Majusculamide d derivatives, pharmaceutical compositions, and methods of making and using the same

Abstract

The application provides a Majusculamide D derivative or a pharmaceutically acceptable salt thereof as shown in formula (I), which has a good inhibiting effect on cancer cells and can be used for treating cancer.

Description

Technical Field

[0001] This invention provides a Majusculamide D derivative, and relates to a method for preparing the Majusculamide D derivative, and its use in the preparation of cancer treatment drugs. This invention belongs to the field of pharmaceutical technology. Background Technology

[0002] Majusculamide D was extracted and isolated from the marine cyanobacterium *Lyngbya majuscula* in the Marshall Islands as a secondary metabolite. Majusculamide D is a natural ester peptide product with a chiral fatty side chain, multiple natural amino acids, and an α,β-unsaturated ketone structure.

[0003] In 2019, Gerwick et al. completed the first total synthesis of Majusculamide D and found that it had good inhibitory activity against Panc-1 cells (IC50). 50 =0.32 nM), showing potential for further research and development. However, due to the low abundance of Majusculamide D in nature, its medicinal chemistry and mechanism have not been reported. This invention provides the synthesis of a fatty side-chain derivative of Majusculamide D, and the derivative has a therapeutic effect on cancer, exhibiting IC50 activity against various cancer cells. 50 The value is between 0.7 and 328.1 nM. Summary of the Invention

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

[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 following structure:

[0008]

[0009] A method for preparing the Majusculamide D derivative of formula (I) includes preparing the compound of formula (I) from a compound of formula (II).

[0010]

[0011] R1 and R2 are defined independently of each other as described above.

[0012] Use of a Majusculamide D derivative as shown in formula (I) in the preparation of a treatment for cancer or an adjunct to the treatment of cancer, wherein the cancer is selected from pancreatic cancer, lung adenocarcinoma, glioma, papillary lung adenocarcinoma, cervical cancer, human brain astrocytoma, liver cancer and colon cancer.

[0013] A pharmaceutical composition for treating cancer, comprising an effective amount of a pharmaceutically acceptable carrier of the Majusculamide D derivative of formula (I) or a combination with other anticancer drugs.

[0014] Beneficial effects

[0015] This invention provides a Majusculamide D derivative as shown in formula (I), which has a good inhibitory effect on a variety of cancer cells and can be used to treat cancer. Detailed Implementation

[0016] To help understand the present invention, the following embodiments are provided to further illustrate the invention, but these are not intended to limit the scope of protection of the invention.

[0017] Example 1: Synthesis of Majusculamide D derivatives

[0018]

[0019] (1) Preparation of compound 4

[0020] Compound 2 (5.53 g, 17.2 mmol) was added to a 250 mL round-bottom flask at room temperature and dissolved in THF (20 mL). The mixture was cooled to 0 °C, and a dioxane hydrochloride solution (4 N, 21.5 mL, 86.0 mmol) was added to the reaction system. The reaction was brought back to room temperature and stirred for 1 h. After the reaction was confirmed to be complete by TLC, the reaction solution was concentrated, and the high-boiling-point dioxane was removed by evaporation using an oil pump to obtain a white solid, which was directly used in the next step of the reaction.

[0021] Compound 3 (3.96 g, 18.07 mmol) and HATU (9.81 g, 25.8 mmol) were added to the reaction system at room temperature and dissolved in DCM (50 mL). The mixture was cooled to 0 °C, and DIPEA (8.5 mL, 51.6 mmol) was slowly added dropwise. The reaction was then brought to room temperature and stirred overnight. After TLC detection of complete reaction, the reaction solution was concentrated, diluted with ethyl acetate (100 mL), and the organic phase was first washed three times with 5% sodium bisulfate solution (3 × 100 mL), then washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 25 / 1-5 / 1) to give a colorless oily compound 4 (5.45 g, 75%), which then precipitated as a white solid.

