Preparation and application of anti-leukemia silyl-narcotine derivative

Abstract

The invention belongs to the technical field of medicines, and relates to synthesis of a brand-new silicon-substituted narcotine derivative and application of the brand-new silicon-substituted narcotine derivative in the anti-leukemia field. The invention tries to introduce a silicon group and a silyl group to the 9'site of narcotine to synthesize the silicon-substituted narcotine derivative with a novel structure, and the silicon-substituted narcotine derivative prepared by the invention shows more excellent proliferation inhibition activity on leukemia cells than narcotine, can obviously induce leukemia cell apoptosis, and has a good development prospect.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology and relates to the synthesis of a new type of silanacrotine derivatives and their application in the field of anti-leukemia. Background Art

[0002] Leukemia is a group of malignant tumors of the hematopoietic system that originates from abnormally proliferating leukemic cells in the bone marrow, interfering with normal blood cell production. Based on the course of the disease, it is classified as acute (rapidly progressive) or chronic (slowly progressive), and based on the cell type, it is divided into lymphocytic and myeloid leukemias. Acute lymphoblastic leukemia (ALL) accounts for approximately 75% of childhood leukemias, of which the T-cell subtype (T-ALL) accounts for 10%-15%. In adults, acute myeloid leukemia (AML) is more common, accounting for 80% of acute leukemias. Common symptoms in patients include anemia, bleeding, infection, and organ infiltration. Some types, such as T-ALL, are prone to mediastinal masses and central nervous system involvement.

[0003] Currently, the treatment of leukemia is mainly based on chemotherapy, combined with targeted drugs and hematopoietic stem cell transplantation (HSCT). For example, the use of tyrosine kinase inhibitors significantly improves the prognosis of patients with Ph chromosome-positive ALL, while patients with AML may benefit from FLT3 or IDH inhibitors. Immunotherapy such as CD19 CAR-T cell therapy has made breakthroughs in B-ALL, but its effectiveness in treating T-cell malignancies such as T-ALL is limited due to target selection and damage to normal T cells. The prevention and treatment of central nervous system leukemia relies on intrathecal chemotherapy and radiotherapy, but there are problems with neurotoxicity and insufficient efficacy.

[0004] Small molecule drugs such as vincristine, cytarabine, and methotrexate are common chemotherapy agents for leukemia. However, these drugs lack tissue selectivity and suffer from severe side effects, which restricts their further application and poses a challenge to leukemia treatment. Tumor heterogeneity and the emergence of drug-resistant clones further contribute to poor prognosis for leukemia patients. Therefore, the development of highly effective and low-toxic anti-leukemia drugs is urgently needed.

[0005] Natural products are an important source of anti-tumor drugs. Noscapine is a phthalide tetrahydroisoquinoline alkaloid derived from poppy ( Figure 1), with its simple structure, abundant content, and low price, it was first widely used clinically as an over-the-counter cough suppressant and can be taken orally. At the end of the last century, it was discovered to inhibit the activity of various malignant tumor cells and has also entered clinical research as a treatment for chronic lymphocytic leukemia and Hodgkin's lymphoma. Research has shown that its anti-tumor mechanisms are diverse, with microtubule inhibitors being the most important. They bind to tubulin and disrupt microtubule homeostasis, causing tumor cells to arrest in the G2 / M phase of mitosis and undergo apoptosis, thereby exerting anti-tumor effects. However, as a microtubule inhibitor, noscapine does not alter the overall mass of tubulin, but rather its kinetic properties. Therefore, it has no significant toxic side effects and can effectively overcome the widespread drug resistance of this type of anti-tumor drug, representing a promising anti-tumor drug lead.

