Indolocarbazole glycoside alkaloids, methods for their preparation and use in anti-leukemia drugs

Abstract

The application belongs to the field of organic synthesis and medicinal chemistry, and particularly relates to a kind of indole carbazole glycoside alkaloids and a preparation method and application in anti-leukemia drugs thereof.The indole carbazole glycoside alkaloids or pharmaceutically acceptable salt thereof, structure as shown in formula 1-4:Pharmacological test proves that the indole carbazole glycoside alkaloids shown in formula 1-4 have anti-leukemia activity, and can be used for preparing anti-myelocytic leukemia, anti-myelomonocytic leukemia, anti-lymphocyte leukemia drugs.

Description

Technical Field

[0001] This invention relates to the fields of organic synthesis and medicinal chemistry, specifically to a class of novel indolecarbazole glycoside alkaloids, pharmaceutical compositions containing them, their preparation methods, and their application in anti-leukemia drugs. Background Technology

[0002] Leukemia is a malignant tumor of the blood system, primarily caused by the arrest of hematopoietic stem cell differentiation, leading to abnormal cell proliferation. In recent years, children have gradually become a high-risk group for leukemia, and its incidence rate is also continuously rising in the adult population. Leukemia generally has a poor prognosis and a high mortality rate, posing a serious threat to people's health and lives. Currently, chemotherapy is the main treatment for leukemia; however, its limited therapeutic effects, severe adverse reactions, and drug resistance remain pressing clinical challenges, making the development of novel drugs for treating leukemia crucial.

[0003] Due to their complex and unique chemical structures and diverse biological activities, indolecarbazole glycoside alkaloids possess significant application value and broad application prospects. Among these compounds, two have entered clinical trials and have 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). Lettaurtinib is an orally bioavailable tyrosine kinase receptor inhibitor with therapeutic effects on AML, chronic myeloid leukemia, and acute lymphoblastic leukemia. Although midostaurin is marketed for the treatment of FLT3-positive AML, its adverse reactions, such as febrile neutropenia, mucositis, and vomiting, severely hinder its clinical application. Lettaurtinib is currently in a phase III clinical trial for acute lymphoblastic leukemia and has not yet been approved for marketing. Therefore, developing novel indolecarbazole glycoside alkaloids that are easy to prepare and have few toxic side effects is of great significance for the treatment of AML. Summary of the Invention

[0004] To address the aforementioned problems, the purpose of this invention is to provide a novel class of indolecarbazole glycoside alkaloids and their preparation method, as well as the application of this class of compounds in anti-leukemia drugs.

[0005] The objective of this invention is achieved through the following technical solution:

[0006] Indolecarbazole glycosides or their pharmaceutically acceptable salts, as shown in Formulas 1-4:

[0007]

[0008] The method for preparing indolecarbazole glycoside alkaloids as shown in Formulas 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: Dissolve compounds 5, 6, phosphine reagent and azo reagent in the first solvent and react at -78°C to room temperature for 6 to 24 hours; then remove the tert-butyloxycarbonyl (Boc) group under silica gel / 90°C conditions; then remove the tert-butyldiphenylsilyl (TBDPS) group under tetrabutylammonium fluoride (TBAF) conditions to obtain compound 7;

[0014] Step 2: Dissolve compound 7 in the second solvent, add the first catalyst, and react at 60°C to 110°C for 6 to 24 hours to obtain compound 8;

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

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

[0017] Step 5: Dissolve compound 8 or 10 in the fourth solvent, add the second catalyst, and react under hydrogen conditions at 0°C to room temperature for 12 to 24 hours; then react under ammonia conditions at room temperature for 1 hour to obtain indolecarbazole glycoside alkaloids as shown in 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 and 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 to 0.1 mmol / mL.

[0023] In step 5, the second catalyst is: 10% palladium on carbon, 20% palladium hydroxide on carbon, or 10% palladium on carbon / 20% palladium hydroxide on carbon, preferably: 10% palladium on 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 indolecarbazole glycoside alkaloids described in this invention are pharmaceutically acceptable salts formed from inorganic or organic acids that are permitted in medicine, 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 tests have demonstrated that the indolecarbazole glycoside alkaloids shown in Formulas 1-4 have anti-leukemia activity and can be used to prepare anti-leukemia drugs.

[0026] The anti-leukemia drugs mentioned are anti-myeloid leukemia, anti-myeloid monocytic leukemia, and anti-lymphocytic leukemia drugs.

[0027] The anti-leukemia activity test used HL-60 anti-human promyelocytic leukemia cells, MV-4-11 anti-human myeloid monocytic leukemia cells, and Jurkat (Clone E6-1) anti-human T-lymphocytic leukemia cells.