[0022] 1 H NMR (400MHz, CDCl3) δ7.31(s,5H),5.46(d,J=9.3Hz,1H),5.18(d,J=12.2Hz,1H),5.07(d,J=12.2Hz,1H),4.88(d,J=10.5Hz,1H),4.43(dd,J=20.6,9.8 Hz,1H),3.95(d,J=6.9Hz,1H),3.74(s,1H),3.00(s,3H),2.21(m,1H),1.40 (s,9H),1.11(d,J=6.2Hz,3H),0.97(d,J=6.7Hz,3H),0.81(d,J=6.8Hz,3H). 13 C NMR (100MHz, CDCl3) δ173.56,170.31,156.17,135.46,128.66,128.58,128.54,128.51,128.47,128.41,128.36,128. 05,79.98,67.29,66.69,61.74,53.69,31.55,28.31,28.22,28.17,27.29,19.80,18.66,18.62.HRMS(ESI)m / z:calcd for C 22 H 34 N2O6Na + [M+Na] + :445.2309,found:445.2304.

[0023] (2) Preparation of compound 5

[0024] At room temperature, EA (60 mL) was added to a pre-dehydrated 250 mL Schlenk flask. The solvent was frozen with liquid nitrogen and a Dewar flask, then the air in the reaction system was removed by an oil pump, and then the system was brought to room temperature to purge with argon. This process was repeated three times. Compound 4 (5.42 g, 12.8 mmol), AgOTf (9.89 g, 38.5 mmol), Selectfluor (6.82 g, 19.2 mmol), and KF (2.98 g, 51.3 mmol) were rapidly added to the reaction flask, followed by the dropwise addition of CF3TMS (5.7 mL, 38.5 mmol) and 2-fluoropyridine (3.3 mL, 38.5 mmol). The mixture was stirred overnight at room temperature. After the reaction was confirmed to be complete by TLC, the reaction solution was filtered through a diatomaceous earth filter and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 30 / 1-5 / 1) to give a colorless oily compound 5 (3.26 g, 52%). 1 H NMR (400MHz, CDCl3) δ7.36–7.29(m,5H),5.28(d,J=8.5Hz,1H),5.17(d,J=12.1Hz,1H),5.08(d,J=12.1Hz,1H),4.92(d,J=10.5Hz,1H),4. 76–4.69(m,1H),4.46(m,1H),3.03(s,3H),2.25(m,1H),1.42(s,9H),1.20(d,J=6.4Hz,3H),1.01(d,J=6.6Hz,3H),0.83(d,J=6.8Hz,3H). 13 C NMR (100MHz, CDCl3) δ170.31,170.09,155.52,135.44,128.70,128.63,128.27,122.93,12 0.40,80.45,75.56,66.98,62.07,53.72,31.80,28.32,28.11,27.30,19.95,18.71,17.22. 19 F NMR(376MHz, CDCl3)δ-58.16,-58.50(rotamer).HRMS(ESI)m / z:calcd for C 23 H 33 N2O6F3Na + [M+Na] + :513.2183,found:513.2183.

[0025] (3) Preparation of compound 7

[0026] Compound 5 (3.23 g, 6.58 mmol) was added to a 250 mL round-bottom flask at room temperature and dissolved in THF (20 mL). The mixture was cooled to 0 °C, and a dioxane hydrochloride solution (4 N, 32.9 mL, 131.7 mmol) was added to the reaction system. The reaction was brought back to room temperature and stirred for 1 h. After the reaction was confirmed to be complete by TLC, the reaction solution was concentrated, and the high-boiling-point dioxane was removed by evaporation using an oil pump to obtain a white solid, which was directly used in the next step of the reaction.

[0027] Compound 6 (1.96 g, 6.58 mmol) and HATU (3.76 g, 9.88 mmol) were added to the reaction system at room temperature and dissolved in DCM (20 mL). The mixture was cooled to 0 °C, and DIPEA (3.3 mL, 19.75 mmol) was slowly added dropwise. The reaction was then brought to room temperature and stirred overnight. After TLC detection of complete reaction, the reaction solution was concentrated, diluted with ethyl acetate (100 mL), and the organic phase was first washed three times with 5% sodium bisulfate solution (3 × 100 mL), then washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 20 / 1-2 / 1) to give a pale yellow oily compound 7 (3.31 g, 75%).

