Indolocarbazole glycoalkaloid, preparation method thereof and application of indolocarbazole glycoalkaloid in anti-leukemia drugs

Abstract

The invention belongs to the field of organic synthesis and medicinal chemistry, and particularly relates to indolocarbazole glycoalkaloids, a preparation method thereof and application of the indolocarbazole glycoalkaloids in anti-leukemia drugs. The structure of the indolocarbazole glycoalkaloid or the pharmaceutically acceptable salt of the indolocarbazole glycoalkaloid is as shown in formulas 1-4: # imgabs0 #. Pharmacological tests prove that the indolocarbazole glycoalkaloids as shown in the formulas 1-4 have anti-leukemia activity and can be used for preparing drugs for resisting myeloid leukemia, myeloid monocytic leukemia and lymphocytic leukemia.

Description

Technical Field

[0001] The invention relates to the fields of organic synthesis and medicinal chemistry, and in particular to a class of indolecarbazole glycoside alkaloids with novel structures, a pharmaceutical composition containing the same, a preparation method thereof, and an application of the same in anti-leukemia drugs. Background Art

[0002] Leukemia is a malignant tumor of the blood system, and its main cause is the obstruction of hematopoietic stem cell differentiation, which leads to abnormal cell proliferation. In recent years, children have gradually become a high-risk group for leukemia, and its incidence rate in adults has also continued to rise. The prognosis of leukemia is generally poor, and the mortality rate is high, posing a serious threat to people's health and life. Currently, chemotherapy is the main treatment for leukemia, but its limited therapeutic effect and serious adverse reactions and drug resistance are still clinical problems that need to be solved urgently. It is crucial to develop novel structural drugs for the treatment of leukemia.

[0003] Due to the complex and unique chemical structure and diverse biological activities of indolecarbazole glycoside alkaloids, this type of compound has great application value and broad application prospects. Among this type of compound, two compounds have entered clinical trials and even been approved for marketing. Midostaurin is a multi-tyrosine kinase receptor inhibitor used to treat FMS-like tyrosine kinase 3 (FLT3)-positive acute myeloid leukemia (AML). Lestaurtinib is an orally bioavailable tyrosine kinase receptor inhibitor that has therapeutic effects on acute myeloid leukemia, chronic myeloid leukemia, and acute lymphocytic leukemia. Although midostaurin is a marketed drug for the treatment of FLT3-positive AML, it has adverse reactions such as febrile neutropenia, mucositis, and vomiting, which seriously hinders its application in clinical treatment. Lestaurtinib is currently in Phase III clinical trials for acute lymphocytic leukemia and has not been approved for marketing. Therefore, the development of new indolecarbazole glycoalkaloids that are easy to prepare and have less toxic side effects is of great significance for the treatment of AML. Summary of the invention

[0004] In view of the above problems, the purpose of the present invention is to provide a class of novel indolecarbazole glycoside alkaloids and a preparation method thereof, and at the same time provide the use of such compounds in anti-leukemia drugs.

[0005] The objective of the present invention is achieved through the following technical solutions:

[0006] Indolecarbazole glycoside alkaloid or a pharmaceutically acceptable salt thereof as shown in Formula 1-4:

[0007]

[0008] The method for preparing the indolecarbazole glycoside alkaloid as shown in formula 1-4 is characterized by comprising the following steps:

[0009]

[0010] in,

[0011] R 1 : is H, methyl (CH3) or benzyl (Bn);

[0012] R 2 : is H, hydroxyl (OH) or benzyloxy (BnO).

[0013] Step 1: Compounds 5, 6, a phosphine reagent and an azo reagent are dissolved in a first solvent and reacted at -78°C to room temperature for 6 to 24 hours; then, tert-butyloxycarbonyl (Boc) is removed under silica gel / 90°C; and tert-butyldiphenylsilyl (TBDPS) is removed under tetrabutylammonium fluoride (TBAF) conditions to obtain compound 7;

[0014] Step 2: dissolving compound 7 in a second solvent, adding the first catalyst, and reacting at 60° C. to 110° C. for 6 to 24 hours to obtain compound 8;

[0015] Step 3: Compound 8, triphenylphosphine, imidazole and iodine are dissolved in a third solvent, and reacted at 60° C. to 110° C. for 1 to 8 hours; then, the olefin is eliminated under 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) conditions to obtain compound 9;

[0016] Step 4: Compound 9, DBU, potassium iodide and iodine are dissolved in tetrahydrofuran (THF) and reacted at 0°C to room temperature for 10 to 60 minutes; then iodine is removed under the conditions of hydrogen, 20% Pd(OH)2 and N,N-diisopropylethylamine (DIPEA) to obtain compound 10;

[0017] Step 5: Dissolve compound 8 or 10 in a fourth solvent, add a second catalyst, and react at 0°C to room temperature for 12 to 24 hours under hydrogen conditions; then react at room temperature for 1 hour under ammonia conditions to obtain an indolecarbazole glycoside alkaloid represented by formula 1-4.

[0018] In step 1, the phosphine reagent is selected from any one of compounds P1 to P12, preferably: phosphine reagent P1; the azo reagent is selected from any one of compounds A1 to A5, preferably: azo reagent A1; the molar ratio of compounds 5, 6, phosphine reagent and azo reagent is: 1:3.0:6.0:6.0 to 1:4.0:12.0:12.0; the first solvent is: THF, dichloromethane (DCM), toluene (PhMe) or acetonitrile (MeCN), preferably: THF; the concentration of compound 5 in the first solvent is: 0.05 to 0.2 mmol / mL;

[0019] The structures of the phosphine reagent and the azo reagent are shown below:

[0020]

[0021] In step 2, the first catalyst is palladium acetate, palladium chloride or copper chloride, preferably palladium acetate; the molar ratio of compound 7 to the first catalyst is 1:1.0 to 1:2.0; the second solvent is acetic acid (AcOH), N,N-dimethylformamide (DMF) or acetone, preferably AcOH; the concentration of compound 7 in the second solvent is 0.01 to 0.1 mmol / mL.

[0022] In step 3, the third solvent is THF or PhMe, preferably THF; the concentration of compound 8 in the third solvent is 0.01-0.1 mmol / mL.

[0023] In step 5, the second catalyst is: 10% palladium carbon, 20% palladium hydroxide carbon or 10% palladium carbon / 20% palladium hydroxide carbon, preferably 10% palladium carbon; the fourth solvent is THF / methanol (MeOH), THF / isopropanol (i-PrOH) or THF / i-PrOH / H2O, preferably THF / i-PrOH / H2O; the concentration of compound 8 or 10 in the fourth solvent is 0.01-0.1 mmol / mL.