[0028] The indolecarbazole glycoside alkaloids provided by this invention exhibit significant anti-leukemia activity. Compounds 1-4 all demonstrate good anti-leukemia activity against human promyelocytic leukemia cells HL-60, with an IC50 value of [missing value]. 50 The concentration ranges from 0.69 to 1.91 μM. Compound 2 (IC50) targets human chronic myeloid leukemia cells K-562. 50 =1.68μM) and 4(IC) 50 =1.90 μM) has antitumor activity similar to midostaurin (IC50). 50 =1.78 μM) equivalent to compound 1 (IC) 50 =1.05μM) and 3(IC 50 Compounds 1-4 exhibited stronger antitumor activity than midostaurin at a concentration of 0.49 μM. Against human myeloid monocytic leukemia cells MV-4-11, compounds 1-4 all showed significant antitumor activity (IC50). 50 =5.6~10.3nM), with activity superior to midostaurin (IC500). 50 =11.3 nM). Antitumor activity (IC50) of compounds 1–4 against human T-lymphoblastic leukemia cells Jurkat (Clone E6-1). 50 =0.22~0.85μM) are all stronger than midostaurin (IC50) 50 =0.96μM).

[0029] The indolecarbazole glycoside alkaloids prepared in this invention exhibit significant anti-leukemia activity with a clearly defined mechanism of action. Further analysis reveals 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 6.25–100 nM. By downregulating the phosphorylation level of the FLT3 pathway and inhibiting the activity of downstream transcription factors of STAT5, it leads to cell arrest in the G2 / M phase of the cell cycle, thereby inducing apoptosis in MV-4-11 cells and exerting anti-leukemia activity. Attached Figure Description

[0030] Figure 1 The figure shows the results of the apoptosis experiment on MV-4-11 cells by compound 1.

[0031] A)-E) are typical flow cytometry results for groups A to E, respectively.

[0032] Figure 2The figure shows the results of the apoptosis experiment on MV-4-11 cells by compound 1.

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

[0034] Figure 3 The figure shows the cell cycle experiment results of compound 1 on MV-4-11 cells.

[0035] Statistical data of MV-4-11 cells in group AE at each cell cycle. Statistical data for each phase are compared with group A. ns represents no significant difference, **p<0.01, ***p<0.001, ****p<0.0001.

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

[0037] The present invention will be further described in detail below through embodiments, but the present invention is not limited to the following embodiments.

[0038] Example 1: Preparation of Compound 1:

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

[0040]

[0041] Triphenylphosphine (2.8 g, 10.8 mmol, 6.0 equiv) and activated... Molecular sieve (2.0 g) was added to a 50.0 mL reaction flask and dissolved in tetrahydrofuran (10.0 mL). A solution of diisopropyl azodicarbonate (DIAD) (2.1 mL, 10.8 mmol, 6.0 equiv) in tetrahydrofuran (5.0 mL) was added at -78 °C, and the mixture was stirred for 1 h. Then, 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) in tetrahydrofuran (5.0 mL) were added at -78 °C, and the mixture was allowed to rise naturally to room temperature for 12 h. After the reaction was complete, the mixture was filtered, diluted with ethyl acetate, washed successively with saturated ammonium chloride solution and saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated by vacuum filtration, and purified by silica gel column chromatography (PE:EA = 8:1) to obtain red foam. Under argon atmosphere, red foam (1.2 g, 984.8 μmol, 1.0 equiv) was added to a 50.0 mL reaction flask, dissolved in toluene (25.0 mL), and then silica gel (2.0 g) was added. The reaction was heated at 90 °C for 12 h. After the reaction was completed, the mixture was filtered, concentrated, and purified by silica gel column chromatography (PE:EA = 4:1) to obtain red foam. Under argon atmosphere, red foam (598.0 mg, 534.7 μmol, 1.0 equiv) was added to a 25.0 mL reaction flask, dissolved in tetrahydrofuran (20.0 mL), and then TBAF (1.6 mL, 1.6 mmol, 3.0 equiv, 1 M in THF) was added at 0 °C. The reaction was carried out at room temperature for 30 min. After the reaction was completed, the solution was diluted with ethyl acetate, washed successively with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated by filtration, and purified by silica gel column chromatography (PE:EA = 2:1) to give compound 7a (400.0 mg, 454.5 μmol, 85%) as a 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] Compound 7a (400.0 mg, 454.5 μmol, 1.0 equiv) was added to a 50.0 mL reaction flask under argon atmosphere, dissolved in 10.0 mL of acetic acid, and then palladium acetate (204.1 mg, 909.0 μmol, 2.0 equiv) was added. The mixture was heated at 70 °C for 8 h. After the reaction was complete, the mixture was filtered, diluted with ethyl acetate, washed successively with saturated sodium bicarbonate solution, water, and saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated by vacuum filtration, and purified by silica gel column chromatography (PE:EA = 4:1) to give compound 8a (308.0 mg, 350.8 μmol, 77%) as a 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] Compound 8a (308.0 mg, 350.8 μmol, 1.0 equiv) was added to a 50.0 mL reaction flask under argon atmosphere, followed by dissolution in tetrahydrofuran (10.0 mL). 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 then added, and the reaction was heated at 80 °C for 2 h. After the reaction was complete, the mixture was concentrated and purified by silica gel column chromatography (PE:EA = 6:1) to obtain a yellow-green foam. Under argon atmosphere, a yellow-green foam (295.0 mg, 298.2 μmol, 1.0 equiv) was added to a 50.0 mL reaction flask, dissolved in tetrahydrofuran (10.0 mL), and then DBU (896.4 μL, 6.0 mmol, 20.0 equiv) was added. The reaction was heated at 80 °C for 1 h. After the reaction was complete, the mixture was diluted with ethyl acetate, washed successively with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated by filtration, and purified by silica gel column chromatography (PE:EA = 8:1) to give compound 9a (172.0 mg, 199.8 μmol, 67%) as a 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 + ]882.3150found 882.3155.