[0006] Organosilicon, which is generally non-toxic and has good biomembrane permeability, has unique advantages in the development of new drugs. For example, the introduction of trimethylsilylethyl and tert-butyldimethylsilyl groups at the 7-position of camptothecin has resulted in organosilicon compounds that not only have excellent anti-tumor activity but also have better oral bioavailability and metabolic stability. As a candidate drug for the treatment of glioblastoma, it has entered the clinical research stage ( Figure 2 ). Summary of the Invention

[0007] In order to develop more effective and low-toxic anti-leukemia drugs, the present invention attempts to introduce silicon and silyl groups at the 9′ position of narcotine, and synthesizes a class of novel silyl-generation narcotine derivatives, which show higher in vitro anti-leukemia activity than narcotine and are expected to be developed into new leukemia treatment drugs with high efficiency and low toxicity.

[0008] The first aspect of the present invention discloses a silanacrotine derivative, the general structural formula of which is as follows:

[0009]

[0010] R in the general structural formula Ⅰ and Ⅱ 1 and R 3 The group is selected from alkyl, deuterated alkyl; R 2 The group is selected from alkyl, deuterated alkyl, cycloalkyl, deuterated cycloalkyl, and aryl;

[0011] or

[0012]

[0013] R in the general structural formula III and IV 4 and R 5 The group is selected from alkyl and deuterated alkane, and n is any number from 1 to 6. Preferably, n is 1, 2 or 3.

[0014] As a preferred embodiment of the present invention, the structural formula of the specific compound is as follows:

[0015]

[0016]

[0017]

[0018] The second aspect of the present invention discloses a method for preparing the above-mentioned specific compounds S1-S13.

[0019] The preparation method of S1 is as follows:

[0020]

[0021] 1) Using liquid bromine as the bromination reagent, the 9′ position of Narcotine is brominated in the presence of hydrobromic acid to obtain the intermediate 9′-bromonarcotine II;

[0022] 2) Under the action of methyllithium, hexamethyldisilane generates trimethylsilyllithium intermediate, which then reacts with isopropyl borate to form trimethylsilylpinacol borate III;

[0023] 3) 9′-Bronacotine II and trimethylsilylpinacol borate undergo a coupling reaction in the presence of tetrakistriphenylphosphine palladium and potassium carbonate as a base to produce compound S1.

[0024] The preparation method of S2–S8 is as follows:

[0025]

[0026] 1) Hydrosilane reacts with biboronic acid pinacol ester under the catalysis of palladium on carbon (Pt / C) to generate silyl pinacol borate esters IV-XII;

[0027] 2) Using potassium carbonate as a base, 9′-bromonacotine II and silylpinacol borate esters IV-XII undergo coupling reaction under the catalysis of tetrakistriphenylphosphine palladium to produce compounds S2-S8.

[0028] The preparation method of S9-S11 is as follows:

[0029]

[0030] Using nickel iodide as catalyst, 4,4′-di-tert-butylbipyridine as ligand, zinc powder as reducing agent, magnesium chloride and pyridine as additives, 9′-bromonacotine II and trimethylsilyl bromide underwent Negishi coupling to produce compounds S9-S11.

[0031] The preparation method of S12 is as follows:

[0032]

[0033] 1) Narcotine I is oxidized by m-chloroperbenzoic acid (m-CPBA) and ferrous sulfate (Fe2SO4)-mediated reductive demethylation to generate intermediate XIII;

[0034] 2) Intermediate XII reacts with triphosgene (BTC) under alkaline conditions, and then reacts with dimethylamine to obtain intermediate XIV;

[0035] 3) XIV undergoes bromination reaction with N-bromosuccinimide in acetic acid to obtain intermediate XV;

[0036] 4) Using nickel iodide as a catalyst, 4,4′-di-tert-butylbipyridine as a ligand, zinc powder as a reducing agent, magnesium chloride and pyridine as additives, XV and trimethylsilyl bromide undergo Negishi coupling to produce compound S12.

[0037] The preparation method of S13 is as follows:

[0038]

[0039] Using potassium carbonate as a base, compound XV and compound IV undergo a coupling reaction catalyzed by tetrakistriphenylphosphine palladium to produce compound S13.

