CN109879888B - A kind of fred carbazole base A compound and preparation method and application thereof - Google Patents

Abstract

The invention discloses a fred carbazole base A compound and a preparation method and application thereof; the compound is made of staurosporine or halogenated staurosporine through sulfurylation reaction, salification reaction with methyl iodide, and substitution of staurosporine. prepared by reaction and dehydration cyclization reaction; the compound disclosed in the invention has strong selective inhibitory activity on the acute myeloid cell line MV4-11 mutated by Flt3-ITD; weak inhibitory effect on human peripheral blood mononuclear cell PBMC, It can be developed as a drug for the prevention and treatment of leukemia with high efficiency and low toxicity.

Description

A kind of fred carbazole base A compound and preparation method and application thereof

technical field

The invention belongs to the field of medicine, and in particular relates to a fred carbazole base A compound and a preparation method and application thereof.

Background technique

Cancer is the leading cause of death from various diseases in my country, and the morbidity and mortality continue to increase. According to statistics, in 2015, there were 4.292 million new cancer cases and 2.814 million cancer deaths in China, about 12,000 new cancer cases and 7,500 cancer deaths every day; compared with 2010, the cancer incidence in China in 2015 and mortality rates increased by approximately 39% and 44%, respectively, accounting for 22% and 27% of new cancer cases and deaths globally, respectively. Leukemia is one of the ten high-incidence cancers in Chinese men, and the incidence is on the rise (W. Chen, R. Zheng, P. D. Baade, et al. CancerStatistics in China, 2015. CA: Cancer J. Clin. 2016, 66: 115–132).

The onset of acute leukemia is rapid, and it is difficult for patients to undergo bone marrow transplantation in a short period of time. Drug treatment is currently the most effective method. Among them, acute myeloid leukemia (AML) is a blood cancer caused by abnormal proliferation of myeloid leukocytes. Fms-like TyrosineKinase-3 (Flt3) is highly expressed in 17-34% of AML patients (AYH Leung, CH Man, YL Kwong. FLT3 inhibition: a moving and evolving target in acute myeloid leukaemia. Leukemia. 2013, 27: 260–268). Fradcarbazole A is a trace indolocarbazole alkaloid (P Fu, YB Zhuang, Y Wang, PPLiu) obtained from the fermentation product of Streptomyces fradiae007M135. , X. Qi, K. Gu, D. Zhang, WM Zhu. New Indolocarbazoles from a MutantStrain of the Marine-Derived Actinomycete Streptomyces fradiae 007M135, Organic Letter. 2012, 14: 6194–6197); the novelty of its structure and its comparative The good tumor cytotoxic activity has attracted the attention of researchers. The applicant has synthesized Friedcarbazole base A and its three structural analogs, and the three analogs have shown good tumor cytotoxic activity and molecular targets. PKC-β inhibition (LP Wang, XG Mei, CWang, WM Zhu. Biomimetic semi-synthesis of fradcarbazole A and its analogues. Tetrahedron 2015, 71: 7990–7997).

The present invention uses staurosporine as a skeleton to synthesize a new type of Friedcarbazole base A compound. The successful listing of the staurosporine derivative midostaurin as a treatment for AML (approved by the US FDA on April 28, 2017) lays the foundation for the development of such compounds as anti-leukemia drugs. Therefore, compounds synthesized with staurosporine as skeleton may have strong inhibitory activity on acute myeloid cells.

SUMMARY OF THE INVENTION

The purpose of the present invention is to provide a kind of fred carbazole base A compound and its preparation method, as well as the application of the compound in the preparation of medicaments for preventing or treating leukemia.

The purpose of the present invention and the solution to its main technical problems are achieved by adopting the following technical solutions: a Friedcarbazole base A compound, which is prepared on the basis of staurosporine or halogenated staurosporine, and its The structural formula is (I) formula:

(I);

Wherein, R1 is: -H or halogen, R2 is: -H, halogen or methoxy;

The preparation steps of the compound include: sulfurylation reaction, salification reaction with methyl iodide, substitution reaction and dehydration ring closure reaction, specifically:

(1) When R ₁ is selected from -H, the preparation method is as follows: dissolving staurosporine (Ia) in dichloromethane, then adding triethylamine, and performing sulfurylation reaction with N,N'-thiocarbonyldiimidazole; Then, in acetonitrile solvent, salify with iodomethane to obtain compound (Ib); in N,N-dimethylformamide solvent, perform substitution reaction with indoleethylamine to obtain compound (Ic); in dichloromethane solvent, Dehydration and ring closure under the action of trifluoroacetic anhydride and ethanol to obtain Friedcarbazole base A compound (I);

(2) When R ₁ is selected from halogen, the preparation method is as follows: halogenation reaction of staurosporine with halogenated succinimide in methanol or dichloromethane solvent to obtain halogenated staurosporine; and then halogenated staurosporine is obtained. Staurosporine (Ia) is dissolved in dichloromethane, and then triethylamine is added to carry out sulfurylation reaction with N,N'-thiocarbonyldiimidazole; then in acetonitrile solvent, it is salified with iodomethane to obtain compound (Ib) ; Substitution reaction with indoleethylamine in N,N-dimethylformamide solvent to obtain compound (Ic); in dichloromethane solvent, dehydration and ring closure under the action of trifluoroacetic anhydride and ethanol to obtain Furui Decarbazole base A compound, the synthetic technical route is shown in Figure 2.

wherein said R ₁ is selected from -H, Cl, Br, and R ₂ is selected from -H, F, Cl, Br, -OMe.

The application of the Friedcarbazole base A compound in the preparation of a medicament for preventing or treating leukemia. When the compound is used as a medicine, it can be used directly or in the form of a pharmaceutical composition, the pharmaceutical composition contains 0.1-99% of the compound, and the rest is a pharmaceutically acceptable carrier or excipient.

Compared with the prior art, the present invention has obvious advantages and beneficial effects. It can be seen from the above technical solutions that the compounds of the present invention have strong selective inhibitory activity on the acute myeloid cell line MV4-11 mutated by Flt3-ITD; weak inhibitory effect on human peripheral blood mononuclear cell PBMC; Drugs for the prevention and treatment of leukemia.

Description of drawings

Figure 1 is a schematic diagram of the structure of the Fried carbazole base class A compounds.

Figure 2 is a route for synthesizing Fried carbazole base class A compounds.

Figure 3 is the structural formula of compounds 1-14.

Detailed ways

Example 1 Preparation of Compound 1

Under argon protection, staurosporine (1.86 g, 4.0 mmol) was dissolved in 100 mL of dichloromethane, 10 mL of triethylamine and N , N' -thiocarbonyldiimidazole (1.3 g, 8.0 mmol) were added at room temperature, React overnight at room temperature. The reaction solution was poured into 100 mL of ice water, extracted with dichloromethane, the organic phase was dried over anhydrous sodium sulfate and then concentrated, separated by silica gel column chromatography, and eluted with dichloromethane:methanol=20:1 (v/v) to obtain 3'- N -(1-imidazolylthiocarbonyl)staurosporine 1.9 g, 82% yield, ESI-MS m/z 599.2 [M + Na] ⁺ . Under argon protection, 3'- N- (1-imidazolylthiocarbonyl)staurosporine (1.9 g, 3.30 mmol) was dissolved in 200 mL of acetonitrile, and iodomethane (2.0 mL, 33.0 mmol) was added, and the reaction was carried out at room temperature. 24 hours. The reaction solution was directly concentrated, washed with a mixed solution of 200 mL petroleum ether:dichloromethane=4:1 (v/v) to obtain the imidazole of the compound 3'- N- (1-imidazolethiocarbonyl)staurosporine Partial iodomethane salt 1.7 g, 72% yield, ESI-MS m/z 591.2 [M-I] ⁺ . Under argon protection, in a 100 mL two-necked reaction flask, add 400 mg of 5-fluorotryptamine (2.25 mmol), dissolve it in 15 mL of tetrahydrofuran, add 0.5 mL of triethylamine, and add di-tert-butyl dicarbonate (488 at 10°C). mg, 2.25 mmol), reacted at 10 °C for 1 h, added water to terminate the reaction, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, separated by silica gel column chromatography, and eluted with petroleum ether:ethyl acetate (v/v 3:1) to obtain The white powder product was tert-butyl N- [2-(3-(5-fluoroindole))ethyl]carbamate (486 mg, 83% yield). ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.91 (s, 1H, NH), 7.21-7.33 (m, 3H, ArH), 6.87-6.92 (m, 2H, ArH, NH), 3.14-3.19 ( m, 2H, CH ₂ ), 2.76 (t, J = 7.4 Hz, 2H, CH ₂ ), 1.37 (s, 9H, 3×CH ₃ ); ¹³ C NMR (100 MHz, DMSO- d ₆ ) δ 156.6 ( d, ¹ J _CF = 230.6 Hz), 155.6, 132.9, 127.6 (d, ³ J _CF = 9.7 Hz), 124.8, 112.3 (d, ³ J _CF = 9.7 Hz), 112.2(d, ⁴ J _CF = 4.5 Hz) ), 109.0 (d, ² J _CF = 25.2 Hz), 103.0 (d, ² J _CF = 25.2 Hz), 77.5, 40.8, 28.3 (3×C), 25.4; ESI-MS m/z 301.0 [M + Na ] ⁺ . In a 100 mL two-necked reaction flask, add N- [2-(3-(5-fluoroindole))ethyl] tert-butyl carbamate (100 mg, 0.31 mmol) with 11 mL of THF/H ₂ O (10 : 1) was dissolved, DDQ (144 mg, 0.64 mmol) was added at 0 °C, and the reaction was carried out at this temperature for 2 hours. The reaction solution was poured into 100 mL of ethyl acetate, washed with saturated sodium bicarbonate solution until colorless, and ethyl acetate The ester layer was dried over anhydrous sodium sulfate, concentrated, separated by silica gel column chromatography, eluted with petroleum ether:ethyl acetate (v/v 2:1) to obtain N- [2-oxyidene-2-(3-(5 -Fluorindole))ethyl] tert-butyl carbamate (70 mg, 67% yield). ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 8.47 (s, 1H, ArH), 7.81-7.84 (m, 1H, ArH), 7.48-7.51 (m, 1H, ArH), 7.06-7.10 (m, 1H, ArH), 7.03 (t, J = 5.9 Hz, 1H, NH), 4.28 (d, J = 5.9 Hz, 2H, CH ₂ ), 1.40 (s, 9H, 3×CH ₃ ); ¹³ C NMR ( 150 MHz, DMSO- d ₆ ) δ 190.9, 158.6 (d, ¹ J _CF = 235.5 Hz), 156.0, 134.9, 133.0, 126.0 (d, ³ J _CF = 10.8 Hz), 114.1(d, ⁴ J _CF = 4.0 Hz), 113.5 (d, ³ J _CF = 10.8 Hz), 111.1 (d, ² J _CF = 26.0 Hz), 106.0(d, ² J _CF = 26.0 Hz), 77.9, 46.8, 28.3 (3×C); ESI-MS m/z 315.0 [M + Na] ⁺ . Dissolve N- [2-oxyidene-2-(3-(5-fluoroindole))ethyl]carbamic acid tert-butyl ester (300 mg, 0.95 mmol) in 5 mL of trifluoroacetic acid, and react at 10 °C For one hour, benzene (5 mL × 3 times) was added to azeotropically remove trifluoroacetic acid to obtain N- [2-oxyidene-2-(3-(5-fluoroindole))ethyl]ammonium trifluoroacetate 185 mg, 72% yield. The iodomethane salt of the imidazole moiety of 3'- N- (1-imidazolylthio)staurosporine (121 mg, 0.17 mmol) was dissolved in 5 mL of DMF, 0.5 mL of triethylamine and N- [2-oxygen Subunit-2-(3-(5-fluoroindole))ethyl]ammonium trifluoroacetate (147 mg, 0.51 mmol) was reacted at room temperature for 24 hours. The reaction solution was diluted with 10 mL of ethyl acetate, washed with 1N hydrochloric acid, dried over anhydrous sodium sulfate and concentrated. Semi-preparative HPLC separation, MeOH: _H2O =9:1 (v/v) eluting to give 3'- N- [ N- (2-oxoidene-2-(3-(5-fluoroindole)) Ethyl)carbamoyl]staurosporine 61 mg, yield 51%. ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 12.15 (s, 1H, NH), 9.29 (d, J = 7.9 Hz, 1H, ArH), 8.61 (s, 1H, NH), 8.58 (s, 1H , ArH), 8.06(d, J = 7.7 Hz, 1H, ArH), 8.01 (d, J = 8.5 Hz, 1H, ArH), 7.94 (brs, 1H, NH), 7.85 (dd, J = 9.8, 2.4 Hz, 1H, ArH), 7.75 (d, J = 8.2 Hz, 1H, ArH), 7.48-7.54(m, 3H, ArH), 7.37 (t, J = 7.4 Hz, 1H, ArH), 7.31 (t, J = 7.4 Hz, 1H, ArH), 7.08-7.12 (m, 2H, ArH, H-1'), 5.92 (brs, 1H, H-3'), 4.95-5.05 (m, 4H, H-7, H-3''), 4.53 (brs, 1H, H-4'), 2.95 (s, 3H, 3'-NCH ₃ ), 2.87 (s, 3H, 4'-OCH ₃ ), 2.71-2.76 (m , 1H, H-2'a), 2.36 (s, 3H, 6'-CH ₃ ), 2.27-2.33 (m, 1H, H-2'b); ¹³ CNMR (150 MHz, DMSO- d ₆ ) δ 190.2, 182.5, 172.0, 158.6 (d, ¹ J _CF = 233.5 Hz), 139.1, 136.4, 134.8, 133.0, 132.8, 129.1, 126.0 (d, ³ J _CF = 10.0 Hz), 125.7, 125.4, 125 123.8, 122.7, 121.5, 120.4, 119.5, 119.4, 115.3, 114.3(d, ⁴ J _CF = 4.2 Hz), 114.2, 113.9, 113.5 (d, ³ J _CF = 10.0 Hz), 111.1 (d, ² J _CF = 4.2 Hz) 24.0 Hz), 109.2, 106.0 (d, ² J _CF = 24.0 Hz), 95.0, 82.9, 82.4, 60.4, 54.3, 51.9, 45.5, 32.8, 29.6, 27.7 ; ESI-MS m/z 723.2 [M + Na] ⁺ . Under the protection of argon, in a 25 mL two-necked reaction flask, add 3'- N- [ N- (2-oxyidene-2-(3-(5-fluoroindole))ethyl)aminothiocarbonyl ] Staurosporine (55 mg, 0.079 mmol) was dissolved in 6 mL of dichloromethane and 200 µL of ethanol, and 400 µL of trifluoroacetic anhydride was added at 0°C. After 1 hour of reaction, water was added to terminate the reaction, and extracted with dichloromethane. After drying with sodium sulfate, concentrated, semi-preparative HPLC separation, MeOH:H ₂ O=9:1 (v/v) elution to obtain 34 mg of 5'''-fluorofred carbazole base (compound 1), which was collected rate 63%. ¹ HNMR (600 MHz, DMSO- d ₆ ) δ 11.46 (s, 1H, NH), 9.32 (d, J = 7.8 Hz, 1H, ArH), 8.62 (s, 1H, NH), 8.06 (d, J = 8.0 Hz, 1H, ArH), 8.02 (d, J = 8.0 Hz, 1H, ArH), 7.66-7.68 (m, 2H, ArH), 7.54-7.58 (m, 2H, ArH), 7.44-7.51 (m, 3H, ArH), 7.35 (t, J = 7.2 Hz, 1H, ArH), 7.31 (t, J = 7.3 Hz, 1H, ArH), 7.08 (t, J =7.2 Hz, 1H, ArH), 7.04 (t , J = 8.4 Hz, 1H, H-1'), 4.96-5.01 (m, 3H, H-7, H-3'), 4.49 (s, 1H, H-4'), 2.89 (s, 3H, 3'-NCH ₃ ), 2.85-2.88 (m, 1H, H-2'a), 2.71 (s, 3H, 4'-OCH ₃ ), 2.43 (s, 3H, 6'-CH ₃ ), 2.38- 2.42 (m, 1H, H-2'b); ¹³ C NMR (150 MHz, DMSO- d ₆ ) δ 171.9, 167.5, 157.4 (d, ¹ J _CF = 232.7 Hz), 138.8, 136.3, 134.1, 133.2, 132.7, 129.3, 125.7, 125.4, 125.1, 125.0, 124.9 (d, ³ j _CF = 9.5Hz), 124.9, 123.8, 122.7, 120.4, 119.5, 119.4, 115.2, 113.6, 113.1 (D, D, ³ J _CF = 9.8 Hz), 110.1(d, ² J _CF = 24.1 Hz), 109.0, 107.2 (d, ⁴ J _CF = 4.9 Hz), 104.0 (d, ² J _CF = 24.1 Hz), 94.8, 82.5, 82.4, 60.2, 53.2, 45.5, 34.5, 29.3, 27.1; HRESI-MS m/z 683.2236 [M + H] ⁺ (calcd for C ₃₉ H _32N6O3FS , _{683.2235 )} .