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

[0049]

[0050] Compound 9a (172.0 mg, 199.8 μmol, 1.0 equiv) was added to a 25.0 mL reaction flask under argon atmosphere, dissolved in tetrahydrofuran (5.0 mL), and DBU (60.0 μL, 399.6 μmol, 2.0 equiv) was added at 0 °C. The mixture was stirred for 10 min, then 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 mixture was reacted at 15 °C for 10 min. After the reaction was complete, the mixture was diluted with ethyl acetate, washed successively with saturated sodium thiosulfate solution and saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated by filtration, and purified by silica gel column chromatography (PE:EA = 8:1) to obtain a yellow-green foam. A yellow-green foam (150.0 mg, 151.8 μmol, 1.0 equiv) was added to a 25.0 mL reaction flask, dissolved in tetrahydrofuran (2.0 mL) and methanol (2.0 mL), followed by the addition of N,N-diisopropylethylamine (DIPEA) (79.0 μL, 455.4 μmol, 3.0 equiv) and 20% palladium hydroxide on carbon (10.0 mg). After hydrogen purging, the mixture was stirred at room temperature for 5 h. After the reaction was complete, 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 - ]904.3240found 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 flask, followed by dissolution with tetrahydrofuran (3.0 mL) and isopropanol (3.0 mL). 10% palladium on carbon (50.0 mg) was added, and the mixture was purged with hydrogen. The mixture was stirred at room temperature for 12 h. After the reaction was complete, 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 mixture was stirred at room temperature for 1 h. After the reaction was complete, 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 a 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 + ]470.1347found 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 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 8b (215.3 mg, 0.28 mmol, 58%) was obtained as a yellow-green foam using compound 7b (373.6 mg, 0.48 mmol, 1.0 equiv) as a starting material. 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, using compound 8b (50.0 mg, 64.8 μmol, 1.0 equiv) as a starting material, compound 9b (23.0 mg, 30.5 μmol, 72% for 2 steps) was obtained 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, using compound 9b (89.0 mg, 118.0 μmol, 1.0 equiv) as a starting material, compound 10b (32.0 mg, 42.4 μmol, 53% for 2 steps) was obtained 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 a yellow foam using compound 10b (32.0 mg, 42.4 μmol, 1.0 equiv) as a starting 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 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 a yellow-green foam using compound 7c (93.0 mg, 133.3 μmol, 1.0 equiv) as a starting 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 + ]718.2524found 718.2533.

[0077] Step 5: Preparation of Compound 3:

[0078]

[0079] According to the synthesis method of compound 1, compound 3 (14.0 mg, 28.7 μmol, 80%) was obtained as a yellow foam by using compound 8c (25.0 mg, 35.9 μmol, 1.0 equiv) as a starting material. 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 + ]486.1660found 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, using compound 8c (53.0 mg, 76.2 μmol, 1.0 equiv) as a starting material, compound 9c (41.0 mg, 60.5 μmol, 78% for 2 steps) was obtained 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 + ]700.2418found 700.2421.