[0040] The third aspect of the present invention discloses the use of the above-mentioned silanacrotine derivatives in the preparation of anti-leukemia drugs.

[0041] Compared with the prior art, the present invention has the following advantages:

[0042] The silane-based narcotine derivative prepared by the present invention exhibits a more excellent proliferation inhibitory activity on leukemia cells than narcotine, and can significantly induce apoptosis of leukemia cells, thus having a good development prospect. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the existing methods and experiments, the following is a brief introduction to the drawings required for use in the embodiments. It should be understood that the following drawings only illustrate certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other relevant drawings can be obtained based on these drawings without paying any creative work.

[0044] Figure 1 This is the structural diagram of Narcotine.

[0045] Figure 2 This is the structural diagram of silicon-modified camptothecin anti-tumor drug candidate.

[0046] Figure 3This is a cell viability curve diagram of leukemia cells in Example 14 after being treated with different compounds.

[0047] Figure 4 This is a diagram showing the apoptosis of leukemia cells after being treated with different compounds in Example 14. DETAILED DESCRIPTION

[0048] The technical solutions of the present invention are described in detail below with reference to the accompanying drawings and examples, but the present invention is not limited to the scope of the examples. Experimental methods and techniques in the following examples where specific conditions are not specified are generally performed in accordance with conventional conditions in the art or the conditions recommended by the manufacturer.

[0049] Example 1: Synthesis of Compound S1

[0050] 1) Synthesis of Compound II:

[0051] Weigh 1 gram of narcotine and dissolve it in 5 mL of HBr (40% in H2O). Measure 50 mL of bromine water (3%) and add it dropwise into the narcotine hydrobromic acid solution with vigorous stirring. After the addition is complete, continue the reaction for 2 hours. After the reaction is completed, add concentrated ammonia water dropwise to quench the reaction. Adjust the pH to 9-10, add 50 mL of dichloromethane and extract three times. Combine the organic phases, dry over anhydrous sodium sulfate, filter, concentrate and spin-dry to obtain crude product II.

[0052] 2) Synthesis of Compound III

[0053] TPPA (10 mL), TMEDA (10 mL), and hexamethyldisiloxane (12.5 mmol) were injected into a nitrogen-protected Shrek tube equipped with a magnetic stirring bar, and the reaction system was cooled to -60°C. Then, an anhydrous ether solution of MeLi·LiBr (12.5 mmol) was injected into the system. After completion, the mixture was immediately heated to 20°C and stirred for 5 minutes. The mixture was then cooled to -60°C again and the reaction system was added. i PrO-Bpin (12.5 mmol) was added and stirred at 20°C for 1 hour. After the reaction, the reaction mixture was directly subjected to silica gel column chromatography (n-hexane:ethyl acetate = 30:1) as the eluent to obtain compound III as a colorless oil.