[0024] The pharmaceutically acceptable salt of the indolecarbazole glycoside alkaloid of the present invention is a salt formed by a pharmaceutically acceptable inorganic acid or organic acid, preferably hydrochloric acid, sulfuric acid, phosphoric acid, hydrobromic acid, maleic acid, fumaric acid, citric acid, methanesulfonic acid, p-toluenesulfonic acid, trifluoroacetic acid, tartaric acid or acetic acid.

[0025] Pharmacological experiments have shown that the indolecarbazole glycoside alkaloids shown in Formula 1-4 have anti-leukemia activity and can be used to prepare anti-leukemia drugs.

[0026] The anti-leukemia drugs are respectively anti-myeloid cell leukemia, anti-myelomonocytic leukemia and anti-lymphocytic leukemia drugs.

[0027] The anti-leukemia activity test uses anti-human myeloid leukemia cells HL-60, anti-human myelomonocytic leukemia cells MV-4-11, and anti-human T lymphocyte leukemia cells Jurkat (Clone E6-1).

[0028] The indolecarbazole glycoside alkaloids provided by the present invention have significant anti-leukemia activity. Compounds 1 to 4 all have good anti-leukemia activity against human myeloid leukemia cells HL-60, IC 50 0.69~1.91μM. For human chronic myeloid leukemia cells K-562, compound 2 (IC 50 =1.68 μM) and 4 (IC 50 =1.90 μM) showed similar antitumor activity to midostaurin (IC 50 =1.78 μM) and compound 1 (IC 50 =1.05 μM) and 3 (IC 50 =0.49 μM) had stronger antitumor activity than midostaurin. Compounds 1 to 4 all showed significant antitumor activity against human myelomonocytic leukemia cells MV-4-11 (IC 50 =5.6~10.3nM), and the activity was better than midostaurin (IC 50 =11.3 nM). Antitumor activity of compounds 1 to 4 against human T lymphocyte leukemia Jurkat cells (Clone E6-1) (IC 50 =0.22~0.85μM) are stronger than midostaurin (IC 50 =0.96 μM).

[0029] The indolecarbazole glycoside alkaloid compound prepared by the present invention has significant anti-leukemia activity and a clear mechanism of action. Further analysis shows that the mechanism of action is that the indolecarbazole glycoside alkaloid 1 significantly inhibits the expression of p-FLT3, p-STAT5, p-AKT and p-Erk in MV4-11 cells in a dose-dependent manner within the range of 6.25 to 100 nM, and inhibits the activity of STAT5 downstream transcription factors by downregulating the phosphorylation level of the FLT3 pathway, resulting in cell arrest in the G2 / M phase of the cell cycle, thereby inducing MV-4-11 cell apoptosis and exerting anti-leukemia activity. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 This is a graph showing the results of the apoptosis experiment of compound 1 on MV-4-11 cells.

[0031] A)-E) Typical flow cytometry results of groups A to E respectively.

[0032] Figure 2This is a graph showing the results of the apoptosis experiment of compound 1 on MV-4-11 cells.

[0033] A) is a bar graph of early apoptotic cells; B) is a bar graph of normal cells. All statistics are compared with group A, ns means no significant difference, **p<0.005, ****p<0.0001.

[0034] Figure 3 This is a graph showing the results of a cell cycle experiment of compound 1 on MV-4-11 cells.

[0035] Statistical data of MV-4-11 cells in group AE at each cycle. The statistical data of each period were compared with those in group A, ns means no significant difference, **p<0.01, ***p<0.001, ****p<0.0001.

[0036] Figure 4 Western blot analysis of the expression levels of p-FLT3, FLT3, p-STAT5, STAT5, p-AKT, AKT, p-Erk and Erk in MV-4-11 cells treated for 4 hours with different concentrations of compound 1. DETAILED DESCRIPTION

[0037] The present invention is further described in detail below by way of examples, but the present invention is not limited to the following examples.

[0038] Example 1: Preparation of Compound 1:

[0039] Step 1: Preparation of compound 7a:

[0040]

[0041] Under argon conditions, triphenylphosphine (2.8 g, 10.8 mmol, 6.0 equiv) and activated Molecular sieves (2.0 g) were added to a 50.0 mL reaction bottle, tetrahydrofuran (10.0 mL) was added to dissolve, diisopropyl azodicarboxylate (DIAD) (2.1 mL, 10.8 mmol, 6.0 equiv) in tetrahydrofuran (5.0 mL) was added at -78 ° C, stirred for 1 h, and tetrahydrofuran (5.0 mL) solutions of compound 5 (1.0 g, 1.8 mmol, 1.0 equiv) and compound 6a (3.7 g, 5.4 mmol, 3.0 equiv) were added at -78 ° C, and the temperature was naturally raised to room temperature for 12 h. After the reaction was completed, it was filtered, diluted with ethyl acetate, washed with saturated ammonium chloride solution and saturated sodium chloride solution in turn, dried over anhydrous sodium sulfate, filtered and concentrated, and purified by silica gel column chromatography (PE: EA = 8: 1) to obtain a red foam. Under argon, add red foam (1.2 g, 984.8 μmol, 1.0 equiv) to a 50.0 mL reaction bottle, add toluene (25.0 mL) to dissolve, add silica gel (2.0 g), and heat at 90 ° C for 12 h. After the reaction is completed, filter and concentrate, and purify by silica gel column chromatography (PE: EA = 4: 1) to obtain red foam. Under argon, add red foam (598.0 mg, 534.7 μmol, 1.0 equiv) to a 25.0 mL reaction bottle, add tetrahydrofuran (20.0 mL) to dissolve, add TBAF (1.6 mL, 1.6 mmol, 3.0 equiv, 1 M in THF) at 0 ° C, and react at room temperature for 30 min. After the reaction, the mixture was diluted with ethyl acetate, washed with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered and concentrated, and purified by silica gel column chromatography (PE:EA=2:1) ​​to obtain compound 7a (400.0 mg, 454.5 μmol, 85%) as red foam. 1 H NMR (400MHz, CDCl3) δ: 8.56 (d, J = 2.8Hz, 1H), 7.87 (s, 1H), 7.57 (d, J = 2.8Hz, 1H), 7.39 (d, J = 8.3Hz, 1H),7.34–7.11(m,16H),7.09–6.87(m,6H),6.83(d,J=8.1Hz,1H),6.74–6.62(m,3H),6.48(t,J=7.6 Hz,1H),5.42(d,J=8.9Hz,1H),5.13(s,2H),4.92–4.77(m,3H),4.69–4.60(m,3H),4.16(d,J=10.1H z,1H),3.92(t,J=8.9Hz,1H),3.86–3.75(m,2H),3.75–3.64(m,3H),3.61–3.50(m,1H),1.95(s,1H).13 C{ 1 H}NMR(101MHz,CDCl3)δ:171.8,171.7,138.3,137.8,137.7,136.8,136.0,135.8,129.5 ,128.7,128.63,128.56,128.5,128.4,128.3,128.24,128.18,128.1,128.0,127.8,127 .7,127.2,126.5,125.5,123.0,122.7,122.6,121.9,121.1,120.5,111.3,110.9,107.6 ,107.0,85.7,85.3,81.5,78.3,77.2,75.8,75.3,75.0,71.6,67.2,61.7.HRMS(ESI)m / z calcd for C 55 H 49 O8N3Na[M+Na + ]902.3412found 902.3425.