Example 2 Preparation of Compound 2

Under argon protection, in a 100 mL two-necked reaction flask, add 5-chlorotryptamine (300 mg, 1.55 mmol), dissolve it in 15 mL of tetrahydrofuran, add 0.5 mL of triethylamine, and add di-tert-butyl dicarbonate (349 mL) at 10 °C. mg, 1.60 mmol), reacted at 10 °C for 1 h, added water to terminate the reaction, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, separated by silica gel column chromatography, and eluted with petroleum ether: ethyl acetate (v/v 3:1) to obtain White powder product tert-butyl N- [2-(3-(5-chloroindole))ethyl]carbamate (411 mg, 90% yield). ¹ H NMR (600 MHz, CDCl ₃ ) δ 8.40 (s, 1H, NH), 7.53 (d, J = 1.5 Hz, 1H, ArH), 7.24 (d, J = 8.6 Hz, 1H, ArH), 7.12 ( dd, J = 8.6, 1.5 Hz, 1H, ArH), 7.00 (s, 1H, ArH), 4.65 (brs, 1H, NH), 3.35-3.44 (m, 2H, CH ₂ ), 2.88 (t, J = 6.4 Hz, 2H, CH ₂ ), 1.44 (s, 9H, 3×CH ₃ ); ¹³ C NMR (150 MHz, CDCl ₃ ) δ 156.0, 134.7, 128.5, 125.1, 123.4, 122.3, 118.3, 112.9, 112.2, 79.3, 41.0, 28.4 (3×C), 25.6; ESI-MS m/z 317.0 [M + Na] ⁺ . In a 100 mL two-neck reaction flask, add N- [2-(3-(5-chloroindole))ethyl]carbamate tert-butyl ester (380 mg, 1.29 mmol) with 11 mL THF/H ₂ O (10 : 1) was dissolved, DDQ (586 mg, 2.58 mmol) was added at 0 °C, and the reaction was carried out at this temperature for 2 hours. The reaction solution was poured into 100 mL of ethyl acetate, washed with saturated sodium bicarbonate solution until colorless, and ethyl acetate The ester layer was dried over anhydrous sodium sulfate, concentrated, and separated by silica gel column chromatography, eluted with petroleum ether:ethyl acetate (v/v 1:1) to obtain N- [2-oxyidene-2-(3-(5). - chloroindole))ethyl] tert-butyl carbamate (268 mg, 67% yield). ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 12.16 (s, 1H, NH), 8.47 (s, 1H, ArH), 8.14 (s, 1H, ArH), 7.51 (d, J = 8.6 Hz, 1H ), 7.24 (d, J = 8.6 Hz, 1H), 7.03 (t, J = 5.9 Hz, 1H, NH), 4.28 (d, J = 5.9 Hz, 2H, CH ₂ ), 1.40 (s, 9H, 3 ×CH ₃ ); ¹³ C NMR (150 MHz, DMSO- d ₆ ) δ 190.9, 156.0, 134.9, 134.7, 126.5, 126.5, 122.9, 120.2, 113.8, 113.5, 77.9, 46.8, 28.2(3×C); ESI -MS m/z 331.0 [M + Na] ⁺ . N- [2-Oxyidene-2-(3-(5-chloroindole))ethyl] tert-butyl carbamate (230 mg, 0.75 mmol) was dissolved in 5 mL of trifluoroacetic acid and reacted at 10 °C For one hour, benzene (5 mL × 3 times) was added to azeotropically remove trifluoroacetic acid to obtain N- [2-oxyidene-2-(3-(5-chloroindole))ethyl]ammonium trifluoroacetate 202 mg, 88% yield. The iodomethane salt of the imidazole moiety of 3'- N- (1-imidazolylthio)staurosporine (200 mg, 0.28 mmol) was dissolved in 5 mL of DMF, triethylamine 0.5 mL and N- [2-oxygen Subunit-2-(3-(5-chloroindole))ethyl]ammonium trifluoroacetate (85 mg, 0.28 mmol) was reacted at room temperature for 24 hours. The reaction solution was diluted with 10 mL of ethyl acetate, washed with 1N hydrochloric acid, dried over anhydrous sodium sulfate and concentrated. Semi-preparative HPLC separation, MeOH: _H2O =9:1 (v/v) eluting to give 3'- N- [ N- (2-oxoidene-2-(3-(5-chloroindole)) Ethyl)carbamoyl]staurosporine 81 mg, yield 40%. ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 12.22 (s, 1H, NH), 9.30 (d, J = 7.9 Hz, 1H, ArH), 8.61 (s, 1H, NH), 8.59 (s, 1H , ArH), 8.17 (d, J = 2.0 Hz, 1H, ArH), 8.05 (t, J = 10.3 Hz, 1H, ArH), 7.99 (t, J =9.6 Hz, 1H, ArH), 7.95 (brs, 1H, NH), 7.74 (d, J = 8.3 Hz, 1H, ArH), 7.54 (d, J = 8.6 Hz, 1H, ArH), 7.47-7.50 (m, 2H, ArH), 7.36 (t, J = 7.5 Hz, 1H, ArH), 7.31 (t, J = 7.5 Hz, 1H, ArH), 7.26 (dd, J = 8.6, 2.0 Hz, 1H, ArH), 7.09 (t, J = 7.7 Hz, 1H, H -1'), 5.83 (brs, 1H, H-3'), 4.94-5.07 (m, 4H, H-7, H-3''), 4.53 (brs, 1H, H-4'), 2.95 ( s, 3H, 3'-NCH ₃ ), 2.87 (s, 3H, 4'-OCH ₃ ), 2.71-2.75(m, 1H, H-2'a), 2.36 (s, 3H, 6'-CH ₃ ), 2.27-2.33 (m, 1H, H-2'b); ¹³ C NMR (150MHz, DMSO- d ₆ ) δ 190.2, 182.5, 171.9, 139.1, 136.3, 134.9, 134.6, 132.7, 129.1, 126.6, 126.5 , 125.7, 125.4, 125.0, 124.9, 123.8, 122.9, 122.7, 121.4,120.3, 120.3, 119.5, 115.3, 115.2, 113.9, 113.8, 95.0, 82.3, 54.3, 51.9, 45.5.5.5.5.5.5.5.5.5.5.5.5.5.5.9 , 32.9, 29.5, 27.7; ESI-MS m/z 739.2 [M +Na] ⁺ . Under the protection of argon, in a 25 mL two-necked reaction flask, add 3'- N- [ N- (2-oxyidene-2-(3-(5-chloroindole))ethyl)aminothiocarbonyl ] Staurosporine (60 mg, 0.084 mmol) was dissolved in 6 mL of dichloromethane and 200 µL of ethanol, and 400 µL of trifluoroacetic anhydride was added at 0°C. After 1 hour of reaction, water was added to terminate the reaction, and extracted with dichloromethane. It was dried with sodium sulfate, concentrated, separated by semi-preparative HPLC, and eluted with MeOH:H ₂ O=9:1 (v/v) to obtain 29 mg of 5'''-chlorofred carbazole base (compound 2), which was collected rate 49%. ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 11.57 (s, 1H, NH), 9.32 (d, J = 7.8 Hz, 1H, ArH), 8.63 (s, 1H, NH), 8.07 (d, J = 7.8 Hz, 1H, ArH), 8.03 (d, J = 8.5 Hz, 1H, ArH), 7.80 (s, 1H, ArH), 7.68 (brs, 1H, ArH), 7.58 (brs, 1H, ArH), 7.47-7.51 (m, 3H, ArH), 7.36 (t, J = 7.4 Hz, 1H, ArH), 7.32 (t, J = 7.5Hz, 1H, ArH), 7.18-7.20 (m, 1H, ArH), 7.09 (t, J = 7.4 Hz, 1H, H-1'), 4.97-5.01 (m, 3H, H-7, H-3'), 4.50 (s, 1H, H-4'), 2.91 (s , 3H, 3'-NCH ₃ ), 2.85-2.90(m, 1H, H-2'a), 2.71 (s, 3H, 4'-OCH ₃ ), 2.43 (s, 3H, 6'-CH ₃ ) , 2.39-2.43 (m, 1H,H-2'b); ¹³ C NMR (150 MHz, DMSO- d ₆ ) δ 171.9, 167.7, 138.7, 136.3, 135.0, 134.5, 132.7, 129.2, 125.9, 125.7, 125.4 , 125.1, 125.0, 124.6, 124.3, 123.8, 122.7, 121.9, 121.5, 120.3, 119.5 (2×C), 119.4, 118.2, 115.2, 114.1, 113.6 (2×C), 109.4 (2×C), 8.4, 8.206 , 60.2, 53.2, 45.4, 34.5, 29.2, 27.1; HRESI-MS m/z _699.1940 [ _M +H] ⁺ (calcd for _{C39H32N6O3ClS} , _699.1940 ).

Example 3: Preparation of Compound 3

Under the protection of argon, in a 100 mL two-necked reaction flask, add 5-bromotryptamine (300 mg, 1.26 mmol), dissolve it in 15 mL of tetrahydrofuran, add 0.5 mL of triethylamine, and add di-tert-butyl dicarbonate (285 mL at 10°C). mg, 1.31 mmol), reacted at 10 °C for 1 h, added water to terminate the reaction, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, separated by silica gel column chromatography, and eluted with petroleum ether: ethyl acetate (v/v 3:1) to obtain White powder product tert-butyl N- [2-(3-(5-bromoindole))ethyl]carbamate (359 mg, 84% yield). ¹ H NMR (600 MHz, CDCl ₃ ) δ 8.44 (s, 1H, NH), 7.73 (d, J = 1.5 Hz, 1H, ArH), 7.27-7.30 (m, 1H, ArH), 7.25 (d, J = 8.6 Hz, 1H, ArH), 7.02 (s, 1H, ArH), 4.69 (brs, 1H, NH), 3.37-3.47 (m, 2H, CH ₂ ), 2.92 (t, J = 6.6 Hz, 2H, CH ₂ ), 1.48 (s, 9H, 3×CH ₃ ); ¹³ C NMR (150 MHz, CDCl ₃ ) δ 156.0, 135.0, 129.2, 124.8, 123.3, 121.4, 112.9, 112.6 (2×C), 79.3, 41.0, 28.4 (3×C), 25.6; ESI-MS m/z 361.0 [M + Na] ⁺ . In a 100 mL two-neck reaction flask, add N- [2-(3-(5-bromoindole))ethyl] tert-butyl carbamate (100 mg, 0.296 mmol) with 11 mL THF/H ₂ O (10 : 1) was dissolved, DDQ (134 mg, 0.592 mmol) was added at 0 °C, and the reaction was carried out at this temperature for 2 hours. The reaction solution was poured into 100 mL of ethyl acetate, washed with saturated sodium bicarbonate solution until colorless, and ethyl acetate The ester layer was dried over anhydrous sodium sulfate, concentrated, and separated by silica gel column chromatography, eluted with petroleum ether:ethyl acetate (v/v 1:1) to obtain N- [2-oxyidene-2-(3-(5). -Bromoindole))ethyl] tert-butyl carbamate (61 mg, 58% yield). ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 8.45 (s, 1H, ArH), 8.30 (d, J = 1.5 Hz, 1H, ArH), 7.46 (d, J = 8.6 Hz, 1H), 7.35 ( dd, J = 8.6, 1.5 Hz, 1H), 7.03 (t, J =5.9 Hz, 1H, NH), 4.29 (d, J = 5.9 Hz, 2H, CH ₂ ), 1.40 (s, 9H, 3×CH ESI _- MS ^_ _{_} m/z 375.0 [M + Na] ⁺ . N- [2-Oxyidene-2-(3-(5-bromoindole))ethyl] tert-butyl carbamate (61 mg, 0.17 mmol) was dissolved in 5 mL of trifluoroacetic acid and reacted at 10 °C For one hour, benzene (5 mL × 3 times) was added to azeotropically remove trifluoroacetic acid to obtain N- [2-oxyidene-2-(3-(5-bromoindole))ethyl]ammonium trifluoroacetate 56 mg, 94% yield. The iodomethane salt of the imidazole moiety of 3'- N- (1-imidazolylthio)staurosporine (185 mg, 0.26 mmol) was dissolved in 5 mL of DMF, triethylamine 0.5 mL and N- [2-oxygen Subunit-2-(3-(5-bromoindole))ethyl]ammonium trifluoroacetate (180 mg, 0.52 mmol,) was reacted at room temperature for 24 hours. The reaction solution was diluted with 10 mL of ethyl acetate, washed with 1N hydrochloric acid, dried over anhydrous sodium sulfate and concentrated. Semi-preparative HPLC separation, MeOH: _H2O =9:1 (v/v) eluting to give 3'- N- [ N- (2-oxyidene-2-(3-(5-bromoindole)) Ethyl)carbamoyl]staurosporine 91 mg, yield 46%. ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 12.22 (s, 1H, NH), 9.29 (d, J = 7.9 Hz, 1H, ArH), 8.61 (s, 1H, NH), 8.56 (s, 1H , ArH), 8.31 (s, 1H, ArH), 8.05 (d, J = 7.7 Hz, 1H, ArH), 8.00 (d, J = 8.5 Hz, 1H, ArH), 7.95(brs, 1H, NH), 7.74 (d, J = 8.3 Hz, 1H, ArH), 7.48-7.50 (m, 3H, ArH), 7.35-7.38 (m, 2H, ArH), 7.30 (t, J = 7.5 Hz, 1H, ArH), 7.08 (t, J = 7.7 Hz, 1H, H-1'), 5.90 (brs, 1H, H-3'), 4.92-5.06 (m, 4H, H-7, H-3''), 4.52 ( brs, 1H, H-4'), 2.94 (s, 3H, 3'-NCH ₃ ), 2.87 (s, 3H, 4'-OCH ₃ ), 2.70-2.74 (m, 1H, H-2'a) , 2.35 (s, 3H, 6'-CH ₃ ), 2.26-2.32 (m, 1H, H-2'b); ¹³ C NMR (150 MHz, DMSO- d ₆ ) δ 190.3, 182.5, 172.0, 139.1, 136.4, 135.2, 134.5, 132.8, 129.1, 127.3, 125.7,125.5, 125.4, 125.1, 124.9, 123.8, 123.3, 122.7, 121.5, 120.4, 119.5, 119.4,115.3, 114.6, 114.4, 114.2, 113.9, 113.7, 109.2, 95.0, 82.9, 82.4, 60.5, 54.3, 51.9, 45.5, 32.9, 29.7, 27.7; ESI-MS m/z 783.2[M+Na] ⁺ . Under the protection of argon, in a 25 mL two-necked reaction flask, add 3'- N- [ N- (2-oxyidene-2-(3-(5-bromoindole))ethyl)aminothiocarbonyl ] Staurosporine (42 mg, 0.055 mmol) was dissolved in 6 mL of dichloromethane and 200 µL of ethanol, and 400 µL of trifluoroacetic anhydride was added at 0°C. After 1 hour of reaction, water was added to terminate the reaction, and extracted with dichloromethane. It was dried over sodium sulfate, concentrated, separated by semi-preparative HPLC, eluted with MeOH:H ₂ O=9:1 (v/v) to obtain 28 mg of 5'''-bromofried carbazole base (compound 3), which was collected rate of 69%. ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 11.58 (s, 1H, NH), 9.32 (d, J = 3.0 Hz, 1H, ArH), 8.62 (s, 1H, NH), 8.06 (d, J = 7.4 Hz, 1H, ArH), 8.02 (d, J = 7.9 Hz, 1H, ArH), 7.93 (s, 1H, ArH), 7.65-7.68 (m, 2H, ArH), 7.56 (s, 1H, ArH) ), 7.46-7.51 (m, 2H, ArH), 7.43 (d, J = 8.5 Hz, 1H, ArH), 7.35 (t, J = 6.3 Hz, 1H, ArH), 7.29-7.32 (m, 2H, ArH) ), 7.09 (brs, 1H, H-1'), 4.98-5.01 (m, 3H, H-7, H-3'), 4.49 (s, 1H, H-4'), 2.91 (s, 3H, 3'-NCH ₃ ), 2.85-2.90 (m, 1H, H-2'a), 2.71 (s, 3H, 4'-OCH ₃ ), 2.43 (s, 3H, 6'-CH ₃ ), 2.38- 2.43 (m, 1H, H-2'b); ¹³ C NMR (150 MHz, DMSO- d ₆ ) δ 171.9, 167.8, 138.7, 136.3, 135.2, 134.5, 132.7, 129.2, 126.6, 125.7, 125.4, 125.1, 125.0, 124.5, 124.4, 123.8, 122.7, 121.5, 121.2,120.3, 119.5, 119.4 (2×C), 115.2, 114.2, 114.0, 113.5, 112.3, 109.0, 106.6,94.8, 82.5, 82.4, 60.2, 53.2, 45.4 , 34.5, 29.2, 27.1; HRESI-MS m/z _743.1432 [ _M +H] ⁺ (calcd for _{C39H32N6O3BrS} , _743.1434 ).