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

[0085]

[0086] According to the synthesis method of compound 10a, using compound 9c (41.0 mg, 60.5 μmol, 1.0 equiv) as a starting material, compound 10c (20.7 mg, 30.5 μmol, 51% for 2 steps) was obtained 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 calcd for C 42 H 35 O6N3Na[M+Na + ]700.2418found 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 a yellow foam using compound 10c (20.7 mg, 30.5 μmol, 1.0 equiv) as a starting 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 + ]468.1554found 468.1560.

[0090] Example 5: Anti-leukemia activity test

[0091] (1) Experimental method:

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

[0093] Standard Operating Procedures: (1) Detection Principle of Cytotoxicity (CCK-8 Method): The detection principle is that the CCK-8 reagent contains a water-soluble tetrazolium salt WST-8, which is reduced by dehydrogenases in the cell mitochondria to a highly water-soluble yellow formazan product under the action of the electron carrier 1-methoxy-5-methylphenazine sulfate dimethyl ester (1-Methoxy PMS). The amount of formazan produced is directly proportional to the number of live cells. (2) Experimental Methods: ① Cell Inoculation: Prepare a single-cell suspension with culture medium containing 10% fetal bovine serum, and inoculate 90 μL of 5×10 cells per well of a 96-well plate. 4 adherent cells / mL and 9×104 ① Pre-culture the suspended cells at 5% CO2 and 37℃ for 24 h. ② Add the sample solution: Add 10 μL of sample solution to each well. For the initial activity screening, set one concentration for each sample, with 3 replicates. IC50 50 Eight concentrations (including 0 concentration) were measured, with three replicates for each concentration; cells were incubated for 48 hours. The experiment included a blank group, a control group, and a drug group. ③ Colorimetric development: Adherent cells were aspirated from the old culture medium and drug solution (10 μL of stock CCK-8 solution was added directly to suspension cells), and 100 μL of CCK-8 solution diluted tenfold was added to each well. Cells were incubated at 37℃ and 5% CO2 for 1–4 hours (in the dark, with real-time observation). ④ Detection: The absorbance at 450 nm was measured using a microplate reader, and the raw data results were recorded. ⑤ The raw data were standardized using Excel software. The cell proliferation inhibition rate was initially calculated based on the OD value of each well (formula = (OD...). Control -OD Drug ) / (OD Control -OD Blank (100%), statistical inhibition rate. IC 50 Calculations were performed using GraphPad Prism8 (version 8.0.2, GraphPad Software Inc.), and the results are 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] As shown in Table 1, the four indolecarbazole glycoside alkaloids prepared in this invention exhibit significant anti-leukemia activity. All four compounds showed good anti-leukemia activity against human promyelocytic leukemia cells HL-60, with IC50 values ​​of [missing data]. 50 The concentration ranges from 0.69 to 1.91 μM. Compound 2 (IC50) targets human chronic myeloid leukemia cells K-562. 50 =1.68μM) and 4(IC) 50 =1.90 μM) has antitumor activity similar to midostaurin (IC50). 50 =1.78 μM) equivalent to compound 1 (IC) 50 =1.05μM) and 3(IC 50 The antitumor activity of the four compounds (IC50, 0.49 μM) was significantly stronger than that of midostaurin. All four compounds exhibited significant antitumor activity (IC50, 0.49 μM) against human myeloid monocytic leukemia cells MV-4-11.50 =5.6~10.3nM), with activity superior to midostaurin (IC500). 50 =11.3 nM). Antitumor activity (IC50) of four compounds against human T-lymphoblastic leukemia cells Jurkat (Clone E6-1). 50 =0.22~0.85μM) are all stronger than midostaurin (IC50) 50 =0.96μM).

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

[0099] (1) Experimental method:

[0100] Flow cytometry apoptosis detection:

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

[0102] 2) Cell collection: Cells are collected by centrifugation to remove the culture medium. Cells are washed to remove residual culture medium or serum.

[0103] 3) Staining: Staining was performed using fluorescently labeled dyes (such as Annexin V-FITC and PI) from the apoptosis detection kit. Annexin V binds to phosphatidylserine on the cell membrane, while PI is typically used to label dead cells that have lost membrane integrity.

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

[0105] 5) Detection on flow cytometer: Set up an appropriate fluorescence channel on the flow cytometer to detect the fluorescence signals of Annexin V-FITC and PI.

[0106] Flow cytometry cell cycle detection:

[0107] 1) Cell collection: Cells are collected by centrifugation and the culture medium is removed.

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

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

[0110] 4) Staining: Cells were stained using 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) Detection on flow cytometer: Set up an appropriate fluorescence channel on the flow cytometer to detect the fluorescence signal of DNA.

[0113] Western Blot experiment:

[0114] 1) Prepare the upper and lower layers of adhesive: Install the adhesive dispensing device and perform leak detection, and dispense the adhesive.