[0042] Step 2: Preparation of compound 8a:

[0043]

[0044] Under argon, compound 7a (400.0 mg, 454.5 μmol, 1.0 equiv) was added to a 50.0 mL reaction bottle, acetic acid (10.0 mL) was added to dissolve, palladium acetate (204.1 mg, 909.0 μmol, 2.0 equiv) was added, and the mixture was heated at 70°C for 8 h. After the reaction, the mixture was filtered, diluted with ethyl acetate, washed with saturated sodium bicarbonate solution, water and saturated sodium chloride solution in sequence, dried over anhydrous sodium sulfate, filtered and concentrated, and purified by silica gel column chromatography (PE: EA = 4: 1) to obtain compound 8a (308.0 mg, 350.8 μmol, 77%) as yellow-green foam. 1H NMR (400MHz, CDCl3) δ: 10.84 (s, 1H), 9.13 (d, J = 8.0Hz, 1H), 8.98 (d, J = 7.9Hz, 1H), 7.46–7.35 (m, 8H), 7. 33–7.22(m,4H),7.20–7.11(m,8H),7.04(t,J=7.4Hz,1H),6.86(t,J=7.4Hz,1H),6.79–6.68(m,2H),5.9 2–5.83(m,2H),5.73(d,J=8.9Hz,1H),5.00–4.89(m,3H),4.81(d,J=11.0Hz,1H),4.74(s,2H),4.62(s,2 H),4.25(d,J=11.1Hz,1H),4.21–4.06(m,2H),3.89–3.78(m,2H),3.74–3.59(m,2H),2.68–2.55(m,2H). 13 C{ 1 H}NMR(101MHz, CDCl3)δ:169.5,168.8,141.4,141.0,138.0,137.9,137.7,135.8,130.0 ,128.8,128.6,128.5,128.4,128.3,128.1,128.0,127.84,127.80,127.7,127.6,127.4 ,127.3,125.9,125.5,122.1,122.0,121.10,121.05,120.4,119.2,118.3,118.2,111.5 ,110.3,85.5,84.4,81.2,78.2,76.0,75.2,75.0,74.5,71.6,66.7,59.9.HRMS(ESI)m / z calcd for C 55 H 47 O8N3Na[M+Na + ]900.3255found 900.3271.

[0045] Step 3: Preparation of compound 9a:

[0046]

[0047] Under argon conditions, compound 8a (308.0 mg, 350.8 μmol, 1.0 equiv) was added to a 50.0 mL reaction bottle, and then tetrahydrofuran (10.0 mL) was added to dissolve, and triphenylphosphine (138.0 mg, 526.2 μmol, 1.5 equiv), imidazole (75.0 mg, 1.1 mmol, 3.0 equiv) and iodine (134.0 mg, 526.2 μmol, 1.5 equiv) were added, and heated at 80°C for 2 h. After the reaction was completed, the mixture was concentrated and purified by silica gel column chromatography (PE: EA = 6: 1) to obtain a yellow-green foam. Under argon, yellow-green foam (295.0 mg, 298.2 μmol, 1.0 equiv) was added to a 50.0 mL reaction bottle, tetrahydrofuran (10.0 mL) was added to dissolve, DBU (896.4 μL, 6.0 mmol, 20.0 equiv) was added, and the mixture was heated at 80°C for 1 h. After the reaction, the mixture was diluted with ethyl acetate, washed with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (PE: EA = 8: 1) to obtain compound 9a (172.0 mg, 199.8 μmol, 67%) as yellow-green foam. 1 H NMR(400MHz, CDCl3)δ:9.54(s,1H),9.37(d,J=7.9Hz,1H),9.30–9.22(m,1H),7.68–7.15(m, 20H),7.08–7.00(m,1H),6.89–6.79(m,2H),6.77–6.69(m,2H),6.37–6.27(m,2H),5.40(s,2H ),5.09–4.93(m,2H),4.81(s,3H),4.73–4.68(m,1H),4.66–4.52(m,2H),4.41(d,J=4.0Hz,1 H),4.28(d,J=8.7Hz,1H),4.20–4.05(m,1H),3.96(d,J=11.5Hz,1H),3.65(d,J=11.6Hz,1H). 13 C{ 1H}NMR(101MHz, CDCl3)δ:169.7,169.6,152.5,141.6,140.4,137.8,137.2,136. 8,135.7,129.5,128.80,128.75,128.39,128.36,128.3,128.2,128.1,127.88,1 27.85,127.7,127.6,127.5,127.4,125.9,125.5,122.2,121.8,121.1,120.7,1 11.0,109.9,97.5,82.4,79.4,79.2,75.0,72.7,71.5,70.6,66.9.HRMS(ESI)m / z calcd for C 55 H 45 O7N3Na[M+Na + ]882.3150 found 882.3155.