Example 4 Preparation of Compound 4

Under argon protection, in a 100 mL two-necked reaction flask, add 5-methoxytryptamine (1.0 g, 5.26 mmol), dissolve it in 15 mL of tetrahydrofuran, add 1.5 mL of triethylamine, and add di-tert-butyl dicarbonate at 10 °C (1.37 g, 6.32 mmol), reacted at 10 °C for 1 h, added water to terminate the reaction, extracted with ethyl acetate, dried over anhydrous sodium sulfate and concentrated, separated by silica gel column chromatography, washed with petroleum ether: ethyl acetate (v/v 5:1) A white powder product was obtained from tert-butyl N- [2-(3-(5-methoxyindole))ethyl]carbamate (1.51 g, yield 99%). ¹ H NMR (600 MHz, CDCl ₃ ) δ 8.18 (s, 1H, NH), 7.27 (d, J = 8.8 Hz, 1H, ArH), 7.06 (s, 1H, ArH), 7.01 (s, 1H, ArH) ), 6.89(dd, J = 8.8, 1.5 Hz, 1H, ArH), 4.61 (brs, 1H, NH), 3.89 (s, 3H, OMe), 3.46-3.50 (m, 2H, CH ₂ ), 2.94 ( t, J = 6.4 Hz, 2H, CH ₂ ), 1.46 (s, 9H, 3×CH ₃ ); ¹³ C NMR (150 MHz, CDCl ₃ ) δ 156.0, 154.0, 131.5, 127.7, 122.8, 112.8, 112.3, 111.9, 100.6, 79.1, 55.9, 40.7, 28.4 (3×C), 25.8; ESI-MS m/z 313.1 [M + Na] ⁺ . In a 100 mL two-necked reaction flask, N- [2-(3-(5-methoxyindole))ethyl] tert-butyl carbamate (1.3 g, 4.48 mmol) was added with 55 mL of THF/H ₂ O ( 10:1) was dissolved, DDQ (2.0 g, 8.96 mmol) was added at 0 °C, the reaction was carried out at this temperature for 2 hours, the reaction solution was poured into 100 mL of ethyl acetate, washed with saturated sodium bicarbonate solution until colorless, acetic acid The ethyl ester layer was dried with anhydrous sodium sulfate and then concentrated, and separated by silica gel column chromatography, eluted with petroleum ether:ethyl acetate (v/v 1:1) to obtain N- [2-oxyidene-2-(3-( 5-Methoxyindole))ethyl]carbamate tert-butyl ester (1.1 g, 81% yield). ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 11.86 (brs, 1H, NH), 8.33 (d, J =3.1 Hz, 1H, ArH), 7.67 (d, J =2.5 Hz, 1H, ArH), 7.37 (d, J = 8.8 Hz, 1H), 6.97 (t, J = 5.9 Hz, 1H, NH), 6.85 (dd, J = 8.8, 2.5 Hz, 1H), 4.26 (d, J = 5.9 Hz, 2H) , CH ₂ ), 3.78 (s, 3H, OMe), 1.41 (s, 9H, 3×CH ₃ ); ¹³ C NMR (150 MHz, DMSO- d ₆ ) δ 190.7, 156.0, 155.4, 133.4, 131.2, 126.2 , 113.8, 112.9, 112.7, 102.9, 77.8, 55.2, 46.7, 28.3(3×C); ESI-MS m/z 327.1 [M + Na] ⁺ . Dissolve tert-butyl N- [2-oxyidene-2-(3-(5-methoxyindole))ethyl]carbamate (1.1 g, 3.62 mmol) in 10 mL of trifluoroacetic acid, 10 °C The reaction was continued for one hour, and benzene (5 mL × 3 times) was added to azeotropically remove the trifluoroacetic acid to obtain N- [2-oxyidene-2-(3-(5-methoxyindole))ethyl]trifluoroacetic acid. Ammonium fluoroacetate 0.9 g, yield 83%. The iodomethane salt of the imidazole moiety of 3'- N- (1-imidazolethiocarbonyl)staurosporine (200 mg, 0.28 mmol) was dissolved in 5 mL of DMF, and 0.5 mL of triethylamine and N- [2-oxygen Subunit-2-(3-(5-methoxyindole))ethyl]ammonium trifluoroacetate (169 mg, 0.56 mmol,) was reacted at room temperature for 24 hours. The reaction solution was diluted with 10 mL of ethyl acetate, washed with 1N hydrochloric acid, dried over anhydrous sodium sulfate and concentrated. Semi-preparative HPLC separation, MeOH:H ₂ O=9:1 (v/v) elution to give 3'- N- [ N- (2-oxyidene-2-(3-(5-methoxyindole) ))ethyl)carbamoyl]staurosporine 96 mg, yield 48%. ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 11.93 (d, J = 3.0 Hz, 1H, NH), 9.30 (d, J =8.0 Hz, 1H, ArH), 8.63 (s, 1H, NH), 8.45 (d, J = 3.0 Hz, 1H, ArH), 8.06 (d, J = 7.8 Hz, 1H, ArH), 8.01 (d, J = 8.5 Hz, 1H, ArH), 7.90 (brs, 1H, NH) , 7.74(d, J = 8.3 Hz, 1H, ArH), 7.70 (d, J = 2.5 Hz, 1H, ArH), 7.46-7.51 (m, 2H, ArH), 7.41 (d, J = 8.8 Hz, 1H , ArH), 7.36 (t, J = 7.4 Hz, 1H, ArH), 7.31 (t, J = 7.5 Hz, 1H, ArH), 7.10 (t, J = 7.6 Hz, 1H, H-1'), 6.87 (dd, J = 8.8, 2.5Hz, 1H, ArH), 5.94 (brs, 1H, H-3'), 4.94-5.05 (m, 4H, H-7, H-3''), 4.52 (brs, 1H, H-4'), 3.79 (s, 3H, 5'''-OMe), 2.96 (s, 3H, 3'-NCH ₃ ), 2.85 (s, 3H, 4'-OCH ₃ ), 2.71- 2.76 (m, 1H, H-2'a), 2.37 (s, 3H, 6'-CH ₃ ), 2.27-2.30 (m, 1H, H-2'b); ¹³ C NMR (150 MHz, DMSO- D ₆ ) Δ 190.0, 182.4, 172.0, 155.5, 139.0, 136.4,133.5, 132.8, 131.3, 129.2, 126.3, 125.4, 125.0, 123.8, 122.7, 120.4, 119.4, 115.3, 114.2, 114.2, 114.2, 115.3, 114.2, 114.2, 114.2, 114.2, 115.3, 114.2, 114.0, 113.9, 113.0, 112.7, 109.2, 102.9, 95.0, 83.0, 82.4, 60.4, 55.3, 54.3, 51.9, 45.5, 32.8, 29.5, 27.7; ESI-MS m /z 735.2[M + Na] ⁺ . Under the protection of argon, in a 25 mL two-necked reaction flask, add 3'- N- [ N- (2-oxyidene-2-(3-(5-methoxyindole))ethyl)aminothio Formyl]staurosporine (38 mg, 0.053 mmol) was dissolved in 6 mL of dichloromethane and 200 µL of ethanol, and 400 µL of trifluoroacetic anhydride was added at 0°C. After 1 hour of reaction, water was added to terminate the reaction. Dichloromethane Extraction, drying over anhydrous sodium sulfate, concentration, semi-preparative HPLC separation, MeOH:H ₂ O=9:1 (v/v) elution to obtain 5''-methoxyfredcarbazole base A (compound 4) 30 mg, yield 79%. ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 11.23 (s, 1H, NH), 9.31 (d, J = 8.0 Hz, 1H, ArH), 8.65 (s, 1H, NH), 8.06 (d, J =7.7 Hz, 1H, ArH), 8.02 (d, J = 8.5 Hz, 1H, ArH), 7.67 (d, J = 8.2 Hz, 1H, ArH), 7.57 (s, 1H, ArH), 7.53 (d, J = 2.0 Hz, 1H, ArH), 7.45-7.51 (m, 2H, ArH), 7.30-7.36 (m, 3H, ArH), 7.26 (brs, 1H, ArH), 7.07-7.10 (m, 1H, H -1'), 6.84 (dd, J = 8.8, 2.0 Hz, 1H, ArH), 5.02 (s, 2H, H-7), 4.97 (d, J = 12.2 Hz, 1H, H-3'), 4.50 (brs, 1H, H-4'), 3.84 (s, 3H, 5'''-OMe), 2.91 (s, 3H, 3'-NCH ₃ ,), 2.84-2.89 (m, 1H, H-2 'a), 2.70 (s, 3H, 4'-OMe), 2.44 (s, 3H, 6'-CH ₃ ), 2.38-2.42 (m, 1H, H-2'b); ¹³ C NMR (150 MHz , DMSO- D ₆ ) Δ 172.0, 167.3, 154.0,138.8, 136.3, 132.7, 131.6, 129.3, 125.9,125.8, 125.2, 125.0,123.8, 122.7, 120.7, 119.6, 119.5, 119.5 , 115.2, 114.2, 113.6, 112.8, 112.1, 109.1, 106.7, 100.8, 94.8, 82.5, 82.4, 60.2, 55.4, 53.3, 45.5,34.5 , 29.3, ^27.1 ; ( _calcd for _C40H35N6O4S , _{695.2435 )} .