[0054] 3) Synthesis of Compound S1

[0055] Compound 9′-bronarcotine II (49.1 mg, 0.1 mmol, 1.0 equiv.), compound III (40.8 mg, 0.3 mmol, 3.0 equiv.), Pd(PPh3)4 (11.6 mg, 10 mol%), and anhydrous K2CO3 (41.4 mg, 0.3 mmol, 3.0 equiv.) were weighed and added to a dry reaction flask equipped with a magnetic stirrer. 1 mL of toluene solution was then added and mixed thoroughly. The flask was sealed with a Teflon-lined septum nut. A long needle connected to a nitrogen balloon was inserted through the septum below the liquid level. A short needle was then inserted to degas the solution (the short needle should not be inserted below the liquid level to prevent backflow). Degassing was performed by bubbling for 15 minutes. After bubbling, the reaction was sealed and placed in an oil bath at 100°C for 24 hours. After completion of the reaction, the system was cooled to room temperature, water (5 mL) was added, and the mixture was extracted three times with ethyl acetate (5 mL). The organic phases were combined. The organic phase was washed with saturated brine (10 mL), dried over anhydrous Na2SO4, concentrated under reduced pressure, and separated by column chromatography (PE:EA=2:1) ​​to give S1 (12.4 mg, 23%) as a light yellow solid. 1 HNMR(400MHz, CDCl3) δ6.89(d,J=8.0Hz,1H),5.94–5.84(m,3H),5.46(d,J=4.0Hz,1H),4.46(d,J=4.0Hz,1H),4.09(s,3H),4.06(s,3H),3.84( s,3H),2.63(dt,J=10.5,4.0Hz,1H),2.55(s,3H),2.68–2.60(m,1H),2.48–2.40(m,1H),2.28–2.20(m,1H),0.28(s,9H).HRMS(ESI)Calcd.for C 25 H 31 NO7Si[(M+H) + ]486.1943,found486.1947.

[0056] Example 2: Synthesis of Compound S2

[0057] 1) Synthesis of Compounds IV-XII

[0058] Bis(pyraclostrobin) 1 (1.25 mmol, 2.5 equiv.) and Pt / C (5 wt% Pt, 0.1 mmol) catalyst were placed in a pre-dried, stirred, two-necked flask. A condenser and a T-piece were secured, and the atmosphere was purged with nitrogen five times. Cyclohexane (5 mL) and silane (0.5 mmol, 1 equiv.) were then added and reacted in an oil bath for 20 hours. The product was then concentrated under reduced pressure and separated by column chromatography (PE:EA = 100:1–50:1) to afford the desired product.

[0059] 1) Synthesis of Compound S2 Using compound IV as starting material, according to the preparation method of Example 1, a pale yellow solid S2 (14.5 mg, 25%) was obtained. 1 HNMR(400MHz, CDCl3)δ6.87(d,J=8.0Hz,1H),5.94(d,J=8.4Hz,1H),5.90–5.83(m ,2H),5.49(d,J=4.0Hz,1H),4.09(s,3H),4.07(s,3H),3.84(s,3H),2.64–2.56(m ,1H),2.54(s,3H),2.46–2.36(m,1H),2.28–2,18(m,1H),1.78–1.58(m,6H),1.25 –1.07(m,5H),0.92–0.84(m,1H),0.26(s,3H),0.22(s,3H).HRMS(ESI)Calcd.for C 30 H 39 NO7Si[(M+H) + ]554.2569,found 554.2564.

[0060] Example 3: Synthesis of Compound S3

[0061] Using compound V as the starting material, according to the preparation method of Example 1, a light yellow solid S3 (22.8 mg, 40%) was obtained. 1 H NMR (400MHz, CDCl3) δ7.50–7.40(m,2H),7.36–7.28(m,3H),6.84(d,J=8.4Hz,2H) ,5.99(d,J=8.0Hz,1H),5.94–5.88(m,2H),5.49(d,J=4.0Hz,1H),4.44(d,J=4.0H z,1H),4.08(s,1H),4.07(s,1H),3.84(s,1H),2.48(s,1H),2.46–2.38(m,1H),2. 32–2.21(m,1H),2.15–2.05(m,1H),1.61–1.50(m,1H),0.57(s,3H),0.53(s,3H); 13C NMR (100MHz, CDCl3) δ168.3,154.8,152.3,147.8,141.7,141.2,139.7,138.5,133.7,133.0,129.1,128.0,120.5, 118.3,118.0,108.2,100.2,82.3,62.4,61.4,59.61,57.0,51.1,467.0,30.1,25.0,0.7,0.4.HRMS(ESI)Calcd.for C 30 H 34 NO7Si[(M+H) + ]548.2105,found548.2106.