[0115] 2) Electrophoresis: 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 + Midotulin (0.01 μM)

[0123] Figure 1 and Figure 2 The results showed that 0.001 μM of compound 1 did not induce apoptosis in MV-4-11 cells, while 0.01 μM of compound 1 and 0.01 μM of midostaurin could induce apoptosis in MV-4-11 cells, but the effect was not significant. Furthermore, the apoptotic effect induced by compound 1 was stronger than that induced by midostaurin. With increasing concentration, 0.1 μM of compound 1 effectively induced apoptosis in MV-4-11 cells.

[0124] Figure 3The results showed that while 0.001 μM and 0.01 μM of compound 1 could reduce the proportion of cells in the G0 / G1 and S phases and increase the proportion of cells in the G2 / M phase, the effects were weak. However, 0.1 μM of compound 1 and 0.01 μM of midostaurin significantly reduced the proportion of cells in the G0 / G1 and S phases and increased the proportion of cells in the G2 / M phase, with the effect of 0.1 μM of compound 1 being more pronounced than that of 0.01 μM of midostaurin. Therefore, the experimental results indicate that compound 1, like midostaurin, exerts its antitumor activity by arresting the cell cycle of MV-4-11 cells in the G2 / M phase, leading to apoptosis.

[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 6.25–100 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 downstream transcription factors of STAT5.

[0126] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details of the above embodiments. Within the scope of the technical concept of the present invention, various equivalent transformations can be made to the technical solutions of the present invention, and these equivalent transformations all fall within the protection scope of the present invention. Furthermore, it should be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. To avoid unnecessary repetition, the present invention will not describe the various possible combinations separately. In addition, various different embodiments of the present invention can also be arbitrarily combined, as long as they do not violate the spirit of the present invention, they should also be considered as the content disclosed by the present invention.

Claims

1. An indolo[3,2-b]carbazole glycoside alkaloid or a pharmaceutically acceptable salt thereof represented by Formula 1-4: Formula 1-4 comprising the following steps: 。 2. The method for preparing indolecarbazole glycoside alkaloids as described in Formulas 1-4 of claim 1, characterized in that, comprising the following steps: ； wherein R 1 : is H, methyl (CH3) or benzyl (Bn); R 2 : is H, hydroxyl (OH) or benzyloxy (BnO); Step 1: dissolving compound 5, 6, a phosphine reagent and an azo reagent in a first solvent, reacting at -78 °C to room temperature for 6-24 hours; then removing tert-butyloxycarbonyl (Boc) under silica gel / 90 °C; and then removing tert-butyldiphenylsilyl (TBDPS) under tetrabutylammonium fluoride (TBAF) to obtain compound 7; Step 2: dissolving compound 7 in a second solvent, adding a first catalyst, and reacting at 60 °C to 110 °C for 6-24 hours to obtain compound 8; Step 3: dissolving compound 8, triphenylphosphine, imidazole and iodine in a third solvent, and reacting at 60 °C to 110 °C for 1-8 hours; then eliminating to olefin under 1, 8-diazabicyclo[5.4.0]undec-7-ene (DBU) 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 10-60 minutes; then the iodine is removed under hydrogen, 20% Pd(OH)2, N , N - DIPEA conditions to give compound 10; Step 5: dissolving compound 8 or 10 in a fourth solvent, adding a second catalyst, and reacting under hydrogen at 0 °C to room temperature for 12-24 hours; and then reacting under ammonia at room temperature for 1 hour to obtain the indolo[3,2-b]carbazole glycoside alkaloid represented by Formula 1-4.

3. The method for preparing indolecarbazole glycoside alkaloids as described in claim 2, characterized in that, In Step 1, the phosphine reagent is selected from any one of compounds P1-P12; the azo reagent is selected from any one of compounds A1-A5; the molar ratio of compound 5, 6, the phosphine reagent and the azo reagent is 1:3.0:6.0:6.0-1:4.0:12.0:12.0; the first solvent is THF, dichloromethane, toluene or acetonitrile; and the concentration of compound 5 in the first solvent is 0.05-0.2 mmol / mL; 。 4. The method for preparing indolecarbazole glycoside alkaloids as described 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 1:1.0-1:2.0; the second solvent is acetic acid, N, N dimethylformamide or acetone; the concentration of compound 7 in the second solvent is 0.01-0.1 mmol / mL.

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

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

7. Use of the indolo[3,2-b]carbazole glycoside alkaloid or a pharmaceutically acceptable salt thereof of claim 1 in the preparation of a drug for resisting myelomonocytic leukemia.

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