Example 5 Preparation of compound 5

Under argon protection, in a 100 mL two-necked reaction flask, add tryptamine (200 mg, 1.25 mmol), dissolve it in 15 mL of tetrahydrofuran, add 0.5 mL of triethylamine, add di-tert-butyl dicarbonate (301 mg, 1.38 mmol), reacted at 10 °C for 1 h, added water to terminate the reaction, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, separated by silica gel column chromatography, eluted with petroleum ether: ethyl acetate (v/v 3:1) to obtain a white powder Product tert-butyl N- [2-(3-indole)ethyl]carbamate (250 mg, 77% yield), ESI-MS m/z 261.3 [M + H] ⁺ . In a 100 mL two-necked reaction flask, add tert-butyl N- [2-(3-indole)ethyl]carbamate (250 mg, 0.96 mmol) and dissolve it with 11 mL of THF/H ₂ O (10:1), DDQ (436 mg, 1.92 mmol) was added at 0 °C, and the reaction was carried out at this temperature for 2 hours. The reaction solution was poured into 100 mL of ethyl acetate, washed with saturated sodium bicarbonate solution until colorless, and the ethyl acetate layer was washed with anhydrous sulfuric acid. After drying over sodium, it was concentrated, separated by silica gel column chromatography, and eluted with petroleum ether:ethyl acetate (v/v 1:1) to obtain N- [2-oxyidene-2-(3-indole)ethyl]carbamic acid tert-Butyl ester (170 mg, 64% yield), ESI-MS m/z 297.0 [M + Na] ⁺ . N- [2-Oxyidene-2-(3-bromoindole)ethyl] tert-butyl carbamate (170 mg, 0.62 mmol) was dissolved in 5 mL of trifluoroacetic acid, reacted at 10 °C for one hour, and added benzene (5 mL × 3 times) azeotropically remove the trifluoroacetic acid to obtain 158 mg of N- [2-oxyidene-2-(3-indole)ethyl]ammonium trifluoroacetate with a yield of 93%. Under argon protection, in a 100 mL two-necked reaction flask, add 2.33 g of staurosporine (5.0 mmol), dissolve it in 50 mL of tetrahydrofuran, add 2 mL of triethylamine, and add 1.3 g of di-tert-butyl dicarbonate (6.0 mmol), reacted at 0 °C for 30 min, added water to terminate the reaction, extracted with dichloromethane, dried over anhydrous sodium sulfate and concentrated, separated by silica gel column chromatography, and eluted with dichloromethane: methanol (v/v 30:1) to obtain 3'- 2.18 g of N -tert-butoxycarbonyl staurosporine, with a yield of 76%. ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 9.32 (d, J = 7.9 Hz, 1H, ArH), 8.62 (s, 1H, NH), 7.99-8.08 (m, 2H, ArH), 7.64(d , J = 8.2 Hz, 1H, ArH), 7.46-7.51 (m, 2H, ArH), 7.29-7.37 (m, 2H, ArH), 7.00 (t, J = 7.6 Hz, 1H, H-1'), 5.01 (s, 2H, H-7), 4.48-4.65 (m, 1H, H-3'), 4.28 (s, 1H, H-4'), 2.72-2.83 (m, 4H, 3'-NCH ₃ , H-2'a), 2.64 (s, 3H, 4'-OCH ₃ ), 2.34 (s, 3H, 6'-CH ₃ ), 2.14-2.21 (m, 1H, H-2'b), 1.57 /1.46 (s, 9H, 3×4''-CH ₃ ); ¹³ C NMR (125 MHz, DMSO- d ₆ ) δ 171.9, 154.9/154.2, 139.0/138.8, 136.2, 132.7,129.1, 125.7, 125.3, 124.9(2×C), 123.7, 122.6, 121.4, 120.3, 119.5, 119.4,115.2, 114.1, 113.7, 108.9, 94.6, 84.0/83.3, 82.2, 79.6/79.3, 60.5, 50.5/49.7, 45.4, 30.1, 29.5 , 28.1(3×C), 27.0; ESI-MS m/z 589.2[M + Na] ⁺ . Under argon protection, in a 100 mL two-necked reaction flask, 3'- N -tert-butoxycarbonyl staurosporine (2.0 g, 3.54 mmol) was dissolved in 30 mL of dichloromethane/methanol (1:1), at room temperature Chlorosuccinimide (745 mg, 5.6 mmol) was added, the reaction was carried out at this temperature for 6 hours, the reaction was terminated by adding water, extracted with dichloromethane, dried over anhydrous Na ₂ SO ₄ , evaporated to dryness in vacuo and separated by silica gel column chromatography , dichloromethane:methanol (v/v 30:1) to give 3-chloro-3′- N -tert-butoxycarbonylstaurosporine (1.03 g, 48% yield). ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 9.32 (d, J = 2.0 Hz, 1H, ArH), 8.70 (s, 1H, NH), 8.01-8.10 (m, 2H, ArH), 7.70 (d , J = 8.0 Hz, 1H, ArH), 7.47-7.53 (m, 2H, ArH), 7.35 (t, J = 8.0 Hz, 1H, ArH), 7.01 (t, J = 7.6 Hz, 1H, H-1 '), 5.01 (s, 2H, H-7), 4.44-4.62 (m, 1H, H-3'), 4.27 (brs, 1H, H-4'), 2.76 (s, 3H, 3'-NCH ₃ ), 2.60-2.68 (m, 4H, 4'-OCH ₃ , H-2'a), 2.33 (s, 3H, 6'-CH ₃ ), 2.11-2.20 (m,1H, H-2'b ), 1.56/1.45 (m, 9H, 3×4''-CH ₃ ); ¹³ C NMR (100 MHz, DMSO- d ₆ ) δ 171.8, 154.9/154.2, 138.8, 134.6, 133.2, 128.9, 125.7, 125.2 , 125.1, 124.6, 123.8,123.7, 123.6, 121.6, 120.5, 119.4, 114.6, 114.2, 113.9/113.8, 110.7, 94.7,83.9/83.2, 82.3, 79.7/79.4, 60.5, 50.4/49.6, 45.6, 30.2, 29.5 , 28.1(3×C), 27.0; ESI-MS m/z 623.3[M + Na] ⁺ . In a 100 mL two-necked reaction flask, add 3-chloro-3′- N -tert-butoxycarbonyl staurosporine (1.03 g, 1.72 mmol), and dissolve with 10 mL of dichloromethane. 8 mL of trifluoroacetic acid was added at 0°C, reacted at this temperature for 1 hour, concentrated, added with 50 mL of saturated aqueous sodium bicarbonate solution, stirred for 0.5 hour, extracted with dichloromethane, dried over anhydrous sodium sulfate, concentrated, and separated by silica gel column chromatography. Dichloromethane:ethyl acetate (v/v 5:1) eluted to give 3-chlorostaurosporine (800 mg, 93% yield). ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 9.32 (d, J = 2.1 Hz, 1H, ArH), 8.61 (s, 1H, NH), 7.96-7.99 (m, 2H, ArH), 7.64 (d , J = 8.0 Hz, 1H, ArH), 7.46(dd, J = 8.0, 2.0 Hz, 1H, ArH), 7.42 (t, J = 8.0 Hz, 1H, ArH), 7.28 (t, J =8.0 Hz, 1H, ArH), 6.69-6.71 (m, 1H, H-1'), 4.96 (s, 2H, H-7), 4.05 (d, J =3.5 Hz, 1H, H-3'), 3.85-3.87 (m, 1H, H-4'), 3.34 (s, 3H, 4'-OCH ₃ ), 3.23-3.26(m, 1H, H-3'), 2.47-2.52 (m, 1H, H-2' a), 2.28-2.32 (m, 4H, H-2'b, 3'-NCH ₃ ), 1.42 (s, 3H, 6'-CH ₃ ); ¹³ C NMR (150 MHz, DMSO- d ₆ ) δ 172.1, 139.5, 134.7, 132.5,129.9, 127.6, 124.5 (2 × c), 124.4, 123.7, 123.3, 120.9, 119.8, 118.7, 113.9, 110.0, 91.1, 80.0, 57.2, 49.8, 49.8 , 45.5, 33.2, 29.7, 29.3; ESI-MS m/z 523.2[M + Na] ⁺ . 3-Chlorostaurosporine (470 mg, 0.94 mmol) was dissolved in 20 mL of dichloromethane, 2 mL of triethylamine and N,N'-thiocarbonyldiimidazole (502 mg, 2.82 mmol) were added at room temperature, overnight at room temperature reaction. The reaction solution was poured into 20 mL of ice water, extracted with dichloromethane, the organic phase was dried over anhydrous sodium sulfate and then concentrated, separated by silica gel column chromatography, eluted with dichloromethane:methanol=30:1 (v/v) to obtain 3-chloro-3 '-N-(1 - imidazolethiocarbonyl)staurosporine 410 mg, yield 71%. ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 9.34 (s, 1H, ArH), 8.71 (s, 1H, NH), 8.11 (s, 1H, ArH), 8.07 (d, J = 7.8 Hz, 1H , ArH), 8.03 (d, J = 8.3 Hz, 1H, ArH), 7.51-7.68 (m, 4H, ArH), 7.38 (t, J = 7.4 Hz, 1H, ArH), 7.14 (brs, 1H, ArH) ), 7.05 (brs, 1H, H-1'), 5.46 (brs, 1H, H-3'), 5.01 (s, 2H, H-7), 4.78 (brs, 1H, H-4'), 3.03 -3.07 (m, 4H, 3'-NCH ₃ , H-2'a), 2.70 (s, 3H, 4'-OMe), 2.39-2.46 (m,4H, 6'-CH ₃ , H-2'b); ¹³ C NMR (150 MHz, DMSO- d ₆ ) δ 179.4, 171.8, 138.8, 137.7, 134.6, 133.1, 129.1, 129.0, 126.0, 125.5, 125.2, 120.7, 123.9, 2.7, 123.7, 123.9, 2.7, 123.8, 12 119.9, 119.6, 114.8, 114.3, 113.5, 110.6, 94.9, 82.0, 81.7, 60.4, 58.2, 45.6, 38.2, 29.4, 27.0; ESI-MS m/z 633.0[M +Na] ⁺ . 3-Chloro-3'- N- (1-imidazolylthiocarbonyl)staurosporine (360 mg, 0.59 mmol) was dissolved in 10 mL of acetonitrile, 0.5 mL of iodomethane was added, and the reaction was carried out at room temperature for 24 hours. The reaction solution was directly concentrated, washed with a mixed solution of 50 mL petroleum ether: dichloromethane=4:1 (v/v) to obtain 3-chloro-3'- N- (1-imidazolethiocarbonyl)staurosporine Iodomethane salt of imidazole moiety 300 mg, yield 68%. ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 9.69 (s, 1H, ArH), 9.35 (d, J = 1.8 Hz, 1H, ArH), 8.73 (s, 1H, NH), 8.14 (brs, 1H) , ArH), 8.10 (d, J = 7.7 Hz, 1H, ArH), 8.07 (d, J =8.3 Hz, 1H, ArH), 7.87 (brs, 1H, ArH), 7.69 (d, J = 8.0 Hz, 1H, ArH), 7.52-7.55 (m, 2H, ArH), 7.39 (t, J = 7.5 Hz, 1H, ArH), 7.19 (brs, 1H, H-1'), 5.45(d, J = 11.3 Hz , 1H, H-3'), 5.02 (s, 2H, H-7), 4.75 (s, 1H, H-4'), 3.90 (s,3H, 5''-NCH ₃ ), 3.05-3.11 ( m, 4H, 3'-NCH ₃ , H-2'a), 2.69 (s, 3H, 4'-OMe), 2.43-2.48 (m, 4H, 6'-CH ₃ , H-2'b); ¹³ C NMR (150 MHz, DMSO- d ₆ ) δ 174.3, 171.7, 138.7, 138.0, 134.6, 133.1, 129.1, 126.0, 125.5, 125.2, 124.7, 123.9, 123.8, 123.7(2×C), 120.7, 119.6, 114.8, 114.3, 113.5, 110.6, 94.7, 81.9,81.2, 60.5, 59.1, 45.6, 38.5, 36.5, 29.2, 26.7; ESI-MS m/z 625.3[MI] ⁺ 3-Chloro-3'- N- (1-imidazolylthiocarbonyl) iodomethane salt of the imidazole moiety of staurosporine (280 mg, 0.37 mmol) was dissolved in 10 mL of DMF, triethylamine 1.0 mL and N- [ 2-Oxyidene-2-(3-indole)ethyl]ammonium trifluoroacetate (200 mg, 0.74 mmol, ) was reacted at room temperature for 24 hours. The reaction solution was diluted with 20 mL of ethyl acetate, washed with 1N hydrochloric acid, dried over anhydrous sodium sulfate and concentrated. Silica gel column chromatography, petroleum ether:ethyl acetate=1:1 (v/v) elution to obtain 3-chloro-3'- N- [ N- (2-oxyidene-2-(3-indole) Ethyl)carbamoyl]staurosporine 130 mg, yield 49%. ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 12.03 (s, 1H, NH), 9.33 (d, J = 2.0 Hz, 1H, ArH), 8.70 (s, 1H, NH), 8.50 (d, J = 1.7 Hz, 1H, ArH), 8.18 (d, J = 7.3 Hz, 1H, ArH), 8.07 (d, J = 7.8 Hz, 1H, ArH), 8.02 (d, J = 8.5 Hz, 1H, ArH) , 7.92 (brs, 1H, NH), 7.79(d, J = 8.7 Hz, 1H, ArH), 7.49-7.53 (m, 3H, ArH), 7.37 (t, J = 7.4 Hz, 1H, ArH), 7.20 -7.25 (m, 2H, ArH), 7.10 (t, J = 7.7 Hz, 1H, H-1'), 5.92 (brs, 1H,H-3'), 4.97-5.08 (m, 4H, H-7 , H-3''), 4.52 (brs, 1H, H-4'), 2.96 (s, 3H, 3'-NCH ₃ ), 2.85 (s, 3H, 4'-OCH ₃ ), 2.71-2.76 ( m, 1H, H-2'a), 2.36 (s, 3H, 6'-CH ₃ ), 2.26-2.32 (m, 1H, H-2'b); ¹³ C NMR (150MHz, DMSO- d ₆ ) δ 190.0, 182.4, 171.8,139.1, 136.4, 134.7, 133.3, 133.2, 128.9, 125.8, 125.4, 125.3, 125.1, 124.5,123.8, 123.7(2×C), 122.9, 121.8, 121.5, 121.2, 120.5, 119.4, 114.7, 114.2, 114.1, 113.9, 112.2, 110.9, 95.1, 82.8, 82.5, 60.4, 54.2, 51.9, 45.6, 32.8, 29.5, 27.6; ESI-MS m/z 739.2[M + Na] ⁺ . Under argon protection, in a 25 mL two-necked reaction flask, add 3-chloro-3'- N- [ N- (2-oxyidene-2-(3-indole)ethyl)carbamoyl] Staurosporine (61 mg, 0.085 mmol) was dissolved in 6 mL of dichloromethane and 200 µL of ethanol, and 400 µL of trifluoroacetic anhydride was added at 0°C. After 1 hour of reaction, water was added to terminate the reaction, and extracted with dichloromethane. It was dried over sodium sulfate, concentrated, separated by semi-preparative HPLC, and eluted with MeOH:H ₂ O=9:1 (v/v) to obtain 43 mg of 3-chlorofred carbazole base A (compound 5), with a yield of 73%. . ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 11.36 (s, 1H, NH), 9.35 (s, 1H, ArH), 8.71 (s, 1H, NH), 8.07 (d, J = 7.8 Hz, 1H , ArH), 8.04 (d, J = 8.6 Hz, 1H, ArH), 7.82 (d, J = 7.8 Hz, 1H, ArH), 7.2(d, J = 7.8 Hz, 1H, ArH), 7.58 (brs, 1H, ArH), 7.55 (brs, 1H, ArH), 7.52 (dd, J = 8.7, 2.1 Hz, 1H, ArH), 7.49 (t, J = 7.8 Hz, 1H, ArH), 7.45 (d, J = 8.1Hz, 1H, ArH), 7.37 (t, J = 7.4 Hz, 1H, ArH), 7.18 (t, J = 7.5 Hz, 1H, ArH), 7.08-7.13 (m, 2H , ArH, H-1' ), 5.02 (s, 2H, H-7), 4.97 (d, J = 12.0 Hz, 1H,H-3'), 4.49 (brs, 1H, H-4'), 2.91 (s, 3H, 3' -NCH ₃ ), 2.84-2.90 (m, 1H, H-2'a), 2.70 (s, 3H, 4'-OCH ₃ ), 2.43 (s, 3H, 6'-CH ₃ ), 2.37-2.42 ( m, 1H, H-2'b); ¹³ C NMR (150 MHz, DMSO- d ₆ ) δ 171.8, 167.4, 138.8, 136.5, 134.7, 133.9, 133.1, 129.1, 125.9, 125.3, 125.1, 124.8, 124.6, 123.8, 123.7(2×C), 122.8, 121.8, 121.6,120.6, 120.5, 119.7, 119.4, 119.2, 114.7, 114.2, 113.6, 112.0, 110.8, 106.9,94,6, 0.2, 5(2×C) 45.6, 34.5, 29.2, 27.0; HRESI-MS m/z 699.1938[M + H] ⁺ (calcd for C ₃₉ H ₃₂ N ₆ O ₃ ClS, 699.19 40).