[0048] Step 4: Preparation of compound 10a:

[0049]

[0050] Under argon, compound 9a (172.0 mg, 199.8 μmol, 1.0 equiv) was added to a 25.0 mL reaction bottle, tetrahydrofuran (5.0 mL) was added to dissolve, DBU (60.0 μL, 399.6 μmol, 2.0 equiv) was added at 0°C, stirred for 10 min, potassium iodide (199.0 mg, 1.2 mmol, 6.0 equiv) and iodine (305.0 mg, 1.2 mmol, 6.0 equiv) were added, and the reaction was carried out at 15°C for 10 min. After the reaction, the mixture was diluted with ethyl acetate, washed with saturated sodium thiosulfate solution and saturated sodium chloride solution in sequence, dried over anhydrous sodium sulfate, filtered and concentrated, and purified by silica gel column chromatography (PE:EA=8:1) to obtain a yellow-green foam. The yellow-green foam (150.0 mg, 151.8 μmol, 1.0 equiv) was added to a 25.0 mL reaction bottle, tetrahydrofuran (2.0 mL) and methanol (2.0 mL) were added to dissolve, N, N-diisopropylethylamine (DIPEA) (79.0 μL, 455.4 μmol, 3.0 equiv) and 20% palladium hydroxide on carbon (10.0 mg) were added, and after hydrogen replacement, the mixture was stirred at room temperature for 5 h. After the reaction was completed, the mixture was filtered and concentrated, and purified by silica gel column chromatography (PE: EA = 8: 1) to obtain compound 10a (110.0 mg, 127.5 μmol, 85%) as a yellow-green foam. 1H NMR(400MHz, CDCl3)δ:9.23(d,J=7.6Hz,1H),9.10(d,J=7.9Hz,1H),7.49–6.99(m,22H),6.91–6.84(m,4H),6.28(d,J=3.1Hz,1H ),5.27(s,2H),4.81(d,J=11.5Hz,1H),4.70(s,2H),4.60–4.41(m,5H),4.17(d,J=11.2Hz,1H),4.14–4.04(m,2H),2.01(s,3H). 13 C{ 1 H}NMR(101MHz, CDCl3)δ:169.6,169.4,139.6,138.8,137.8,137.1,137.0,136.3,129 .7,129.1,128.7,128.5,128.44,128.40,128.35,128.2,127.9,127.81,127.6,127.4, 127.3,127.0,126.3,126.1,124.0,122.4,121.4,121.3,120.2,119.2,117.7,116.8,1 12.0,108.7,96.2,85.5,79.7,79.5,78.2,74.2,73.9,71.5,66.9,24.9.HRMS(ESI)m / z calcd for C 56 H 46 O9N3[M+HCO2 - ]904.3240 found 904.3233.

[0051] Step 5: Preparation of compound 1:

[0052]

[0053] Compound 10a (110.0 mg, 127.5 μmol, 1.0 equiv) was added to a 25.0 mL reaction bottle, and then tetrahydrofuran (3.0 mL) and isopropanol (3.0 mL) were added to dissolve, and 10% palladium carbon (50.0 mg) was added. After hydrogen replacement, the reaction was stirred at room temperature for 12 hours. After the reaction was completed, the mixture was filtered and concentrated; the concentrated crude product was dissolved in tetrahydrofuran (2.0 mL) and methanol (6.0 mL), and ammonia water (2.0 mL) was added. The reaction was stirred at room temperature for 1 hour. After the reaction was completed, the mixture was concentrated and purified by silica gel column chromatography (CH2Cl2: MeOH = 20: 1) to obtain compound 1 (38.0 mg, 80.3 μmol, 63%) as yellow foam. 1 H NMR(400MHz, DMSO-d6)δ:11.14(s,1H),9.18(d,J=7.9Hz,1H),9.00(d,J=8.0Hz,1H),7.90(d,J=8.5Hz,1H),7.83(d,J=8.3Hz,1H),7.67–7.59(m,2H ),7.50–7.37(m,2H),6.51(d,J=6.0Hz,1H),6.30(d,J=6.2Hz,1H),5.80(s ,1H),5.38(s,1H),4.39(t,J=5.7Hz,1H),3.90–3.74(m,2H),1.86(s,3H). 13 C{ 1 H}NMR(101MHz,DMSO-d6)δ:171.1,170.8,139.5,139.2,129.2,128.5,127.0,126.9,125.1,124.6,123.2,121 .2,120.9,120.8,120.7,120.0,116.1,115.3,113.1,110.3,98.2,86.0,76.3,73.5,71.6,24.4.HRMS(ESI)m / z calcd forC 26 H 20 O6N3[M+H + ]470.1347 found 470.1339.

[0054] Example 2: Preparation of Compound 2:

[0055] Step 1: Preparation of compound 7b:

[0056]

[0057] According to the synthesis method of compound 7a, compound 5 (500.0 mg, 913.1 μmol, 1.0 equiv) and compound 6b (2.1 g, 3.6 mmol, 4.0 equiv) were used as raw materials to obtain compound 7b (278.6 mg, 360.0 μmol, 39% for 3 steps) as a red foam. 1H NMR(400MHz, CDCl3)δ:8.74(s,1H),7.69(d,J=2.8Hz,1H),7.56(d,J=8.3Hz,1H),7.49(s,1H),7.44–7.17(m, 16H),7.13(t,J=7.7Hz,1H),6.97(t,J=7.6Hz,1H),6.93–6.82(m,2H),6.75(t,J=7.5Hz,1H),6.03(d,J=4.3H z,1H),5.28–5.17(m,2H),4.83(d,J=11.2Hz,1H),4.72(s,2H),4.65(d,J=11.2Hz,1H),4.50–4.38(m,2H),3. 80–3.62(m,3H),3.59(t,J=8.5Hz,1H),3.22–3.12(m,1H),2.54–2.44(m,1H),2.17–2.04(m,1H),1.75(s,1H). 13 C{ 1 H}NMR(101MHz,CDCl3)δ:171.8,171.6,138.2,138.1,137.8,136.6,136.1,129 .1,128.7,128.6,128.5,128.4,128.1,128.04,127.98,127.9,127.8,127.2,1 26.7,124.6,123.2,123.1,122.4,122.1,121.4,120.7,111.8,111.5,107.1,1 06.8,80.2,77.1,74.6,73.0,72.2,71.7,67.2,61.8,32.5,29.7.HRMS(ESI)m / z calcd for C 48 H 43 O7N3Na[M+Na + ]796.2993found 796.2993.

[0058] Step 2: Preparation of compound 8b:

[0059]

[0060] According to the synthesis method of compound 8a, compound 7b (373.6 mg, 0.48 mmol, 1.0 equiv) was used as a raw material to obtain compound 8b (215.3 mg, 0.28 mmol, 58%) as a yellow-green foam. 1H NMR (400MHz, CDCl3) δ: 10.97 (s, 1H), 9.13 (d, J = 7.4Hz, 1H), 8.98 (d, J = 7.9Hz, 1H), 7.48–7. 36(m,10H),7.32–7.16(m,11H),5.89(dd,J=11.1,2.6Hz,1H),5.08–5.01(m,3H),4.88(d,J =11.1Hz,1H),4.71(s,1H),4.53–4.43(m,2H),4.29–4.22(m,1H),4.17(d,J=11.3Hz,1H),4 .09(t,J=9.3Hz,1H),3.89–3.78(m,2H),2.63(s,1H),2.07–1.97(m,1H),1.97–1.84(m,1H). 13 C{ 1 H}NMR(101MHz, CDCl3)δ:169.5,169.0,140.9,139.7,138.2,137.69,137.65,129.8,12 8.74,128.67,128.6,128.5,128.41,128.37,128.3,128.0,127.9,127.8,127.7,127.6 ,127.1,127.0,126.0,125.3,122.5,121.9,121.3,120.7,120.4,119.2,118.2,117.9, 111.4,109.0,82.0,79.4,78.4,75.1,74.6,71.6,71.5,66.7,60.3,35.6.HRMS(ESI)m / z calcd for C 48 H 41 O7N3Na[M+Na + ]794.2837found 794.2844.