[0028] 1 H NMR(400MHz, CDCl3)δ7.30(d,J=2.7Hz,5H),7.13(m,2H),7.00–6.65(m,3H),5.20–5.02(m ,2H),5.01(d,J=7.9Hz,1H),4.83(dd,J=29.8,9.3Hz,2H),4.50(p,J=6.4Hz,1H),3.27(dd, J=14.4,6.4Hz,1H),3.02(s,1H),2.97–2.82(m,3H),2.71(d,J=25.0Hz,3H),2.27–2.13(m ,1H),1.35(d,J=18.2Hz,9H),1.18(d,J=6.3Hz,3H),1.02–0.93(m,3H),0.79–0.67(m,3H); 13C NMR (100MHz, CDCl3) δ170.2,169.3,163.0,160.5,156.2,135.4,133.2,130.7,13 0.6,130.5,128.7,128.7,128.6,128.6,128.5,128.2,122.9,120.3,115.3,115. 1,80.8,75.2,67.1,66.9,61.9,59.8,52.3,52.2,33.3,32.0,31.6,31.0,29.8,29.7,29.4,28.3,28.2,28.1,28.1,27.3,22.8,19.8,19.7,18.7,18.6,17.3,14.2; 19 F NMR(376MHz, CDCl3)δ-57.9,-58.3,-58.4,-116.3,-116.6(rotamer); HRMS(ESI)m / z:calcd for C 33 H 43 N3O7F4Na + [M+Na] + :692.2929,found:692.2923.

[0029] (4) Preparation of compound 8

[0030] Compound 7 (4.70 g, 7.02 mmol) was added to a dry 250 mL three-necked round-bottom flask, dissolved in methanol (30 mL), and then Pd / C (10% wt, 470 mg) was added. The reaction system was first purged with argon three times, then with hydrogen three times, and stirred overnight under a hydrogen atmosphere at 1 atm. After the reaction was confirmed to be complete by TLC, the reaction solution was filtered through a diatomaceous earth filter (the Pd / C on the diatomaceous earth should not be dried too much to prevent ignition upon heating), washed with methanol, and then concentrated. The crude product was purified by silica gel column chromatography (dichloromethane / methanol = 100 / 1–20 / 1) to give a white, foamy solid compound 8 (3.50 g, 86%).

[0031] 1H NMR(400MHz, CDCl3)δ7.60(s,1H),7.18–7.07(m,3H),6.98–6.83(m,2H),5.18–5. 05(m,1H),4.90–4.72(m,2H),4.57(t,J=6.5Hz,1H),3.25(dd,J=14.4,7.0Hz,1H), 3.06(d,J=46.7Hz,3H),2.89(d,J=5.5Hz,1H),2.74(d,J=21.3Hz,3H),2.31–2.12 (m,1H),1.33(d,J=23.8Hz,12H),1.09–0.98(m,3H),0.76(dd,J=33.2,6.1Hz,3H); 13 C NMR (100MHz, CDCl3) δ173.6,172.1,170.7,170.4,169.6,169.3,163.0,160.6 ,156.6,156.3,133.0,130.8,130.7,130.5,125.4,122.9,120.4,115.6,115.4 ,115.2,81.3,81.1,75.2,65.0,62.4,61.1,59.7,59.6,52.4,33.5,33.1,32.3,31.0,30.4,29.8,29.6,28.2,28.2,27.5,27.3,19.9,19.7,18.8,18.2,17.5; 19 F NMR(376MHz, CDCl3)δ-57.9,-58.2,-116.3,-116.6.(rotamer); HRMS(ESI)m / z:calcd forC 26 H 37 N3O7F4Na + [M+Na] + :602.2460,found:602.2448.