Example 6 Preparation of compound 6

3-Chloro-3'- N- (1-imidazolylthiocarbonyl) iodomethane salt of the imidazole moiety of staurosporine (300 mg, 0.40 mmol) was dissolved in 10 mL of DMF, triethylamine 1.0 mL and N- [2-Oxyidene-2-(3-(5-fluoroindole))ethyl]ammonium trifluoroacetate (116 mg, 0.40 mmol) was reacted at room temperature for 24 hours. The reaction solution was diluted with 20 mL of ethyl acetate, washed with 1N hydrochloric acid, dried over anhydrous sodium sulfate and concentrated. Semi-preparative HPLC separation, MeOH: _H2O =9:1 (v/v) eluting to give 3-chloro-3'- N- [ N- (2-oxyidene-2-(3-(5-fluoro Indole))ethyl)carbamoyl]staurosporine 150 mg, 50% yield. ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 12.16 (s, 1H, NH), 9.33 (d, J = 2.1 Hz, 1H, ArH), 8.71 (s, 1H, NH), 8.58 (s, 1H , ArH), 8.07 (d, J = 7.7 Hz, 1H, ArH), 8.02 (d, J = 8.5 Hz, 1H, ArH), 7.95 (brs, 1H, NH), 7.85 (dd, J = 9.8, 2.1 Hz, 1H, ArH), 7.80 (d, J = 8.7 Hz, 1H, ArH), 7.50-7.54 (m, 3H, ArH), 7.38 (t, J =7.4 Hz, 1H, ArH), 7.09-7.12 ( m, 2H, ArH, H-1'), 5.92 (brs, 1H, H-3'), 4.95-5.05 (m, 4H, H-7, H-3''), 4.52 (brs, 1H, H -4'), 2.95 (s, 3H, 3'-NCH ₃ ), 2.86(s, 3H, 4'-OCH ₃ ), 2.72-2.76 (m, 1H, H-2'a), 2.36 (s, 3H, 6'-CH ₃ ), 2.26-2.32(m, 1H, H-2'b); ¹³ C NMR (150 MHz, DMSO- d ₆ ) δ 190.1, 182.5, 171.8, 158.6 (d, ¹ J _CF = 234.1 Hz), 139.1, 134.8 (2×C), 133.2, 133.0, 128.9, 126.0 (d, ³ J _CF =10.0 Hz), 125.8, 125.3, 125.1, 124.5, 123.8, 123.7(2×C), 121.6 , 120.5, 119.4, 114.7, 114.3 (d, ⁴ J _CF = 4.0 Hz), 114.3, 114.0, 113.5 (d, ³ J _CF = 10.0 Hz), 111.1 (d, ² J _CF = 23.8 Hz), 110.9, 106.0 (d, ² J _CF = 23.8 Hz), 95.1, 82.8, 82.5, 60.4, 54.3, 51.9, 45.6, 32.8, 29.5, 27.6; ESI-MS m/z 757.2 [M + Na] ⁺ . Under argon protection, in a 25 mL two-necked reaction flask, add 3-chloro-3'- N- [ N- (2-oxyidene-2-(3-(5-fluoroindole))ethyl)amino Thioformyl ]staurosporine (50 mg, 0.068 mmol) was dissolved in 6 mL of dichloromethane and 200 µL of ethanol, 400 µL of trifluoroacetic anhydride was added at 0°C, and the reaction was terminated by adding water for 1 hour. Chloromethane extraction, dried over anhydrous sodium sulfate and concentrated, semi-preparative HPLC separation, MeOH:H ₂ O=9:1 (v/v) elution to obtain 3-chloro-5'''-fluorofried carbazole base Formazan (Compound 6) 30 mg, yield 62%. ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 11.47 (s, 1H, NH), 9.34 (s, 1H, ArH), 8.71 (s, 1H, NH), 8.02-8.08 (m, 2H, ArH) , 7.72 (t, J = 7.9 Hz, 1H, ArH), 7.65 (brs, 1H, ArH), 7.43-7.56 (m, 5H, ArH), 7.35-7.38 (m, 1H, ArH), 7.07-7.10 ( m, 1H, ArH), 7.03 (t, J = 9.0 Hz, 1H, H-1'), 5.02 (s, 2H, H-7), 4.96 (d, J = 12.7 Hz, 1H, H-3' ), 4.49 (brs, 1H, H-4'), 2.90 (s, 3H, 3'-NCH ₃ ), 2.85-2.89 (m, 1H, H-2'a), 2.70 (s, 3H, 4' -OCH ₃ ), 2.43 (s, 3H, 6'-CH ₃ ), 2.37-2.41 (m, 1H, H-2'b); ¹³ C NMR (150 MHz, DMSO- d ₆ ) δ 171.8, 167.6, 157.4 (d, ¹ J _CF = 232.8 Hz), 138.8, 134.7, 134.1, 133.2, 133.1, 129.1, 126.0, 125.3, 125.2, 124.9, 124.9 (d, ³ J _CF = 9.7 Hz), 124.6, 123.8 C), 123.7, 121.6, 120.6, 120.0, 119.4, 114.7, 114.2, 113.7, 113.1 (d, ³ J _CF = 9.7 Hz), 110.8, 110.1(d, ² J _CF = 26.2 Hz), 107.2 (d, ⁴ J _CF = 4.4 Hz), 104.0 (d, ² J _CF = 26.2 Hz), 94.9, 82.5, 82.5, 60.2, 53.2, 45.6, 34.5, 29.2, 27.0; HRESI-MS m/z 717.1843[M + H] ⁺ ( _calcd for _{C39H31N6O3ClFS} , _{717.1845 )} .

Example 7 Preparation of compound 7

The iodomethane salt of the imidazole moiety of 3-chloro-3'- N- (1-imidazolethiocarbonyl)staurosporine (110 mg, 0.15 mmol) was dissolved in 10 mL of DMF, triethylamine 1.0 mL and N- [2-Oxyidene-2-(3-(5-chloroindole))ethyl]ammonium trifluoroacetate (46 mg, 0.15 mmol,) was reacted at room temperature for 24 hours. The reaction solution was diluted with 20 mL of ethyl acetate, washed with 1N hydrochloric acid, dried over anhydrous sodium sulfate and concentrated. Semi-preparative HPLC separation, MeOH: _H2O =9:1 (v/v) eluting to give 3-chloro-3'- N- [ N- (2-oxyidene-2-(3-(5-chloro Indole))ethyl)carbamoyl]staurosporine 50 mg, yield 45%. ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 9.32 (d, J = 2.0 Hz, 1H, ArH), 8.71 (s, 1H, NH), 8.58 (s, 1H, ArH), 8.16 (d, J = 2.0 Hz, 1H, ArH), 8.07 (d, J = 7.8 Hz, 1H, ArH), 8.02 (d, J = 8.5 Hz, 1H, ArH), 7.96 (brs, 1H, NH), 7.79 (d, J = 8.7Hz, 1H, ArH), 7.49-7.55 (m, 3H, ArH), 7.37 (t, J = 7.4 Hz, 1H, ArH), 7.26 (dd, J = 8.6, 2.1 Hz, 1H, ArH) , 7.10 (t, J = 7.7 Hz, 1H,H-1'), 5.91 (brs, 1H, H-3'), 4.94-5.07 (m, 4H, H-7, H-3''), 4.52 (brs, 1H, H-4'), 2.95 (s, 3H, 3'-NCH ₃ ), 2.85 (s, 3H, 4'-OCH ₃ ), 2.72-2.76 (m, 1H, H-2'a ), 2.36 (s, 3H, 6'-CH ₃ ), 2.26-2.32 (m, 1H, H-2'b); ¹³ C NMR (150 MHz, DMSO- d ₆ ) δ 190.2, 182.5, 171.8, 139.1 , 135.0, 134.7, 133.2, 128.9, 126.6, 126.5, 125.7, 125.1, 124.5,123.8, 123.7 (2 × c), 122.9, 121.7, 120.5, 120.3, 114.2,114.0, 113.9, 113.9 113.8, 110.9, 95.1, 82.8, 82.5, 60.4, 54.2, 51.9, 45.6, 32.8, 29.5, 27.6; ESI-MS m/z 773.0[M+Na] ⁺ . Under the protection of argon, in a 25 mL two-necked reaction flask, add 3-chloro-3'- N- [ N- (2-oxyidene-2-(3-(5-chloroindole))ethyl)amino Thioformyl ]staurosporine (27 mg, 0.036 mmol) was dissolved in 6 mL of dichloromethane and 200 µL of ethanol, 400 µL of trifluoroacetic anhydride was added at 0°C, and the reaction was terminated by adding water after 1 hour of reaction. Extracted with methyl chloride, dried over anhydrous sodium sulfate, concentrated, separated by semi-preparative HPLC, eluted with MeOH:H ₂ O=9:1 (v/v) to obtain 3,5'''-dichlorofried carbazole base methyl (Compound 7) 14 mg, yield 53%. ¹ H NMR (600MHz, DMSO- d ₆ ) δ 11.70 (s, 1H, NH), 9.35 (s, 1H, ArH), 8.71 (s, 1H, NH), 8.08 (d, J = 7.7 Hz, 1H, ArH), 8.05 (d, J = 8.4 Hz, 1H, ArH), 7.78 (brs, 1H, ArH), 7.73 (d, J = 8.6 Hz, 1H, ArH), 7.66 (brs, 1H, ArH), 7.45 -7.56 (m, 4H, ArH), 7.37 (t, J = 7.0 Hz, 1H, ArH), 7.18 (d, J = 8.6 Hz, 1H, ArH), 7.10 (t, J =7.0 Hz, 1H, H -1'), 5.03 (s, 2H, H-7), 4.98 (d, J = 13.0 Hz, 1H, H-3'), 4.49(brs, 1H, H-4'), 2.91 (s, 3H , 3'-NCH ₃ ), 2.86-2.90 (m, 1H, H-2'a), 2.70 (s, 3H, 4'-OCH ₃ ), 2.42 (s, 3H, 6'-CH ₃ ), 2.38 -2.42 (m, 1H, H-2'b); ¹³ C NMR (150MHz, DMSO- d ₆ ) δ 171.8, 167.7, 138.8, 135.0, 134.7, 134.4, 133.1, 129.1, 128.5, 125.9 (2×C) , 125.3, 125.1, 124.7, 124.6, 124.3, 123.8, 123.6,121.8, 121.6, 120.5, 119.6, 118.2, 114.7, 113.6 (2 × c), 106.7,94.9, 82.5, 60.2, 60.2, 60.2 53.1, 45.6, 34.5, 29.2, 27.0; HRESI-MS m/z _755.1368 [ _M +H] ⁺ ( _calcd for _{C39H30N6O3Cl2NaS} , _755.1369 ).

Example 8 Preparation of Compound 8

The iodomethane salt of the imidazole moiety of 3-chloro-3'- N- (1-imidazolethiocarbonyl)staurosporine (110 mg, 0.15 mmol) was dissolved in 10 mL of DMF, triethylamine 1.0 mL and N- [2-Oxyidene-2-(3-(5-bromoindole))ethyl]ammonium trifluoroacetate (52 mg, 0.15 mmol,) was reacted at room temperature for 24 hours. The reaction solution was diluted with 20 mL of ethyl acetate, washed with 1N hydrochloric acid, dried over anhydrous sodium sulfate and concentrated. Semi-preparative HPLC separation, eluting with MeOH: _H2O =9:1 (v/v) gave 3-chloro-3'- N- [ N- (2-oxyidene-2-(3-(5-bromo Indole))ethyl)carbamoyl]staurosporine 45 mg, 38% yield. ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 12.22 (s, 1H, NH), 9.33 (s, 1H, ArH), 8.70 (s, 1H, NH), 8.57 (s, 1H, ArH), 8.31 (s, 1H, ArH), 8.07 (d, J = 8.0 Hz, 1H, ArH), 8.02 (d, J = 8.6 Hz, 1H, ArH), 7.95 (brs, 1H, NH), 7.79 (d, J = 8.6 Hz, 1H,ArH), 7.36-7.52 (m, 5H, ArH), 7.10 (t, J = 7.5 Hz, 1H, H-1'), 5.90 (brs, 1H,H-3'), 4.92 -5.05 (m, 4H, H-7, H-3''), 4.52 (brs, 1H, H-4'), 2.95 (s, 3H, 3'-NCH ₃ ), 2.86 (s, 3H, 4 '-OCH ₃ ), 2.71-2.74 (m, 1H, H-2'a), 2.35 (s, 3H, 6'-CH ₃ ), 2.26-2.31 (m, 1H, H-2'b); ¹³ C NMR (150 MHz, DMSO- d ₆ ) δ 190.2, 182.5, 171.8, 139.1, 135.1, 134.7, 134.5, 133.2, 128.9, 127.2, 125.7, 125.5, 125.3, 125.1, 123.5, 23.7(×) , 123.3, 121.5, 120.5, 119.4, 114.7, 114.6, 114.3,114.2, 114.0, 113.6, 110.9, 95.1, 82.5, 60.4, 54.2, 45.6, 32.8,29.5, 27.6; ESI-MS M/ Z817.0.0 [M + Na] ⁺ . Under the protection of argon, in a 25 mL two-necked reaction flask, add 3-chloro-3'- N- [ N- (2-oxyidene-2-(3-(5-bromoindole))ethyl)amino Thioformyl ]staurosporine (30 mg, 0.038 mmol) was dissolved in 6 mL of dichloromethane and 200 µL of ethanol, and 400 µL of trifluoroacetic anhydride was added at 0°C. After 1 hour of reaction, water was added to terminate the reaction. Extracted with methyl chloride, dried over anhydrous sodium sulfate, concentrated, separated by semi-preparative HPLC, eluted with MeOH:H ₂ O=9:1 (v/v) to obtain 3-chloro-5'''-bromofried carbazole base A (compound 8) 14 mg, yield 47%. ¹ H NMR (600MHz, DMSO- d ₆ ) δ 11.58 (s, 1H, NH), 9.35 (d, J = 1.8 Hz, 1H, ArH), 8.71 (s, 1H, NH), 8.08 (d, J = 7.8 Hz, 1H, ArH), 8.04 (d, J = 8.5 Hz, 1H, ArH), 7.92(brs, 1H, ArH), 7.73 (d, J = 8.7 Hz, 1H, ArH), 7.65 (d, J = 2.4 Hz, 1H, ArH), 7.56 (brs, 1H, ArH), 7.48-7.54 (m, 2H, ArH), 7.42 (d, J = 8.6 Hz, 1H, ArH), 7.37 (t, J = 7.4 Hz, 1H, ArH), 7.29 (dd, J = 8.6, 1.8 Hz, 1H, ArH), 7.09-7.11(m, 1H, H-1'), 5.03 (s, 2H, H-7), 4.96- 4.99 (m, 1H, H-3'), 4.49 (s, 1H, H-4'), 2.91 (s, 3H, 3'-NCH ₃ ), 2.86-2.90 (m, 1H, H-2'a ), 2.70 (s, 3H, 4'-OCH ₃ ), 2.43 (s, 3H, 6'-CH ₃ ), 2.38-2.42 (m, 1H, H-2'b); ¹³ C NMR (150 MHz, DMSO- d ₆ ) δ 171.8, 167.8, 138.8, 135.2, 134.7, 134.5, 133.1, 129.1, 126.6, 125.9, 125.3,125.1, 124.5(2×C), 124.4, 123.8, 123.7, 123.6, 121.6, 121.2, 120.5 , 119.5,119.4, 114.7, 114.2, 114.0, 113.6, 112.2, 110.8, 106.6, 94.9, 82.5, 82.4,60.2 , 53.1, 45.6 [M ⁺ 27.0; ( _calcd for _{C39H31N6O3BrClS} , _{777.1045 )} .