[0061] Step 3: Preparation of compound 9b:

[0062]

[0063] According to the synthesis method of compound 9a, compound 8b (50.0 mg, 64.8 μmol, 1.0 equiv) was used as a raw material to obtain compound 9b (23.0 mg, 30.5 μmol, 72% for 2 steps) as a yellow-green foam. 1H NMR(400MHz, CDCl3)δ:9.98(s,1H),9.34(dd,J=8.0,1.3Hz,1H),9.28–9.19(m,1H),7.55–7.4 5(m,5H),7.45–7.38(m,4H),7.36–7.26(m,10H),7.24–7.19(m,1H),6.98(dd,J=9.3,6.6Hz,1H ),6.94–6.86(m,1H),5.28(s,2H),5.03–4.96(m,2H),4.74(s,2H),4.71(d,J=11.6Hz,1H),4. 67(d,J=1.6Hz,1H),4.64(s,2H),4.38(d,J=4.1Hz,1H),4.24–4.16(m,1H),2.66–2.53(m,2H). 13 C{ 1 H}NMR(101MHz,CDCl3)δ:169.6,169.5,152.9,140.5,140.3,137.9,137.3,13 6.9,129.5,128.9,128.8,128.4,128.34,128.26,128.2,127.9,127.6,127.34 ,127.27,126.1,125.5,122.3,121.7,121.6,120.9,119.4,118.8,118.5,111 .0,108.8,97.4,78.9,74.9,73.1,72.1,71.5,70.5,66.8,34.5.HRMS(ESI)m / z calcd for C 48 H 39 O6N3Na[M+Na + ]776.2731found 776.2748.

[0064] Step 4: Preparation of compound 10b:

[0065]

[0066] According to the synthesis method of compound 10a, compound 9b (89.0 mg, 118.0 μmol, 1.0 equiv) was used as a raw material to obtain compound 10b (32.0 mg, 42.4 μmol, 53% for 2 steps) as a yellow-green foam. 1H NMR(400MHz, CDCl3)δ:9.32–9.26(m,1H),9.21(d,J=7.9Hz,1H),7.61–7.40(m,9H),7.40–7.29(m,5H),7 .28–7.23(m,1H),6.98–6.91(m,1H),6.87(t,J=7.4Hz,1H),6.78–6.69(m,2H),6.63(dd,J=5.7,1.5Hz,1 H),5.91–5.82(m,2H),5.34–5.22(m,2H),4.92(d,J=11.8Hz,1H),4.79(s,2H),4.68(d,J=11.8Hz,1H),4 .34(d,J=3.4Hz,1H),3.96–3.88(m,1H),3.49(s,2H),2.91–2.80(m,1H),2.47–2.36(m,1H),2.13(s,3H). 13 C{ 1 H}NMR(101MHz, CDCl3)δ:169.7,169.6,138.6,138.0,137.8,136.8,135.7,131. 1,130.0,129.2,128.9,128.8,128.4,127.8,127.6,127.5,127.0,126.8,126.5, 126.24,126.17,124.1,122.4,120.9,120.6,119.5,118.7,116.8,115.9,111.3, 107.7,91.7,79.3,73.3,71.8,71.4,70.4,69.1,66.7,27.2,27.1.HRMS(ESI)m / z calcd for C 48 H 39 O6N3Na[M+Na + ]776.2731found 776.2744.

[0067] Step 5: Preparation of compound 2:

[0068]

[0069] According to the synthesis method of compound 1, compound 2 (15.0 mg, 32.2 μmol, 76%) was obtained as yellow foam using compound 10b (32.0 mg, 42.4 μmol, 1.0 equiv) as raw material. 1H NMR(400MHz, DMSO-d6)δ:11.03(s,1H),9.19(dd,J=8.1,1.3Hz,1H),9.04(d,J=7.7Hz ,1H),7.87(d,J=8.4Hz,1H),7.77(d,J=8.3Hz,1H),7.63–7.56(m,2H),7.43–7.37(m,2 H),6.91(dd,J=6.0,3.2Hz,1H),6.16(d,J=3.7Hz,1H),4.67(d,J=3.7Hz,1H),4.34(d ,J=3.6Hz,1H),3.94–3.86(m,1H),2.74–2.65(m,1H),2.06(s,3H),2.05–1.98(m,1H). 13 C{ 1 H}NMR(101MHz,DMSO-d6)δ:171.2,171.0,138.9,137.9,130.4,129.3,126.8,126.4,125.0,124.7,123.1,121 .3,120.5,120.3,120.1,119.4,115.3,114.6,112.8,109.5,93.6,79.4,70.1,65.3,30.4,26.4.HRMS(ESI)m / z calcd for C 27 H 20 O7N3[M+HCO2 - ]498.1307found498.1299.

[0070] Example 3: Preparation of Compound 3:

[0071] Step 1: Preparation of compound 7c:

[0072]

[0073] According to the synthesis method of compound 7a, compound 5 (150.0 mg, 274.0 μmol, 1.0 equiv) and compound 6c (811.0 mg, 1.6 mmol, 6.0 equiv) were used as raw materials to obtain compound 7c (120.0 mg, 172.0 μmol, 63% for 3 steps) as a red foam. 1H NMR(400MHz, CDCl3)δ:8.64(d,J=2.9Hz,1H),7.71(d,J=2.8Hz,1H),7.66(s,1H),7.43–7.28(m,11H ),7.25–7.21(m,1H),7.18–7.07(m,3H),6.94–6.85(m,2H),6.80(t,J=8.1Hz,1H),5.57(dd,J=11.3, 2.0Hz,1H),5.22(s,2H),4.75–4.62(m,4H),3.88(dd,J=11.9,2.7Hz,1H),3.82–3.70(m,2H),3.64( s,3H),3.55–3.46(m,1H),3.29(t,J=9.2Hz,1H),2.48–2.38(m,1H),2.16–2.03(m,1H),1.89(s,1H). 13 C{ 1 H}NMR(101MHz, CDCl3)δ:171.73,171.71,137.9,136.1,129.1,128.6,128.5,128.3,128.0,127.8,127.7,126.4,125.0,123.0,122. 8,122.7,122.4,121.3,120.5,111.4,110.2,107.3,107.0,81.2,79.7,79.4,78.0,71.9,71.7,67.3,62.1,61.1,35.6.HRMS(ESI)m / z calcd for C 42 H 39 O7N3Na[M+Na + ]720.2680 found 720.2678.