[0032] (5) Preparation of compound 10

[0033] Compound 9 (8.3 g, 19.53 mmol) was added to a 250 mL round-bottom flask at room temperature and dissolved in THF (40 mL). The mixture was cooled to 0 °C, and a dioxane hydrochloride solution (4 N, 97.7 mL, 390.7 mmol) was added to the reaction system. The reaction was brought back to room temperature and stirred for 30 min. After the reaction was confirmed to be complete by TLC, the reaction solution was concentrated, and the high-boiling-point dioxane was removed by evaporation using an oil pump to obtain a white solid, which was directly used in the next step of the reaction.

[0034] Compound 8 (10.19 g, 17.58 mmol) and HATU (11.14 g, 29.30 mmol) were added to the reaction system at room temperature and dissolved in DCM (50 mL). The mixture was cooled to 0 °C, and DIPEA (9.7 mL, 58.60 mmol) was slowly added dropwise. The reaction was then brought to room temperature and stirred overnight. After TLC detection of complete reaction, the reaction solution was concentrated, diluted with ethyl acetate (100 mL), and the organic phase was first washed three times with 5% sodium bisulfate solution (3 × 100 mL), then washed with saturated brine (150 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel column chromatography (dichloromethane / ethyl acetate = 3 / 1-1 / 2) to give a white, foamy solid compound 10 (7.42 g, 55%).

[0035] 1 H NMR (400MHz, CDCl3) δ7.28(d,J=2.0Hz,1H),7.17–7.12(m,2H),7.02–6.80(m,3H),6.08(dd,J=6.0,1.6Hz,1H),5.59(dd,J=10.1,2. 0Hz,1H),5.10–4.99(m,2H),4.85–4.71(m,2H),4.57(q,J=6.0Hz,1H),4.37(s,1H),3.92(d,J=11.6Hz,1H),3.82(dd,J=11.6,4.4Hz ,1H),3.29(dd,J=14.4,6.4Hz,1H),3.12–2.90(m,4H),2.75(d,J=39.8Hz,4H),2.39(ddd,J=14.6,10.1,4.7Hz,1H),2.24(s,1H),2. 02(d,J=13.4Hz,1H),1.45(d,J=6.7Hz,3H),1.37(d,J=20.7Hz,9H),1.32(d,J=2.7Hz,3H),0.99(d,J=6.3Hz,3H),0.85–0.69(m,3H); 13C NMR (100MHz, CDCl3) δ174.6,170.4,170.0,169.0,163.0,160.6,156.2,154.4,133.3,130.7,130.7,125.4,122.9,120.3,115.4,115. 2,80.9,74.9,71.9,60.3,59.5,58.6,58.3,57.2,52.5,52.2,38.8,36.7,33.4,31.6,30.6,29.8,28.3,27.4,18.9,18.4,17.9,17.0; 19 F NMR(376MHz, CDCl3)δ-58.3,-58.5,-116.3,-116.6(rotamer); HRMS(ESI)m / z:calcd for C 36 H 49 N5O9F4Na + [M+Na] + :794.3359,found:794.3352.

[0036] (6) Preparation of compound 2-1

[0037] Compound 10 (7.42 g, 9.61 mmol) was added to a 250 mL round-bottom flask at room temperature and dissolved in 30 mL of THF. The mixture was cooled to 0 °C, and a dioxane hydrochloride solution (4 N, 96.1 mL, 384.6 mmol) was added to the reaction system. The reaction was brought back to room temperature and stirred for 2 h. After the reaction was confirmed to be complete by TLC, the reaction solution was concentrated, and the high-boiling-point dioxane was removed by evaporation using an oil pump to obtain a white solid, which was directly used in the next step of the reaction.

[0038] Compound 11 (4.56 g, 28.84 mmol) and HATU (10.97 g, 28.84 mmol) were added to the reaction system at room temperature and dissolved in DCM (30 mL). The mixture was cooled to 0 °C, and DIPEA (9.53 mL, 57.68 mmol) was slowly added dropwise. The reaction was brought back to room temperature and stirred overnight. After TLC detection of complete reaction, the reaction solution was concentrated, diluted with ethyl acetate (100 mL), and the organic phase was first washed three times with 5% sodium bisulfate solution (3 × 100 mL), then washed with saturated brine (150 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel column chromatography (dichloromethane / methanol = 100 / 1-20 / 1), and lyophilized to give compound 2-1 (4.83 g, 62%) as a white solid powder.