[0062] Example 4: Synthesis of Compound S4

[0063] Using compound VI as the reaction raw material, according to the preparation method of Example 1, a light yellow solid S4 (9.0 mg, 17%) was obtained. 1 H NMR(400MHz, CDCl3)δ6.89(d,J=8.4Hz,1H),5.95–5.83(m,3H),5.49(d,J=4.0H z,1H),4.48(s,1H),4.09(s,3H),4.06(s,3H),3.84(s,3H),2.70–2.60(m,1H),2 .56(s,3H),2.46–2.36(m,1H),2.31–2.09(m,1H),1.80–1.67(m,1H),0.95(t,J =4.0Hz,3H),0.79–0.74(m,1H),0.28(s,3H),0.26(s,3H).HRMS(ESI)Calcd.for C 26 H 34 NO7Si[(M+H) + ]500.2099,found500.2101

[0064] Example 5: Synthesis of Compound S5

[0065] Using compound VIII as the reaction raw material, according to the preparation method of Example 1, a light yellow solid S5 (10.0 mg, 18%) was obtained. 1H NMR(400MHz, CDCl3) δ6.85(d,J=8.4Hz,1H),5.92(d,J=8.0Hz,1H),5.89–5.85(m ,2H),5.51(d,J=4.0Hz,1H),4.48(d,J=4.0Hz,1H),4.10(s,3H),4.08(s,3H),3.8 3(s,3H),2.64–2.59(m,0H),2.55(s,3H),2.48–2.38(m,1H),2.28–2.18(m,1H),1 .79–1.69(m,1H),0.94(t,J=8.0Hz,9H),0.84–0.80(m,6H).HRMS(ESI)Calcd.for C 28 H 37 NO7Si[(M+H) + ]548.2412,found 548.2413.

[0066] Example 6: Synthesis of Compound S6

[0067] Using compound IX as the reaction material, according to the preparation method of Example 1, a light yellow solid S6 (7.3 mg, 12%) was obtained. 1 H NMR(400MHz, CDCl3)δ6.90(d,J=8.4Hz,1H),6.03(s,1H),5.90–5.85(m,2H),5 .50(d,J=4.0Hz,1H),4.48(d,J=4.0Hz,1H),4.10(s,3H),4.06(s,3H),3.85(s ,3H),2.68–2.60(s,1H),2.54(s,3H),2.53–2.48(m,1H),2.23–2.15(m,1H),1 .80–1.70(m,1H),0.90(s,9H),0.34(s,3H),0.25(s,3H).HRMS(ESI)Calcd.for C 28 H 37 NO7Si[(M+H) + ]548.2412,found548.2412.

[0068] Example 7: Synthesis of Compound S7

[0069] Using compound X as the reaction material, according to the preparation method of Example 1, a light yellow solid S8 (8.1 mg, 13%) was obtained. 1H NMR(400MHz, CDCl3) δ6.86(d,J=8.4Hz,1H),5.95(d,J=8.4Hz,1H),5.88–5.85(m,2 H),5.52(d,J=4.0Hz,1H),4.47(d,J=4.0Hz,1H),4.08(s,3H),3.84(s,3H),2.65–2 .58(m,1H),2.55,(s,3H),2.48–2.40(m,1H),2.26–2.18(m,1H),1.84–1.75(m,1H) ,1.33–1.22(m,6H),0.95(t,J=7.2Hz,6H),0.82–0.77(m,6H).HRMS(ESI)Calcd.for C 31 H 43 NO7Si[(M+H) + ]570.2882,found 570.2888.