[0074] Step 2: Preparation of compound 8c:

[0075]

[0076] According to the synthesis method of compound 8a, compound 8c (53.0 mg, 76.2 μmol, 57%) was obtained as yellow-green foam using compound 7c (93.0 mg, 133.3 μmol, 1.0 equiv) as raw material. 1H NMR (400MHz, CDCl3) δ: 11.02 (s, 1H), 9.15 (d, J = 8.0Hz, 1H), 9.00 (d, J = 7.9Hz ,1H),7.55–7.38(m,6H),7.35–7.18(m,10H),5.89(d,J=10.2Hz,1H),5.07(s ,2H),4.72(s,2H),4.49(s,2H),4.37(d,J=10.5Hz,1H),4.25(d,J=11.0Hz,1 H),3.88(t,J=9.0Hz,1H),3.81–3.70(m,5H),3.07(s,1H),2.05–1.89(m,2H). 13 C{ 1 H}NMR(101MHz, CDCl3)δ:169.6,169.0,140.9,139.8,137.8,137.7,129.9,128.5,128.0,127.8,127.6,127.2,127.1,126.1,125.4,122.5,12 2.0,121.3,120.8,120.4,119.2,118.2,118.0,111.5,109.1,82.0,79.1,78.4,77.7,71.7,71.6,66.7,61.4,60.3,35.7.HRMS(ESI)m / zcalcd for C 42 H 37 O7N3Na[M+Na + ]718.2524 found 718.2533.

[0077] Step 5: Preparation of compound 3:

[0078]

[0079] According to the synthesis method of compound 1, compound 8c (25.0 mg, 35.9 μmol, 1.0 equiv) was used as a raw material to obtain compound 3 (14.0 mg, 28.7 μmol, 80%) as a yellow foam. 1H NMR (400MHz, DMSO-d6) δ: 11.81 (s, 1H), 11.13 (s, 1H), 9.20 (d, J = 7.4Hz, 1H), 9.10 (d,J=8.0Hz,1H),8.02(d,J=8.4Hz,1H),7.76(d,J=8.2Hz,1H),7.67–7.56(m,2H) ,7.46–7.35(m,2H),6.64(dd,J=10.0,3.5Hz,1H),6.17(s,1H),5.17(d,J=6.1Hz, 1H),4.08–3.98(m,2H),3.94–3.85(m,2H),3.72–3.63(m,4H),1.90–1.78(m,2H). 13 C{ 1 H}NMR(101MHz,DMSO-d6)δ:171.04,171.00,140.8,134.0,129.4,128.3,127.3,127.1,124.9,124.5,121.6,12 1.5,121.4,121.3,120.5,119.5,118.1,117.0,112.3,80.9,78.7,77.5,70.1,60.3,58.7,38.9.HRMS(ESI)m / z calcd for C 27 H 24 O6N3[M+H + ]486.1660 found 486.1661.

[0080] Example 4: Preparation of Compound 4:

[0081] Step 3: Preparation of compound 9c:

[0082]

[0083] According to the synthesis method of compound 9a, compound 8c (53.0 mg, 76.2 μmol, 1.0 equiv) was used as a raw material to obtain compound 9c (41.0 mg, 60.5 μmol, 78% for 2 steps) as a yellow-green foam. 1HNMR(400MHz, CDCl3)δ:10.01(s,1H),9.36(d,J=7.9Hz,1H),9.26–9.20(m,1H),7.5 7(t,J=7.7Hz,1H),7.52–7.39(m,4H),7.40–7.28(m,9H),7.25–7.19(m,1H),6.98–6. 84(m,2H),5.27(s,2H),4.97(d,J=1.6Hz,1H),4.75(s,2H),4.70(s,2H),4.67(d,J=1 .7Hz,1H),4.23–4.17(m,1H),4.16(d,J=4.0Hz,1H),3.63(s,3H),2.67–2.52(m,2H). 13 C{ 1 H}NMR(101MHz,CDCl3)δ:169.6,169.5,152.5,140.5,140.3,137.8,136.9,1 29.5,128.8,128.40,128.35,128.3,127.9,127.8,127.6,127.33,127.27,1 26.1,125.4,122.2,121.7,121.6,120.9,120.7,119.3,118.7,118.4,110.9 ,108.8,97.2,78.8,77.6,72.9,72.1,71.5,66.8,56.8,34.4.HRMS(ESI)m / z calcd for C 42 H 35 O6N3Na[M+Na + ]700.2418 found 700.2421.

[0084] Step 4: Preparation of compound 10c:

[0085]

[0086] According to the synthesis method of compound 10a, compound 9c (41.0 mg, 60.5 μmol, 1.0 equiv) was used as a raw material to obtain compound 10c (20.7 mg, 30.5 μmol, 51% for 2 steps) as a yellow-green foam. 1H NMR(400MHz, CDCl3)δ:9.34(d,J=7.4Hz,1H),9.22(d,J=7.9Hz,1H),7.68–7.52(m,3H),7.51–7.39 (m,4H),7.40–7.31(m,3H),7.29–7.24(m,1H),6.91–6.82(m,1H),6.79–6.70(m,2H),6.65(dd,J=5 .7,1.7Hz,1H),5.94–5.82(m,2H),5.39–5.28(m,2H),4.81(s,2H),4.14(d,J=3.6Hz,1H),4.00–3. 91(m,1H),3.72(s,3H),3.62–3.48(m,2H),2.84–2.70(m,1H),2.40(d,J=15.3Hz,1H),2.26(s,3H). 13 C{ 1 H}NMR(101MHz, CDCl3)δ:169.7,169.6,138.6,137.9,137.7,135.6,131.2, 123.0,128.3,127.8,127.6,127.4,127.0,126.8,126.5,126.4,126.3,126 .2,124.2,122.4,120.9,120.7,119.5,118.7,116.8,116.0,111.2,107.7, 91.8,79.3,76.1,71.4,70.3,69.1,66.7,59.7,26.83,26.80.HRMS(ESI)m / z Calculate for C 42 H 35 O6N3Na[M+Na + ]700.2418 found 700.2422.