[0039] 1H NMR(400MHz,CDCl3)δ7.28(d,J=2.0Hz,1H),7.20–7.10(m,2H),6.97–6.87(m,3H),6.07(dd,J=6.1,1.6Hz,1H),5.58(dd,J=10.1,2.1Hz,1H),5.46(dd,J=9.8,6.6Hz,1H),5.09–4.98(m,2H),4.79(qt,J=6.7,1.8Hz,1H),4.53(p,J=6.2Hz,1H),4.36(d,J=4.7Hz,1H),3.91(d,J=11.9Hz,1H),3.84(dd,J=11.6,4.4Hz,1H),3.22(dd,J=14.8,6.5Hz,1H),3.00(s,3H),2.95(d,J=5.1Hz,1H),2.88(s,3H),2.53(h,J=6.8Hz,1H),2.39(ddd,J=14.6,10.1,4.8Hz,1H),2.22(dq,J=10.9,6.6Hz,1H),2.01(dd,J=14.5,1.9Hz,1H),1.45(d,J=6.8Hz,3H),1.42–1.36(m,1H),1.30(d,J=6.2Hz,3H),1.25–1.09(m,8H),1.04(d,J=6.7Hz,3H),0.99(d,J=6.5Hz,3H),0.97–0.90(m,2H),0.86(t,J=7.2Hz,3H),0.75(d,J=6.7Hz,3H); 13 C NMR(100MHz,CDCl3)δ178.2,174.4,170.1,169.9,169.0,168.9,168.8,168.8,162.9,160.5,154.3,132.5,132.5,130.5,130.4,125.3,122.7,120.2,115.4,115.2,74.8,74.8,71.7,59.3,58.5,58.2,57.0,56.9,52.2,36.6,36.1,34.0,32.8,31.6,30.9,30.5,29.2,27.3,27.1,22.6,18.8,18.4,17.9,16.9,14.0; 19 F NMR(376MHz,CDCl3)δ-58.3,-116.2;HRMS(ESI)m / z:calcdfor C 40 H 57 N5O8F4Na + [M+Na]+ :834.4035,found:834.4029.

[0040] Example 2: Bioactivity of Majusculamide D derivative against human pancreatic cancer cell line Panc-1

[0041] Prepare 2×10 cells for testing. 5 / mL of cell suspension was added to 96-well round-bottom cell culture plates, and the test compound was added to each well, with 3 wells for each test concentration. The plates were incubated at 37°C and 5% CO2 saturated humidity for 72 hours. The absorbance (A) was measured at 570nm using the MTT assay, and the inhibitory effect of the compound on the test cancer cells was calculated. The inhibitory activity of the Majusculamide D derivative (compound 1) against the human pancreatic cancer cell line Panc-1 was IC50. 50 =3.83±1.54nM.

[0042] The above description provides an exemplary account of the implementation methods of the technical solution disclosed herein. It should be understood that the scope of protection of this disclosure is not limited to the above-described embodiments. Any modifications, equivalent substitutions, or improvements made by those skilled in the art within the spirit and principles of this disclosure should be included within the scope of protection of the claims of this application.

Claims

1. A Majusculamide D derivative compound as shown in 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 following structure:

3. A method for preparing the compound of claim 1 or 2 or a pharmaceutically acceptable salt thereof, characterized in that... Compound of formula (I) was prepared from compound of formula (II). R1 and R2 are independently defined as described in claim 1 or 2.

4. Use of the compound of claim 1 or 2 or a pharmaceutically acceptable salt thereof in the preparation of a treatment for cancer or an adjunct to the treatment of cancer, preferably, the cancer being selected from pancreatic cancer, lung adenocarcinoma, glioma, papillary lung adenocarcinoma, cervical cancer, human astrocytoma, liver cancer, and colon cancer.

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

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