Example 9 Preparation of compound 9

The iodomethane salt of the imidazole moiety of 3-chloro-3'- N- (1-imidazolethiocarbonyl)staurosporine (280 mg, 0.37 mmol) was dissolved in 10 mL of DMF, triethylamine 1.0 mL and N- [2-Oxyidene-2-(3-(5-methoxyindole))ethyl]ammonium trifluoroacetate (111 mg, 0.37 mmol,) was reacted at room temperature for 24 hours. The reaction solution was diluted with 20 mL of ethyl acetate, washed with 1N hydrochloric acid, dried over anhydrous sodium sulfate and concentrated. Semi-preparative HPLC separation, MeOH: _H2O =9:1 (v/v) elution to give 3-chloro-3'- N- [ N- (2-oxyidene-2-(3-(5-methyl) Oxyindole))ethyl)carbamoyl]staurosporine 119 mg, yield 43%. ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 11.92 (s, 1H, NH), 9.33 (s, 1H, ArH), 8.70 (s, 1H, NH), 8.43 (s, 1H, ArH), 8.07 (d, J = 7.8 Hz, 1H, ArH), 8.02 (d, J = 8.5 Hz, 1H, ArH), 7.90 (brs, 1H, NH), 7.79 (d, J = 8.7 Hz, 1H, ArH), 7.69(s, 1H, ArH), 7.48-7.52 (m, 2H, ArH), 7.36-7.40 (m, 2H, ArH), 7.10 (t, J =7.6 Hz, 1H, H-1'), 6.87 ( dd, J = 8.7, 2.4 Hz, 1H, ArH), 5.93 (brs, 1H, H-3'), 4.95-5.03 (m, 4H, H-7, H-3''), 4.50 (s, 1H , H-4'), 3.78 (s, 3H, 5'''-OCH ₃ ), 2.95 (s, 3H, 3'-NCH ₃ ), 2.83 (s, 3H, 4'-OCH ₃ ), 2.71- 2.75 (m, 1H, H-2'a), 2.36(s, 3H, 6'-CH ₃ ), 2.26-2.32 (m, 1H, H-2'b); ¹³ C NMR (150 MHz, DMSO- D ₆ ) Δ 189.9,182.4, 171.8, 155.5, 139.1, 134.8, 133.5, 131.3, 129.0, 126.3, 125.8,125.3, 124.5, 123.7 (2 × c), 121.6, 120.5, 119.4, 114.7.7 , 114.3,114.0(2×C), 113.0, 112.7, 110.9, 102.9, 95.1, 82.9, 82.5, 60.4, 55.3, 54.2,51.8 , 45.6, 32.7, 29.4, 27.6; Na] ⁺ . Under argon protection, in a 25 mL two-necked reaction flask, add 3-chloro-3'- N- [ N- (2-oxyidene-2-(3-(5-methoxyindole))ethyl )carbamoyl]staurosporine (35 mg, 0.047 mmol), add 6 mL of dichloromethane and 200 µL of ethanol to dissolve, add 400 µL of trifluoroacetic anhydride at 0°C, and add water to terminate the reaction after 1 hour of reaction. Extracted with methyl chloride, dried over anhydrous sodium sulfate, concentrated, separated by semi-preparative HPLC, eluted with MeOH:H ₂ O=9:1 (v/v) to obtain 3-chloro-5'''-methoxyfredcarb 16 mg of azole base (compound 9), yield 47%. ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 11.20 (s, 1H, NH), 9.34 (s, 1H, ArH), 8.71 (s, 1H, NH), 8.08 (d, J = 7.8 Hz, 1H , ArH), 8.05 (d, J = 8.6 Hz, 1H,ArH), 7.72 (dd, J = 8.7, 2.0 Hz, 1H, ArH), 7.55 (s, 1H, ArH), 7.48-7.53(m,3H , ArH), 7.3-7.38 (m, 2H, ArH), 7.24 (brs, 1H, ArH), 7.08-7.11 (m, 1H, H-1'), 6.83 (dd, J = 8.8, 2.0 Hz, 1H , ArH), 5.02 (s, 2H, H-7), 4.97 (d, J =12.0 Hz, 1H, H-3'), 4.50 (s, 1H, H-4'), 3.83 (s, 3H, 5'''-OCH ₃ ), 2.90 (s, 3H,3'-NCH ₃ ), 2.86-2.89 (m, 1H, H-2'a), 2.67 (s, 3H, 4'-OCH ₃ ), 2.43 (s, 3H, 6'-CH ₃ ), 2.38-2.42 (m, 1H, H-2'b); ¹³ C NMR (150 MHz, DMSO- d ₆ ) δ 171.8, 167.3, 153.9, 138.8, 134.7 , 133.7, 133.1, 131.6, 129.1, 125.9, 125.3, 125.1 (2 × c), 124.6, 123.8, 123.7, 123.5, 121.6, 120.5, 119.4, 114.2,113.6, 112.0, 110.8, 110.8, 110.8 106.7, 100.8, 94.9, 82.5(2×C), 60.2, 55.4, 53.1, 45.6, 34.5, 29.2, 27.0; HRESI-MS m/z 729.2040 [M + H] ⁺ (calcd for C ₄₀ H ₃₄ N ₆ O _4ClS , 729.2045).

Example 10 Preparation of compound 10

Under argon protection, in a 25 mL two-neck reaction flask, add 3'-N- tert-butoxycarbonyl staurosporine (1.0 g, 1.77 mmol), dissolved in 30 mL of dichloromethane/methanol (1:1), bromosuccinimide (347 mg, 1.95 mmol) was added at 0°C, The reaction was carried out at this temperature for 0.5 hours, water was added to terminate the reaction, extracted with dichloromethane, and the reaction was carried out with anhydrous Na₂SO₄ dried, evaporated to dryness in vacuo, separated by silica gel column chromatography, petroleum ether: ethyl acetate (v/v 1:1) to obtain 3-bromo-3'-N- 1.07 g of tert-butoxycarbonyl staurosporine, the yield is 94%.¹H NMR (400 MHz, DMSO-d ₆)δ 9.48 (s, 1H, ArH), 8.71 (s, 1H, NH), 8.00-8.10 (m, 2H, ArH), 7.60-7.66 (m, 2H, ArH), 7.50 (brs, 1H), 7.36(t ,J = 7.4 Hz, 1H, ArH), 7.01 (t,J = 7.5 Hz, 1H, H-1'), 5.01 (s, 2H, H-7), 4.44-4.63 (m, 1H, H-3'), 4.28 (brs, 1H, H-4'), 2.76 (s, 3H, 3'-NCH₃), 2.60-2.67 (m, 4H, 4'-OCH₃, H-2'a), 2.33 (s, 3H, 6'-CH₃), 2.11-2.20 (m, 1H, H-2’b), 1.56/1.46 (m, 9H, 3×4’’-CH₃);¹³C NMR (100 MHz, DMSO-d ₆) Δ 171.8, 154.8/154.2, 138.8, 134.9, 133.2, 128.8, 127.6, 125.5, 125.2, 124.3, 123.6,121.6, 120.4, 114.6, 113.8/113.7, 111.1.1, 94.7, 83.9/ 83.2, 82.3, 79.7/79.3, 60.5, 50.4/49.6, 45.6, 30.1, 29.5, 28.1(3×C), 26.9; ESI-MSm/z 667.2[M+Na]⁺. In a 100 mL two-necked reaction flask, add 3-bromo-3'-N- tert-Butoxycarbonyl staurosporine (1.07 g, 1.66 mmol), dissolved in 10 mL of dichloromethane. 8 mL of trifluoroacetic acid was added at 0°C, reacted at this temperature for 1 hour, concentrated, added with 50 mL of saturated aqueous sodium bicarbonate solution, stirred for 0.5 hour, extracted with dichloromethane, dried over anhydrous sodium sulfate, concentrated, and separated by silica gel column chromatography. Dichloromethane:ethyl acetate (v/v 5:1) eluted to give 3-bromostaurosporine (450 mg, 50% yield).¹H NMR (400 MHz, CDCl₃) δ 9.56 (d,J = 2.0 Hz, 1H, ArH), 7.92 (d,J = 8.0 Hz, 1H, ArH), 7.89 (d,J = 8.0 Hz, 1H, ArH), 7.54 (d,J = 8.0, 2.0Hz, 1H, ArH), 7.41-7.44 (m, 1H, ArH), 7.31-7.34 (m, 1H, ArH), 7.16 (d,J =8.0 Hz, 1H, ArH), 6.49-6.51 (m, 1H, H-1'), 6.45 (s, 1H, NH), 5.01 (s, 2H, H-7), 3.85-3.87 (m, 1H , H-4'), 3.42 (s, 3H, 4'-OCH₃), 3.33-3.35 (m, 1H, H-3'), 2.69-2.72 (m, 1H, H-2'a), 2.34-2.39 (m, 4H, H-2'b, 3'-NCH₃ ), 1.50 (s, 3H,6’-CH₃);¹³C NMR (100 MHz, CDCl₃) δ 173.3, 139.8, 135.1, 132.5, 130.6, 128.9,127.7, 127.7, 124.0, 124.4, 124.3, 120.6, 120.1, 118.4, 115.3, 114.4, 114.3,112.6, 108.3, 91.0, 84.1, 80.1, 57.2, 50.1, 45.9, 33.3, 30.1, 29.7; ESI-MSm/ z 567.1[M+Na]⁺. 3-Bromostaurosporine (600 mg, 1.1 mmol) was dissolved in 30 mL of dichloromethane, 3 mL of triethylamine and N,N'-thiocarbonyldiimidazole (587 mg, 3.3 mmol) were added at room temperature, and the room temperature was overnight reaction. The reaction solution was poured into 30 mL of ice water, extracted with dichloromethane, the organic phase was dried over anhydrous sodium sulfate and then concentrated, separated by silica gel column chromatography, and eluted with dichloromethane:methanol=30:1 (v/v) to obtain 3-bromo- 3'-N-(1-Imidazolethiocarbonyl)staurosporine 631 mg, 88% yield.¹HNMR (600 MHz, DMSO-d ₆)δ 9.45 (s, 1H, ArH), 8.71 (s, 1H, NH), 8.12 (brs, 1H, ArH), 8.08 (d,J = 7.8 Hz, 1H, ArH), 8.04 (d,J = 7.8 Hz, 1H, ArH) , 7.64(brs, 1H, ArH), 7.62-7.66 (m, 2H, ArH), 7.53 (t,J = 7.8 Hz, 1H, ArH), 7.38(t,J = 7.8 Hz, 1H, ArH), 7.15 (brs, 1H, ArH), 7.05 (brs, 1H, H-1'), 5.46(brs, 1H, H-3'), 5.03 (s, 2H, H- 7), 4.78 (brs, 1H, H-4'), 3.00-3.08 (m, 4H,3'-NCH₃, H-2’a), 2.76 (s, 3H, 4’-OMe), 2.38-2.46 (m, 4H, 6’-CH₃, H-2’b);¹³CNMR (150 MHz, DMSO-d ₆)δ 179.4, 171.7, 138.9, 137.7, 134.9, 133.2, 129.1,129.0, 127.7, 127.6, 125.8, 124.4, 121.7, 120.7, 119.6, 114.2, 111.8, 111.8, 94.9, 82.0, 82.0, 82.9 81.7, 60.4, 58.1, 45.6, 38.2, 29.4, 26.9; ESI-MSm/z 677.2[M+Na]⁺. 3-bromo-3'-N-(1-Imidazolethiocarbonyl)staurosporine (583 mg, 0.89 mmol) was dissolved in 45 mL of acetonitrile, 1 mL of iodomethane was added, and the reaction was carried out at room temperature for 24 hours. The reaction solution was directly concentrated, washed with a mixed solution of 50 mL petroleum ether: dichloromethane=4:1 (v/v) to obtain 3-bromo-3'-N-(1-Imidazolethiocarbonyl) Iodomethane salt of the imidazole moiety of staurosporine 521 mg, yield 74%.¹H NMR (600 MHz, DMSO-d ₆) δ 9.69(s, 1H, ArH), 9.50 (s, 1H, ArH), 8.71 (s, 1H, NH), 8.14 (brs, 1H, ArH), 8.09(d,J = 7.7 Hz, 1H, ArH), 8.06 (d,J = 8.4 Hz, 1H, ArH), 7.86 (brs, 1H, ArH), 7.63-7.67 (m, 2H, ArH), 7.54 (t,J = 7.6 Hz, 1H, ArH), 7.39 (t,J = 7.6 Hz, 1H, ArH), 7.17-7.20 (m, 1H), 5.39 (d,J = 12.4 Hz, 1H, H-3’), 5.02 (s, 2H, H-7), 4.75 (s, 1H, H-4’), 3.92 (s, 3H, 5’’-NCH₃), 3.06-3.11 (m, 4H, 3’-NCH₃, H-2’a), 2.71 (s, 3H, 4’-OMe), 2.43-2.47 (m, 4H, 6’-CH₃, H-2’b);¹³C NMR (150MHz, DMSO-d ₆)δ 174.3, 171.7, 138.7, 138.0, 134.9, 133.1, 129.1, 127.7,127.6, 125.8, 124.4, 123.7, 121.8, 121.2, 119.6, 114.9, 113.5, 111.8, 94.7, 81.9, 81.9, 81.9, 81.9, 81.9, 81.9, 81.9, 81.9, 81.9, 81.9, 81.9, 81.9, 81.9, 81.9, 81.9, 81.9, 81.9, 81.9, 94.7, 81.9, 81.9, 94.7, 81.9, 81.9, 94.7, 81. 81.2, 60.5, 59.1, 45.6, 38.5, 36.5, 29.2, 26.7; ESI-MSm/z 669.2[M-I]⁺. 3-Bromo-3'-N-(1-Imidazolethiocarbonyl) staurosporine imidazole moiety methyl iodide salt (110 mg, 0.14 mmol) was dissolved in 10 mL DMF, triethylamine 1.0 mL andN-[2-Oxyidene-2-(3-indolyl)ethyl]ammonium trifluoroacetate (38 mg, 0.14 mmol,) was reacted at room temperature for 24 hours. The reaction solution was diluted with 20 mL of ethyl acetate, washed with 1N hydrochloric acid, dried over anhydrous sodium sulfate and concentrated. Semi-preparative HPLC separation, MeOH:H₂O=9:1 (v/v) eluted to give 3-bromo-3'-N-[N-(2-Oxyidene-2-(3-indole)ethyl)carbamoyl]staurosporine 52 mg, yield 49%.¹H NMR (600 MHz, DMSO-d ₆)δ 12.03 (d,J = 2.4 Hz, 1H, NH), 9.48(d,J = 2.0 Hz, 1H, ArH), 8.70 (s, 1H, NH), 8.50 (d,J = 3.0 Hz, 1H, ArH), 8.18 (d,J= 7.5 Hz, 1H, ArH), 8.07 (d,J = 7.8 Hz, 1H, ArH), 8.02 (d,J =8.5 Hz, 1H, ArH), 7.92 (brs, 1H, NH), 7.75 (d,J = 8.7 Hz, 1H, ArH), 7.63(dd,J = 8.6, 2.0 Hz, 1H, ArH), 7.49-7.52 (m, 2H, ArH), 7.37 (t,J = 7.4 Hz, 1H, ArH), 7.19-7.25 (m, 2H, ArH), 7.09 (t,J = 8.0 Hz, 1H, H-1'), 5.92 (brs,1H, H-3'), 4.97-5.07 (m, 4H, H-7, H-3'), 4.52 (s, 1H, H- 4'), 2.95 (s, 3H, 3'-NCH₃), 2.85 (s, 3H, 4'-OCH₃), 2.71-2.75 (m, 1H, H-2'a), 2.36 (s, 3H, 6'-CH₃), 2.26-2.31 (m, 1H, H-2’b);¹³C NMR (150 MHz, DMSO-d6)δ190.0, 182.4, 171.8,139.1, 136.4, 135.0, 133.3, 133.2, 128.9, 127.7, 125.6, 125.4, 125.2,124.3, 122.9, 121.6, 120.5, 119.7, 114.7, 2 × 2 × C), 114.0, 112.2, 111.6, 111.3, 95.1, 82.8, 82.5, 60.4, 54.2, 51.9, 45.6, 32.6, 29.5, 27.5; ESI-MSm/z 783.2[M+Na]⁺. Under the protection of argon, in a 25 mL two-necked reaction flask, add 3-bromo-3'-N-[N-(2-Oxyidene-2-(3-indole)ethyl)carbamoyl]staurosporine (36 mg, 0.047 mmol), dissolved in 6 mL of dichloromethane and 200 µL of ethanol, 0°C 400 µL of trifluoroacetic anhydride was added to the bottom of the reaction mixture, and water was added to terminate the reaction after 1 hour of reaction, extracted with dichloromethane, dried over anhydrous sodium sulfate and concentrated, separated by semi-preparative HPLC, MeOH:H₂O=9:1 (v/v) eluted to obtain 19 mg of 3-bromofried carbazole base A (compound 10) with a yield of 55%.¹H NMR (600 MHz, DMSO-d ₆)δ11.38 (s, 1H, NH), 9.50 (s, 1H, ArH), 8.71 (s, 1H, NH), 8.07 (d,J = 7.6 Hz, 1H), 8.04 (d,J = 8.3 Hz, 1H), 7.82 (d,J = 7.7 Hz, 1H, ArH), 7.62-7.69 (m,2H, ArH), 7.58 (s, 1H, ArH), 7.55 (s, 1H, ArH), 7.49 (t,J = 7.7 Hz, 1H, ArH), 7.45 (d,J = 8.0 Hz, 1H, ArH), 7.37 (t,J= 7.2 Hz, 1H, ArH), 7.18 (t,J= 7.4 Hz, 1H, ArH), 7.08-7.13 (m, 2H, ArH, H-1’), 5.03 (s, 2H, H-7), 4.97(d,J = 12.8 Hz, 1H), 4.49 (s, 1H, H-4'), 2.91 (s, 3H, 3'-NCH₃), 2.85-2.89(m, 1H, H-2'a), 2.70 (s, 3H, 4'-OCH₃), 2.43 (s, 3H, 6'-CH₃), 2.37-2.42 (m, 1H,H-2’b);¹³C NMR (150 MHz, DMSO-d ₆)δ 171.8, 167.4, 138.8, 136.5, 135.0, 133.9,133.1, 129.0, 127.7, 125.8, 125.3, 124.8, 123.6, 121.8,121.6, 120.6, 119.6, 119.1, 114.7, 114.0, 114.0, 114.7, 114.0, 114.0, 114.0, 114.0, 114.0, 114.0, 114.0, 114.0, 114.0, 114.0, 114.7, 114.0, 114.7, 114.0, 114.7, 114.0, 114.7, 114.0, 114.7, 114.0, 114.7, 114.7, 114.7, 114.7, 114.7, 114.0, 114.7, 114.7, 114.0, 114.7, 114.0, 113.6, 112.0, 111.6, 111.2, 106.9, 94.9, 82.5, 82.5, 60.2, 53.1, 45.6, 34.5, 29.2, 27.0; HRESI-MS m/z 743.1431 [M+H]⁺ (calcd for C₃₉H₃₂N₆O₃BrS, 743.1434).