[0087] Step 5: Preparation of compound 4:

[0088]

[0089] According to the synthesis method of compound 1, compound 4 (12.0 mg, 25.9 μmol, 85%) was obtained as yellow foam using compound 10c (20.7 mg, 30.5 μmol, 1.0 equiv) as raw material. 1H NMR (400MHz, DMSO-d6) δ: 11.06 (s, 1H), 9.20 (d, J = 8.0Hz, 1H), 9.05 (d, J = 8.0Hz, 1 H),7.94(d,J=8.5Hz,1H),7.74(d,J=8.3Hz,1H),7.64–7.56(m,2H),7.44–7.38(m ,2H),6.90(dd,J=6.2,2.1Hz,1H),4.62(d,J=3.4Hz,1H),4.11–4.05(m,1H),3.98 (d,J=3.8Hz,1H),3.68(s,3H),2.71–2.62(m,1H),2.12(s,3H),2.10–2.02(m,1H). 13 C{ 1 H}NMR(101MHz,DMSO-d6)δ:171.2,171.00,138.6,137.8,130.5,129.4,126.8,126.6,125.0,124.8,123.1,121.4 ,120.5,120.4,120.2,119.3,115.3,114.6,112.9,109.5,92.0,79.2,78.4,61.7,59.1,30.1,26.7.HRMS(ESI)m / z calcd for C 27 H 22 O5N3[M+H + ]468.1554 found 468.1560.

[0090] Example 5: Anti-leukemia activity test

[0091] (1) Experimental methods:

[0092] Sample preparation: Weigh the sample, add cell-grade DMSO to dissolve and mix, the stock solution concentration is 10mM. Dilute with cell culture medium to 10 times the concentration to be tested.

[0093] Standard operating procedures: (1) Cytotoxicity (CCK-8 method) Detection principle: The detection principle is that the CCK-8 reagent contains a water-soluble tetrazolium salt WST-8, which is reduced to a highly water-soluble yellow formazan product (Formazan) by the dehydrogenase in the cell mitochondria under the action of the electron carrier 1-methoxy-5-methylphenazine dimethyl sulfate (1-Methoxy PMS). The amount of formazan generated is proportional to the number of living cells. (2) Experimental method: ① Inoculation of cells: Use culture medium containing 10% fetal bovine serum to prepare cells into a single cell suspension, and inoculate 90μL 5×10 in each well of a 96-well plate. 4 / mL of adherent cells and 9×104 / mL suspension cells were pre-cultured for 24h under 5% CO2 and 37℃. ② Add the sample solution to be tested: add 10μL sample solution to each well, set 1 concentration for each sample in the initial screening of activity, and set 3 replicate wells; IC 50 8 concentrations (including 0 concentration) were measured, and 3 replicate wells were set for each concentration; the cells were placed in an incubator and cultured for 48 hours. The experiment set up a blank group (Blank), a control group (Control) and a drug group (Drug). ③ Color development: The old culture medium and drug solution were aspirated from the adherent cells (10 μL of CCK-8 solution was directly added to the suspended cells), and 100 μL of CCK-8 solution diluted ten times was added to each well, and cultured for 1 to 4 hours at 37°C and 5% CO2 (operate in dark and observe in real time). ④ Detection: Use an enzyme marker to measure the absorbance at 450nm and record the original data results. ⑤ Use Excel software to standardize the original data, and calculate the cell proliferation inhibition rate through the OD value of each well for the initial screening (formula = (OD Control -OD Drug ) / (OD Control -OD Blank )×100%), and the inhibition rate was calculated. IC 50 The results were calculated by GraphPad Prism8 (version 8.0.2, GraphPad Software Inc), and expressed as ±SD. ⑦ Positive control: Cisplatin.

[0094] (2) Experimental results:

[0095] Table 1. Inhibitory activity of different compounds against four leukemia cell lines

[0096]

[0097] From the data in Table 1, it can be seen that the four indolecarbazole glycoside alkaloids prepared by the present invention have significant anti-leukemia activity. For human myeloid leukemia cells HL-60, the four compounds all have good anti-leukemia activity, IC 50 0.69~1.91μM. For human chronic myeloid leukemia cells K-562, compound 2 (IC 50 =1.68 μM) and 4 (IC 50 =1.90 μM) showed similar antitumor activity to midostaurin (IC 50 =1.78 μM) and compound 1 (IC 50 =1.05 μM) and 3 (IC 50 =0.49 μM) had significantly stronger antitumor activity than midostaurin. All four compounds showed significant antitumor activity against human myelomonocytic leukemia cells MV-4-11 (IC50 =5.6~10.3nM), and the activity was better than midostaurin (IC 50 =11.3 nM). The antitumor activities (IC 50 =0.22~0.85μM) are stronger than midostaurin (IC 50 =0.96 μM).

[0098] Example 6: Study on the mechanism of action of compound 1

[0099] (1) Experimental methods:

[0100] Flow cytometry apoptosis detection:

[0101] 1) Cell culture: Grow cells to an appropriate density and then perform the desired treatment.

[0102] 2) Cell collection: Collect cells by centrifugation and remove the culture medium. Wash the cells to remove residual culture medium or serum.

[0103] 3) Staining: Use fluorescent dyes (such as Annexin V-FITC and PI) in the apoptosis detection kit for staining. Annexin V binds to phosphatidylserine on the cell membrane, while PI is usually used to mark dead cells that have lost membrane integrity.

[0104] 4) Incubation: Incubate at room temperature in the dark, usually for 30 minutes.

[0105] 5) On-machine detection: Set appropriate fluorescence channels on the flow cytometer to detect the fluorescence signals of Annexin V-FITC and PI.

[0106] Flow cytometry cell cycle assay:

[0107] 1) Cell collection: Collect cells by centrifugation and remove the culture medium.

[0108] 2) Fix cells: Fix cells with cold 70% ethanol and store at 4°C overnight. Wash fixed cells with PBS to remove ethanol.

[0109] 3) RNase treatment: Treat cells with RNase to remove RNA and avoid affecting DNA measurement.

[0110] 4) Staining: The cells were stained with the DNA binding dye PI.

[0111] 5) Incubation: Incubate in the dark at room temperature for a certain period of time to allow the dye to fully bind to the DNA.

[0112] 6) On-machine detection: Set appropriate fluorescence channels on the flow cytometer to detect the fluorescence signal of DNA.

[0113] Western Blot Experiment:

[0114] 1) Prepare the upper and lower layers of glue: install glue preparation device and leak detection, and prepare glue.

[0115] 2) Gel running: prepare electrophoresis solution, load samples, and perform electrophoresis.

