,32.06,31.63,30.53,29.83,29.49,29.41,29.15,27.27,23.19,21.13,20.06,18.97,18.60,18.49,17.87,17.55,17.06,14.29;HRMS(ESI)m / z:calcd for C 43 H 65 N5O 10 Na + [M+Na] + :834.4624,found:834.4619.

[0075] (27) Preparation of Compound 17

[0076] The synthesis method is the same as that of compound 1. The crude product was separated and purified by silica gel column chromatography (dichloromethane / ethyl acetate = 2 / 1-1 / 1) and preparative thin layer chromatography (ethyl acetate) to obtain white solid compound 17 (13 mg, 41%). 1 H NMR (400MHz, CDCl3) δ7.27(d,J=2.0Hz,1H),7.10(d,J=8.6Hz,2H),7.02(d,J=9 .0Hz,1H),6.78(d,J=8.6Hz,2H),6.08(dd,J=6.0,1.6Hz,1H),5.66(dd,J=10.0 ,2.1Hz,1H),5.54(dd,J=10.7,6.0Hz,1H),5.25(dd,J=6.5,3.0Hz,1H),5.00(d ,J=11.1Hz,1H),4.96(dd,J=9.7,3.8Hz,1H),4.81(m,1H),4.37(s,1H),3.89–3 .78(m,2H),3.75(s,3H),3.22–3.10(m,1H),3.09(s,3H),3.02–2.92(m,1H),2. 90(s,3H),2.60–2.41(m,2H),2.24(m,1H),2.04(m,1H),1.98(s,3H),1.66–1.5 8(m,1H),1.46(d,J=6.8Hz,3H),1.25(m,8H),1.16(d,J=6.5Hz,3H),1.07(d,J= 6.7Hz, 3H), 0.99 (d, J = 6.5Hz, 3H), 0.86 (t, J = 7.2Hz, 4H), 0.80 (d, J = 6.7Hz, 3H); 13C NMR (100MHz, CDCl3) δ178.13,174.81,170.77,170.01,169.91,169.82,169.07,158.42,15 4.29,129.87,128.86,125.49,113.98,72.04,68.64,59.36,58.76,58.26,57.14,56.74,55 .24,52.08,36.74,36.22,34.17,32.66,31.73,31.57,31.07,30.55,30.32,29.83,29.44,2 7.30,27.25,22.82,21.17,18.97,18.62,17.64,17.56,17.07,14.19; HRMS(ESI)m / z:calcd for C 42 H 63 N5O 10 Na + [M+Na] + :820.4467,found:820.4462.

[0077] (28) Preparation of Compound 18

[0078] The synthesis method is the same as that of compound 1. The crude product was separated and purified by silica gel column chromatography (dichloromethane / ethyl acetate = 2 / 1-1 / 1) and preparative thin layer chromatography (ethyl acetate) to obtain white solid compound 18 (16.7 mg, 53%).

[0079] 1H NMR(400MHz,CDCl3)δ7.27(d,J=2.3Hz,1H),7.10(d,J=8.3Hz,2H),7.04(d,J=8.9Hz,1H),6.77(d,J=8.6Hz,2H),6.08(dd,J=6.1,1.7Hz,1H),5.65(dd,J=10.0,2.2Hz,1H),5.54(dd,J=10.7,5.9Hz,1H),5.25(dd,J=6.5,2.9Hz,1H),4.99(d,J=11.6Hz,1H),4.98–4.90(m,1H),4.85–4.75(m,1H),4.37(s,1H),3.89–3.77(m,2H),3.74(s,3H),3.22–3.10(m,1H),3.07(s,3H),3.02–2.91(m,1H),2.90(s,3H),2.58–2.41(m,2H),2.23(m,1H),2.03(m,J=2.8Hz,1H),1.98(s,3H),1.66–1.56(m,1H),1.46(d,J=6.7Hz,3H),1.24(m,8H),1.16(d,J=6.5Hz,4H),1.07(d,J=6.7Hz,3H),0.98(d,J=6.5Hz,3H),0.86(t,J=7.2Hz,4H),0.79(d,J=6.6Hz,3H); 13 C NMR(100MHz,CDCl3)δ178.13,174.82,170.77,170.00,169.91,169.83,169.07,158.43,154.29,129.86,128.86,125.49,113.98,72.04,72.00,68.64,59.36,58.76,58.26,57.14,56.74,55.24,52.09,36.74,36.22,34.17,32.66,32.06,31.73,31.57,31.07,30.55,30.32,29.83,29.79,29.74,29.49,29.43,27.30,27.25,22.83,22.80,21.16,18.97,18.61,17.64,17.56,17.07,14.26,14.19;HRMS(ESI)m / z:calcd for C 42 H 63 N5O 10 Na + [M+Na] +:820.4467,found:820.4466.

[0080] Example 2: Bioactivity of Majusculamide D derivatives against human pancreatic cancer cell line Panc-1, human lung adenocarcinoma cell line A549, human glioma cell line U251, human papillary lung adenocarcinoma cell line H820, human cervical cancer cell line HeLa, human astrocytic glioblastoma cell line U87, human hepatoma cell line SMMC-7721 and human colon cancer cell line HCT116

[0081] The cells to be tested were divided into 2×10 5 / mL cell suspension was added into a 96-well round-bottom cell culture plate, and the compounds to be tested were added respectively, with 3 wells for each test concentration. The plates were cultured at 37°C and 5% CO2 saturated humidity for 72 hours, and the absorbance (A) was measured at a wavelength of 570nm by MTT method using an enzyme-linked detector, and the inhibitory effect of the compounds of the present invention on the test cancer cells was calculated.

[0082] Table 1. Inhibitory activity of Majusculamide D derivatives against various cancer cells (IC 50 , nM)

[0083]

[0084] As shown in Table 1, the tested compounds showed strong anticancer activities against the tested cancer cell lines.

[0085] The above is an exemplary description of the implementation of the technical solution of the present disclosure. It should be understood that the protection scope of the present disclosure is not limited to the above-mentioned implementation. Any modification, equivalent substitution, improvement, etc. made by those skilled in the art within the spirit and principle of the present disclosure shall be included in the protection scope of the claims of this application.

Claims

1. A Majusculamide D derivative compound represented by formula (I) or a pharmaceutically acceptable salt thereof, In formula (I), R1 is hydrogen, methyl (R configuration) or methyl (S configuration); R2 is methyl (R configuration) or methyl (S configuration).

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The compound has the structure shown below:

3. A method for preparing the compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, characterized in that The compound represented by formula (I) is prepared using the compound represented by formula (II) as a raw material, 4. Use of the compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating cancer or an adjuvant drug for treating cancer, preferably, the cancer is selected from pancreatic cancer, lung adenocarcinoma, glioma, papillary lung adenocarcinoma, cervical cancer, human brain astrocytoma, liver cancer and colon cancer.

5. A pharmaceutical composition for treating cancer, comprising a therapeutically effective amount of the compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, and / or a pharmaceutically acceptable carrier and / or a combination of other anticancer drugs.

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