Example 11 Preparation of compound 11

3-Bromo-3'-N-(1-imidazolethiocarbonyl) staurosporine imidazole moiety methyl iodide salt (228 mg, 0.29 mmol) was dissolved in 10 mL of DMF, triethylamine 1.0 mL andN-[2-Oxyidene-2-(3-(5-fluoroindole))ethyl]ammonium trifluoroacetate (84 mg, 0.29 mmol,) was reacted at room temperature for 24 hours. The reaction solution was diluted with 20 mL of ethyl acetate, washed with 1N hydrochloric acid, dried over anhydrous sodium sulfate and concentrated. Semi-preparative HPLC separation, MeOH:H₂O=9:1 (v/v) eluted to give 3-bromo-3'-N-[N-(2-Oxyidene-2-(3-(5-fluoroindole))ethyl)carbamoyl]staurosporine 101 mg, 45% yield.¹H NMR (600 MHz, DMSO-d ₆)δ12.16 (s, 1H, NH), 9.48 (s, 1H, ArH), 8.70 (s, 1H, NH), 8.57 (s, 1H, ArH), 8.07 (d,J = 7.8 Hz, 1H, ArH), 8.01 (d,J = 8.5 Hz, 1H, ArH), 7.94 (s, 1H, NH), 7.85 (d,J = 9.8 Hz, 1H, ArH), 7.74 (d,J = 8.7Hz, 1H, ArH), 7.62 (dd,J = 8.6, 2.0 Hz, 1H, ArH), 7.48-7.54 (m, 2H, ArH), 7.37 (t,J = 7.4 Hz, 1H, ArH), 7.07-7.11 (m, 2H, ArH, H-1'), 5.91 (brs, 1H,H-3'), 4.95-5.05 (m, 4H, H-7, H -3''), 4.51 (brs, 1H, H-4'), 2.95 (s, 3H, 3'-NCH₃), 2.84 (s, 3H, 4'-OCH₃), 2.71-2.75 (m, 1H, H-2'a), 2.36 (s, 3H, 6'-CH₃), 2.26-2.33 (m, 1H, H-2’b);¹³C NMR (150 MHz, DMSO-d ₆)δ190.2, 182.5, 171.8, 158.6 (d,¹ J _CF = 234.8 Hz), 139.1, 135.0, 134.9, 133.3, 133.1, 128.9, 127.7, 127.6, 126.1 (d,³ J _CF = 10.8 Hz), 125.6, 125.3, 124.4, 123.7, 121.6, 120.5, 119.4, 114.8, 114.3 (d,⁴ J _CF = 4.2 Hz), 114.2, 114.0, 113.5 (d,³ J _CF = 10.8Hz), 111.7, 111.4, 111.1 (d,² J _CF = 24.0 Hz), 106.0 (d,² J _CF = 24.0 Hz), 95.1, 82.9, 82.5, 60.4, 54.3, 51.9, 45.7, 32.8, 29.5, 27.6; ESI-MSm/z 801.2 [M+Na]⁺. Under the protection of argon, in a 25 mL two-necked reaction flask, add 3-bromo-3'-N-[N-(2-Oxylidene-2-(3-(5-fluoroindole))ethyl)aminothioyl]staurosporine (50 mg, 0.064 mmol), 6 mL of dichloromethane and 200µDissolve in L ethanol, add 400µL trifluoroacetic anhydride, reacted for 1 hour, added water to terminate the reaction, extracted with dichloromethane, dried over anhydrous sodium sulfate, concentrated, separated by semi-preparative HPLC, MeOH:H₂O=9:1 (v/v) eluted to obtain 24 mg of 3-bromo-5'''-fluorofred carbazole base A (compound 11) with a yield of 50%.¹H NMR (600 MHz, DMSO-d ₆)δ11.49 (s, 1H, NH), 9.49 (s, 1H, ArH), 8.71 (s, 1H, NH), 8.07 (d,J = 7.8 Hz, 1H, ArH), 8.03 (d,J = 8.5 Hz, 1H, ArH), 7.62-7.68 (m, 3H, ArH), 7.53-7.56 (m, 2H, ArH), 7.48(t,J = 7.8 Hz, 1H, ArH), 7.44-7.46 (m, 1H, ArH), 7.36 (t,J = 7.4 Hz, 1H,ArH), 7.01-7.10 (m, 2H, ArH, H-1’), 5.02 (s, 2H, H-7), 4.96 (d,J = 12.0 Hz, 1H, H-3'), 4.48 (brs, 1H, H-4'), 2.88 (s, 3H, 3'-NCH₃), 2.84-2.88 (m, 1H, H-2'a), 2.69 (s, 3H, 4'-OCH₃), 2.43 (s, 3H, 6'-CH₃), 2.37-2.41 (m, 1H, H-2’b); ¹³C NMR (150 MHz, DMSO-d ₆)δ171.8, 167.5, 157.4 (d,¹ J _CF = 231.8 Hz), 138.8, 135.0, 134.1, 133.2, 133.1, 129.0, 127.7, 127.6, 125.8, 125.3, 124.9 (d,³ J _CF= 9.8 Hz), 124.9, 124.3, 123.6, 121.6, 120.5, 120.0, 119.4, 114.7, 114.1, 113.6, 113.1 (d,³ J _CF = 9.8 Hz), 111.7, 111.2, 110.1(d,² J _CF = 25.4 Hz), 107.2(d,⁴ J _CF = 4.5 Hz), 103.9 (d,² J _CF = 25.4 Hz), 94.9, 82.5, 82.4, 60.2, 53.1, 45.6, 34.5, 29.2, 27.0; HRESI-MSm/z 761.1337 [M+H]⁺ (calcd for C₃₉H₃₁N₆O₃BrFS, 761.1340).

Example 12 Preparation of compound 12

3-Bromo-3'- N- (1-imidazolylthiocarbonyl) iodomethane salt of the imidazole moiety of staurosporine (200 mg, 0.25 mmol) was dissolved in 10 mL of DMF, triethylamine 1.0 mL and N- [2-Oxyidene-2-(3-(5-chloroindole))ethyl]ammonium trifluoroacetate (76 mg, 0.25 mmol,) was reacted at room temperature for 24 hours. The reaction solution was diluted with 20 mL of ethyl acetate, washed with 1N hydrochloric acid, dried over anhydrous sodium sulfate and concentrated. Semi-preparative HPLC separation, eluting with MeOH: _H2O =9:1 (v/v) gave 3-bromo-3'- N- [ N- (2-oxyidene-2-(3-(5-chloro Indole))ethyl)carbamoyl]staurosporine 90 mg, 44% yield. ¹ HNMR (600 MHz, DMSO- d ₆ ) δ 12.21 (s, 1H, NH), 9.47 (d, J = 2.0 Hz, 1H, ArH), 8.70 (s, 1H, NH), 8.58 (s, 1H, ArH), 8.16 (d, J = 2.0 Hz, 1H, ArH), 8.07 (d, J = 7.8 Hz, 1H, ArH), 8.02 (d, J = 8.5 Hz, 1H, ArH), 7.95 (brs, 1H , NH), 7.75(d, J = 8.7 Hz, 1H, ArH), 7.63 (dd, J = 8.6, 2.0 Hz, 1H, ArH), 7.54 (d, J =8.6 Hz, 1H), 7.51 (t, J = 7.7 Hz, 1H), 7.37 (t, J = 7.4 Hz, 1H, ArH), 7.26(dd, J = 8.6, 2.1 Hz, 1H, ArH), 7.09 (t, J = 7.7 Hz, 1H, H) -1'), 5.90 (brs,1H, H-3'), 4.93-5.05 (m, 4H, H-7, H-3'), 4.52 (s, 1H, H-4'), 2.95 (s , 3H, 3'-NCH ₃ ), 2.86 (s, 3H, 4'-OCH ₃ ), 2.71-2.75 (m, 1H, H-2'a), 2.35 (s, 3H, 6'-CH ₃ ) , 2.26-2.32 (m, 1H, H-2'b); ¹³ C NMR (150 MHz, DMSO- d ₆ ) δ 190.2, 182.5, 171.8, 139.1, 135.0, 134.9, 134.7, 133.2, 128.9, 127.7, 127.5 , 126.6, 126.5, 1256,125.3, 124.3, 123.7, 122.9, 121.6, 120.5, 119.4, 114.7, 114.0, 113.9, 111.6, 111.3, 95.1, 82.8, 60.4, 54.9, 45.6, 45.6, 45.6, 45.6 , 32.8, 29.5, 27.5; ESI-MS m/z 817.0[M + Na] ⁺ . Under the protection of argon, in a 25 mL two-necked reaction flask, add 3-bromo-3'- N- [ N- (2-oxyidene-2-(3-(5-chloroindole))ethyl)amino Thioformyl]staurosporine (38 mg, 0.048 mmol) was dissolved in 6 mL of dichloromethane and 200 µL of ethanol, and 400 µL of trifluoroacetic anhydride was added at 0°C. After 1 hour of reaction, water was added to terminate the reaction. Dichloromethane Extraction, drying over anhydrous sodium sulfate, concentration, semi-preparative HPLC separation, MeOH:H ₂ O=9:1 (v/v) elution to obtain 3-bromo-5'''-chlorofried carbazole base methyl ( Compound 12) 18 mg, yield 48%. ¹ H NMR (600MHz, DMSO- d ₆ ) δ 11.58 (s, 1H, NH), 9.50 (s, 1H, ArH), 8.71 (s, 1H, NH), 8.06 (t, J = 7.5 Hz, 1H, ArH), 8.03 (d, J = 8.4 Hz, 1H, ArH), 7.79 (brs, 1H, ArH), 7.62-7.68 (m, 3H, ArH), 7.56 (s, 1H, ArH), 7.46-7.50 ( m, 2H, ArH), 7.36 (t, J = 7.5 Hz, 1H, ArH), 7.18 (dd, J = 8.4, 1.9 Hz, 1H, ArH), 7.09 (brs, 1H, H-1'), 5.02 (s, 2H, H-7), 4.95-4.99 (m, 1H, H-3'), 4.48 (s, 1H, H-4'), 2.90 (s,3H, 3'-NCH ₃ ), 2.85 -2.89 (m, 1H, H-2'a), 2.69 (s, 3H, 4'-OCH ₃ ), 2.43 (s, 3H,6'-CH ₃ ), 2.37-2.42 (m, 1H, H- 2'b); ¹³ C NMR (150 MHz, DMSO- d ₆ ) δ 171.8, 167.7, 138.8, 134.9 (2×C), 134.5, 133.1, 129.0, 127.7, 127.6, 125.9, 125.8, 125.3, 124.7, 12 (2×C), 123.6, 121.9, 121.6, 120.5, 119.6, 119.4, 118.2, 114.7, 114.0, 113.6 , 29.2, 27.0; HRESI-MS m/z _777.1044 [ _M +H] ⁺ (calcd for _{C39H31N6O3BrClS} , _777.1045 ).