[0116] 3) Transfer: prepare transfer solution, transfer the membrane, and wash the plate.

[0117] 4) Closed.

[0118] 5) Primary antibody incubation: Cut the membrane and incubate with primary antibody

[0119] 6) Secondary antibody incubation.

[0120] 7) Development.

[0121] (2) Experimental results:

[0122] The experimental groups were as follows: Group A: blank control group: MV-4-11 + compound 1 (0 μM); Group B: MV-4-11 + compound 1 (0.001 μM); Group C: MV-4-11 + compound 1 (0.01 μM); Group D: MV-4-11 + compound 1 (0.1 μM); Group E: positive control group: MV-4-11 + midostaurin (0.01 μM)

[0123] Figure 1 and Figure 2 It shows that 0.001μM compound 1 does not cause apoptosis of MV-4-11 cells, while 0.01μM compound 1 and 0.01μM midostaurin can induce apoptosis of MV-4-11 cells, but not significantly. And the apoptosis effect induced by compound 1 is stronger than that of midostaurin. With the increase of concentration, 0.1μM compound 1 can effectively induce apoptosis of MV-4-11 cells.

[0124] Figure 3It shows that although 0.001μM and 0.01μM compound 1 can lead to a decrease in the proportion of cells in the G0 / G1 phase and S phase and an increase in the proportion of cells in the G2 / M phase, the effect is weak. However, 0.1μM compound 1 and 0.01μM midostaurin can significantly lead to a decrease in the proportion of cells in the G0 / G1 phase and S phase and an increase in the proportion of cells in the G2 / M phase, and the result caused by 0.1μM compound 1 is more obvious than that of 0.01μM midostaurin. Therefore, the experimental results show that compound 1, like midostaurin, exerts its anti-tumor activity by arresting the cell cycle of MV-4-11 cells in the G2 / M phase, causing apoptosis of MV-4-11 cells.

[0125] Figure 4 The results showed that compound 1 significantly inhibited the expression of p-FLT3, p-STAT5, p-AKT and p-Erk in MV4-11 cells in a dose-dependent manner within the range of 6.25 to 100 nM. Therefore, compound 1 may inhibit the growth and proliferation of MV4-11 cells by downregulating the phosphorylation level of the FLT3 pathway and inhibiting the activity of STAT5 downstream transcription factors.

[0126] The preferred embodiments of the present invention are described in detail above, but the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, the technical solution of the present invention can be subjected to a variety of equivalent transformations, and these equivalent transformations all belong to the protection scope of the present invention. It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further explain various possible combinations. In addition, the various different embodiments of the present invention can also be arbitrarily combined, as long as they do not violate the concept of the present invention, they should also be regarded as the contents disclosed by the present invention.

Claims

1. An indolecarbazole glycoside alkaloid or a pharmaceutically acceptable salt thereof as shown in formula 1-4:

2. A method for preparing an indolecarbazole glycoside alkaloid as shown in formula 1-4, characterized in that: The following steps are involved: Among them, R 1 : is H, methyl (CH3) or benzyl (Bn); R 2 : is H, hydroxyl (OH) or benzyloxy (BnO); Step 1: Compounds 5, 6, a phosphine reagent and an azo reagent are dissolved in a first solvent and reacted at -78°C to room temperature for 6 to 24 hours; then, tert-butyloxycarbonyl (Boc) is removed under silica gel / 90°C; and tert-butyldiphenylsilyl (TBDPS) is removed under tetrabutylammonium fluoride (TBAF) conditions to obtain compound 7; Step 2: dissolving compound 7 in a second solvent, adding the first catalyst, and reacting at 60° C. to 110° C. for 6 to 24 hours to obtain compound 8; Step 3: Compound 8, triphenylphosphine, imidazole and iodine are dissolved in a third solvent, and reacted at 60° C. to 110° C. for 1 to 8 hours; then, the olefin is eliminated under 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) conditions to obtain compound 9; Step 4: Compound 9, DBU, potassium iodide and iodine are dissolved in tetrahydrofuran (THF) and reacted at 0°C to room temperature for 10 to 60 minutes; then iodine is removed under the conditions of hydrogen, 20% Pd(OH)2 and N,N-diisopropylethylamine (DIPEA) to obtain compound 10; Step 5: Dissolve compound 8 or 10 in a fourth solvent, add a second catalyst, and react at 0°C to room temperature for 12 to 24 hours under hydrogen conditions; then react at room temperature for 1 hour under ammonia conditions to obtain an indolecarbazole glycoside alkaloid represented by formula 1-4.

3. The method for preparing indolecarbazole glycoside alkaloids according to claim 2, characterized in that: In step 1, the phosphine reagent is selected from any one of compounds P1 to P12; the azo reagent is selected from any one of compounds A1 to A5; the molar ratio of compounds 5, 6, the phosphine reagent and the azo reagent is: 1:3.0:6.0:6.0 to 1:4.0:12.0:12.0; the first solvent is: THF, dichloromethane, toluene or acetonitrile; the concentration of compound 5 in the first solvent is: 0.05 to 0.2 mmol / mL.

4. The method for preparing indolecarbazole glycoside alkaloids as claimed in claim 2, characterized in that: In step 2, the first catalyst is palladium acetate, palladium chloride or copper chloride; the molar ratio of compound 7 to the first catalyst is 1:1.0 to 1:2.0; the second solvent is acetic acid, N,N-dimethylformamide or acetone; the concentration of compound 7 in the second solvent is 0.01 to 0.1 mmol / mL.

5. The method for preparing indolecarbazole glycoside alkaloids as claimed in claim 2, characterized in that: In step 3, the third solvent is THF or PhMe; the concentration of compound 8 in the third solvent is 0.01-0.1 mmol / mL.

6. The method for preparing indolecarbazole glycoside alkaloids according to claim 2, characterized in that: In step 5, the second catalyst is: 10% palladium carbon, 20% palladium hydroxide carbon or 10% palladium carbon / 20% palladium hydroxide carbon; the fourth solvent is THF / methanol, THF / isopropanol or THF / i-PrOH / H2O; the concentration of compound 8 or 10 in the fourth solvent is 0.01-0.1 mmol / mL.

7. Use of the indolecarbazole glycoside alkaloid or a pharmaceutically acceptable salt thereof according to claim 1 in the preparation of an anti-leukemia drug.

8. Use of the indolecarbazole glycoside alkaloid or a pharmaceutically acceptable salt thereof according to claim 1 in the preparation of drugs for anti-myeloid leukemia, anti-myelomonocytic leukemia and anti-lymphocytic leukemia.

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