Example 13 Preparation of compound 13

3-Bromo-3'- N- (1-imidazolylthiocarbonyl) iodomethane salt of the imidazole moiety of staurosporine (100 mg, 0.13 mmol) was dissolved in 10 mL of DMF, triethylamine 1.0 mL and N- [2-Oxyidene-2-(3-(5-chloroindole))ethyl]ammonium trifluoroacetate (45 mg, 0.13 mmol) was reacted at room temperature for 24 hours. The reaction solution was diluted with 20 mL of ethyl acetate, washed with 1N hydrochloric acid, dried over anhydrous sodium sulfate and concentrated. Semi-preparative HPLC separation, eluting with MeOH: _H2O =9:1 (v/v) gave 3-bromo-3'- N- [ N- (2-oxyidene-2-(3-(5-bromo Indole))ethyl)carbamoyl]staurosporine 45 mg, 42% yield. ¹ HNMR (600 MHz, DMSO- d ₆ ) δ 9.48 (d, J = 1.8 Hz, 1H, ArH), 8.71 (s, 1H, NH), 8.56 (s, 1H, ArH), 8.31 (d, J = 1.6 Hz, 1H, ArH), 8.07 (d, J = 7.8 Hz, 1H, ArH), 8.02 (d, J = 8.5 Hz, 1H, ArH), 7.97 (brs, 1H, NH), 7.75 (d, J = 8.7 Hz, 1H, ArH), 7.63 (dd, J = 8.6, 1.8 Hz, 1H, ArH), 7.49-7.52 (m, 2H, ArH), 7.36-7.38 (m, 2H, ArH), 7.09 (t , J = 7.7 Hz, 1H, H-1'), 5.90 (s, 1H, H-3'), 4.92-5.07 (m, 4H, H-7, H-3'), 4.52 (s, 1H, H-4'), 2.95 (s, 3H, 3'-NCH ₃ ), 2.86 (s, 3H, 4'-OCH ₃ ), 2.71-2.74 (m, 1H, H-2'a), 2.35 (s , 3H, 6'-CH ₃ ), 2.26-2.31 (m,1H, H-2'b); ¹³ C NMR (150 MHz, DMSO- d ₆ ) δ 190.2, 182.5, 171.8, 139.1, 135.3,135.0, 134.6, 133.2, 127.7, 127.5, 127.3, 125.6, 125.4, 124.3,123.7, 121.6, 120.5, 119.4, 114.5, 114.1, 113.6, 111.3, 111.3, 95.1, 82.8, 82.8, 82.8, 82.8, 82.8, 82.8, 82.8, 82.8, 82.8, 82.8, 82.8.8, 82.8, 82.8, 82.8.8. 82.5, 60.4, 54.2, 51.9, 45.6, 32.8, 29.5, 27.5; ESI-MS m/z 861.2 [M + Na] ⁺ . Under argon protection, in a 25 mL two-necked reaction flask, add 3-bromo-3'- N- [ N- (2-oxyidene-2-(3-(5-bromoindole))ethyl)amino Thioformyl ]staurosporine (50 mg, 0.060 mmol) was dissolved in 6 mL of dichloromethane and 200 µL of ethanol, 400 µL of trifluoroacetic anhydride was added at 0°C, and the reaction was terminated by adding water for 1 hour. Extracted with methyl chloride, dried over anhydrous sodium sulfate, concentrated, separated by semi-preparative HPLC, eluted with MeOH:H ₂ O=9:1 (v/v) to obtain 3,5'''-dibromofried carbazole base methyl (Compound 13) 26 mg, yield 53%. ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 11.66 (s, 1H, NH), 9.49 (s, 1H, ArH), 8.71 (s, 1H, NH), 8.07 (d, J = 7.7 Hz, 1H , ArH), 8.04 (d, J = 8.5 Hz, 1H, ArH), 7.92 (s, 1H, ArH), 7.62-7.69 (m, 3H, ArH), 7.55 (s, 1H, ArH), 7.49 (d , J = 8.0 Hz, 1H, ArH), 7.43 (d, J = 8.5 Hz, 1H, ArH), 7.37 (t, J = 7.4 Hz, 1H, ArH), 7.28-7.30 (m, 1H, ArH), 7.08-7.11(m, 1H, H-1'), 5.04 (s, 2H, H-7), 4.97 (d, J = 12.7 Hz, 1H, H-3'), 4.49 (s,1H, H- 4'), 2.91 (s, 3H, 3'-NCH ₃ ), 2.85-2.89 (m, 1H, H-2'a), 2.69 (s, 3H, 4'-OCH ₃ ), 2.43 (s, 3H , 6'-CH ₃ ), 2.36-2.41 (m, 1H, H-2'b); ¹³ C NMR (150 MHz, DMSO- d ₆ ) δ 171.8, 167.8, 138.8, 135.2, 135.0, 134.5, 133.1, 129.1, 127.7, 127.6,126.6, 125.8, 125.3, 124.5, 124.4, 124.3, 123.6, 121.6, 121.2, 120.5, 119.5,119.4, 114.7, 114.1(2×C), 113.6, 112.2, 111.7, 111.2, 106.6, 94.9 , 82.5, 82.4, 60.2, 53.1, 45.6, 34.5, 29.2, 27.0; HRESI-MS m/z 821.0539 [M + H] ⁺ (calcd for C ₃₉ H ₃₁ N ₆ O ₃ Br ₂ S, 821.0539).

Example 14 Preparation of compound 14

3-Bromo-3'- N- (1-imidazolylthiocarbonyl) iodomethane salt of the imidazole moiety of staurosporine (200 mg, 0.25 mmol) was dissolved in 10 mL of DMF, triethylamine 1.0 mL and N- [2-Oxyidene-2-(3-(5-methoxyindole))ethyl]ammonium trifluoroacetate (75 mg, 0.25 mmol) was reacted at room temperature for 24 hours. The reaction solution was diluted with 20 mL of ethyl acetate, washed with 1N hydrochloric acid, dried over anhydrous sodium sulfate and concentrated. Semi-preparative HPLC separation, MeOH: _H2O =9:1 (v/v) eluting to give 3-bromo-3'- N- [ N- (2-oxyidene-2-(3-(5-methyl) Oxyindole))ethyl)carbamoyl]staurosporine 109 mg, yield 56%. ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 11.92 (s, 1H, NH), 9.47 (s, 1H, ArH), 8.70 (s, 1H, NH), 8.43 (d, J = 2.0 Hz, 1H , ArH), 8.07 (d, J = 7.6 Hz, 1H, ArH), 8.02 (d, J = 8.4 Hz, 1H, ArH), 7.89 (brs, 1H, NH), 7.74 (d, J = 8.6 Hz, 1H,ArH), 7.69 (s, 1H, ArH), 7.63 (d, J = 8.6 Hz, 1H, ArH), 7.50 (t, J = 7.6 Hz, 1H, ArH), 7.36-7.41 (m, 2H, ArH), 7.09 (t, J = 7.6 Hz, 1H, H-1'), 6.87 (dd, J = 8.6, 2.1 Hz, 1H, ArH), 5.93 (brs, 1H, H-3'), 4.93- 5.03 (m, 4H, H-7, H-3''), 4.50 (s, 1H, H-4'), 3.78 (s, 3H, 5'''-OCH ₃ ), 2.95 (s, 3H, 3'-NCH ₃ ), 2.82 (s,3H, 4'-OCH ₃ ), 2.71-2.75 (m, 1H, H-2'a), 2.36 (s, 3H, 6'-CH ₃ ), 2.26- 2.32 (m,1H, H-2'b); ¹³ C NMR (150 MHz, DMSO- d ₆ ) δ 189.9, 182.4, 171.8, 155.5, 139.1, 135.0, 133.4, 133.2, 131.3, 129.0, 127.7, 127.5, 126.3, 125.6, 125.3, 124.3,123.7, 121.6, 120.5, 119.4, 114.7, 114.1, 113.9 (2×C), 113.0, 112.7, 111.6,111.4, 102.9, 95.1, 82.9, 82.5, 60.4, 55.3, 54.2, 51.8 , 45.6, 32.7, 29.4, 27.6; ESI-MS m/z 813.2[M + Na] ⁺ . Under argon protection, in a 25 mL two-necked reaction flask, add 3-bromo-3'- N- [ N- (2-oxyidene-2-(3-(5-methoxyindole))ethyl )carbamoyl]staurosporine (56 mg, 0.071mmol), add 6 mL of dichloromethane and 200 µL of ethanol to dissolve, add 400 µL of trifluoroacetic anhydride at 0°C, and add water to terminate the reaction after 1 hour of reaction. Extracted with methyl chloride, dried over anhydrous sodium sulfate, concentrated, separated by semi-preparative HPLC, eluted with MeOH:H ₂ O=9:1 (v/v) to obtain 3-bromo-5'''-methoxyfredcarb azole base A (compound 14) 32 mg, yield 59%. ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 11.21 (s, 1H, NH), 9.50 (s, 1H, ArH), 8.71 (s, 1H, NH), 8.07 (d, J = 7.8 Hz, 1H , ArH), 8.04 (d, J = 8.5 Hz, 1H, ArH), 7.62-7.68 (m, 2H, ArH), 7.55 (s, 1H, ArH), 7.51 (s, 1H, ArH), 7.49 (t , J = 7.8 Hz, 1H,ArH), 7.34-7.37 (m, 2H, ArH), 7.24 (s, 1H, ArH), 7.09 (t, J = 7.6 Hz, 1H, H-1'), 6.83 ( dd, J = 8.5, 1.6 Hz, 1H), 5.04 (s, 2H, H-7), 4.96 (d, J = 12.0 Hz, 1H, H-3'), 4.49 (s, 1H, H-4' ), 3.83 (s, 3H, 5'''-OCH ₃ ), 2.90 (s, 3H, 3'-NCH ₃ ), 2.85-2.89 (m, 1H, H-2'a), 2.68 (s, 3H , 4'-OCH ₃ ), 2.43 (s, 3H, 6'-CH ₃ ), 2.37-2.43 (m, 1H, H-2'b); ¹³ C NMR (150MHz, DMSO- d ₆ ) δ 171.8, 167.3, 153.9, 133.7, 133.7, 133.1, 131.6, 127.7, 127.6, 125.8, 125.1, 124.4,123.6, 121.6, 120.5, 119.7, 114.1, 113.6, 112.0, 112.0, 112.0, 112.7 ^111.7 , 111.2 , _106.7 , _100.8 , 94.9, 82.5, 82.5, 60.2, 55.4, 53.1, 45.6, 34.5, 29.1, 27.0 _; O ₄ BrS, 773.1540).

In order to further verify the beneficial effects of the compounds synthesized in the present invention, the compounds synthesized in the schemes of Examples 1-14 were tested for antitumor activity, and the specific experiments were as follows:

1. Experimental method

Preparation of the tested sample solution: The test samples are compounds 1-14 synthesized in the above-mentioned Examples 1-14. Accurately weigh an appropriate amount of sample and prepare a solution of the required concentration with DMSO for the activity test.

Cell lines and subculture of cells: K562, MV-4-11, HL-60 and PBMC cells were used for viability assays. Various cells were subcultured in RPMI-1640 medium containing 10% FBS at 37°C in an incubator with 5% carbon dioxide.

In this experiment, Cell Titer Glo (CTG) method was used to detect the growth inhibitory effects of different drugs on K562, MV-4-11, HL-60 and PBMC cells.

CTG method: Adenosine triphosphate (referred to as adenosine triphosphate, ATP) participates in various enzymatic reactions in the body, and is an indicator of the metabolism of living cells, and its content directly reflects the number and state of cells. During the experiment, an equal volume of CTG reagent was added to the cell culture medium, and the luminescence value was measured. In the light signal and system, the luminescence value is proportional to the amount of ATP, and ATP is positively correlated with the number of living cells. Therefore, cells can be obtained by detecting the content of ATP. vitality.

During the activity test, K562, MV-4-11, HL-60 and PBMC cells in logarithmic growth phase were taken and prepared into a cell suspension with a density of 2 × 10 ⁴ cells per ml with fresh IMDM medium. After counting, it was inoculated into a 96-well culture plate, 2000 cells/100 μL /well, 90 μL of culture medium (containing serum) was added to each well and cultured overnight, then 10 μL of drug solution was added, and the culture was continued for 48 hours. Then 100 μL CTG was added, and it was allowed to stand at room temperature for 10 min, and the luminescence value was measured, 0.5 ms. Calculate the cell proliferation rate (%) at each concentration according to the following formula: IR% = (chemiluminescence value of control group - chemiluminescence value of experimental group)/chemiluminescence value of control group × 100%. Using Graph Pad software, _IC50 was calculated.

PKC-412 was the positive control group.

2. Experimental results

The experimental results are shown in Table 1.

Table 1. Inhibitory activity (IC ₅₀ ) of fred carbazole base A derivatives 1~14 on human leukemia cell proliferation

compound MV4-11 (<i>μ</i>M) K562 (<i>μ</i>M) HL-60 (<i>μ</i>M) PBMC (<i>μ</i>M) 1 0.513 >10 >10 >10 2 0.422 >10 >10 >10 3 0.413 >10 >10 >10 4 0.364 >10 >10 >10 5 0.564 >10 >10 >10 6 0.321 >10 >10 >10 7 0.589 >10 >10 >10 8 0.426 >10 >10 >10 9 0.441 >10 >10 >10 10 0.355 >10 >10 >10 11 0.964 >10 >10 >10 12 0.698 >10 >10 >10 13 0.394 >10 >10 >10 14 0.512 >10 >10 >10 PKC-412 0.037 >10 >10 >10

It can be seen from Table 1 that

(1) Compounds 1-14 have strong inhibitory activity against human leukemia cell line MV4-11 , with IC ₅₀ below 1 μM.

(2) Compounds 1-14 have weak inhibitory effects on PBMC of human peripheral blood mononuclear cells, with IC ₅₀ greater than 10 μM .

It shows that all the compounds of the present invention have highly selective inhibitory effect on human leukemia cell line MV4-11 (weak activity on other leukemia cell lines HL-60 and K562, IC ₅₀ is greater than 10 μM ).

3. Experimental conclusion

It can be seen from the above experiments that the synthetic fred carbazole base A compound selectively has a strong inhibitory activity on the human leukemia cell MV4-11 cell line, and has an inhibitory effect on the human peripheral blood mononuclear cell PBMC cell line. weaker, which indicates that it can be developed as a highly effective and low toxicity drug for the prevention and treatment of leukemia.

The above-mentioned embodiments are merely examples for clear illustration, and are not intended to limit the implementation manner. For those of ordinary skill in the art, changes or modifications in other different forms can also be made on the basis of the above description. There is no need and cannot be exhaustive of all implementations here. And the obvious changes or changes derived from this are still within the protection scope of the present invention.

Claims (2)

1. a Fried carbazole base class A compound, it is characterized in that: this compound is made on the basis of chlorostaurosporine, and its structural formula is formula (I):

2. The application of a Friedcarbazole base A compound according to claim 1 in the preparation of a medicine for preventing or treating leukemia.

Images