[0070] Example 8: Synthesis of Compound S8

[0071] Using compound XII as the reaction raw material and following the preparation method of Example 1, a light yellow solid S8 (5.0 mg, 6%) was obtained. 1 HNMR(400MHz, CDCl3)δ6.87(d,J=8.4Hz,1H),5.96(d,J=8.0,1H),5.87–5.85(m,2H),5 .51(d,J=4.0,Hz,1H),4.47(d,J=4.0Hz,1H),4.10(s,3H),4.08(s,3H),3.84(s,3H),2 .65–2.58(m,1H),2.54(s,3H),2.47–2.38(m,1H),2.26–2.19(m,1H),1.86–1.76(m,1H ),1.30–1.24(m,24H),0.86(t,J=6.8Hz,9H),0.81–0.76(m,6H).HRMS(ESI)Calcd.forC 40 H 61 NO7Si[(M+H) + ]696.4290,found 696.4293.

[0072] Example 9: Synthesis of Compound S9

[0073] 9'-Bronarcotine (Compound II) (49.1 mg, 0.1 mmol, 1.0 equiv.), methyl bromide (24.8 mg, 0.15 mmol, 1.5 equiv.), zinc powder (13.1 mg, 0.2 mmol, 2.0 equiv.), MgCl2 (9.5 mg, 0.1 mmol, 1.0 equiv.), pyridine (8.1 μL, 0.1 mmol, 1.0 equiv.), NiI2 (3.1 mg, 10 mol%), and dtbpy (3.6 mg, 10 mol%) were weighed into a dry reaction vial equipped with a magnetic stirrer. 1 mL of DMA solution was then added and mixed thoroughly. The reaction vial was sealed with a Teflon-lined septum nut. A long needle connected to a nitrogen balloon was inserted through the septum below the liquid level of the reaction vial. A short needle was then inserted to vent the solution (the short needle should not be inserted below the liquid level to prevent backflow). Degassing was continued by bubbling for 15 minutes. After bubbling, the reaction mixture was sealed and allowed to react at room temperature for 24 h. After completion of the reaction, the system was cooled to room temperature, water (10 mL) was added, and the mixture was extracted three times with ethyl acetate (5 mL). The organic phases were combined. The organic phases were then washed with saturated brine (10 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. Flash chromatography (PE:EA = 3:1–2:1) afforded S9 (15.6 mg, 30%) as a pale yellow solid. 1 H NMR (400MHz, CDCl3) δ6.90(d,J=8.0Hz,1H),6.07(d,J=8.0Hz,1H),5.91–5.87(m,2H),5.62(d,J=4.0Hz,1H),4.43(d,J=4.0Hz,2H),4.09(d . 13 C NMR (150MHz, CDCl3) δ168.25,152.22,147.85,145.79,141.42,137.59,133.65,129.10,120.28,118.33,117.91,1 14.02,100.40,82.07,62.47,61.31,59.59,56.91,50.22,46.38,24.32,15.96,0.25,0.80.HRMS(ESI)Calcd.forC 26 H 34 NO7Si[(M+H) +]500.2099,found 500.2106.

[0074] Example 10: Synthesis of Compound S10

[0075] Using trimethylethyl bromide as the reaction material, according to the preparation method of Example 9, a light yellow solid S10 (12.3 mg, 22%) was obtained. 1 H NMR(400MHz, CDCl3) δ6.89(d,J=8.4Hz,1H),6.00–5.90(m,3H),5.57(d,J=4.0Hz,1H),4.46(d,J=4.0Hz,1H),4.09(s,3H),3.97(s,3H), 3.84(s,3H),2.76–2.60(m,1H),2.56(s,3H),2.54–2.30(m,4H),1.8–1.70(m,1H),0.68–0.61(m,2H),0.04(s,9H).HRMS(ESI)Calcd.for C 27 H 35 NO7Si[(M+H) + ]514.2256,found 514.2257.

[0076] Example 11: Synthesis of Compound S11

[0077] Using trimethylpropane bromide as the reaction raw material, according to the preparation method of Example 9, a light yellow solid S11 (14.1 mg, 28%) was obtained. 1 H NMR(400MHz, CDCl3) δ6.89(d,J=8.4Hz,1H),6.00(d,J=8.0Hz,1H).5.96–5.90( m,2H),5.56(d,J=4.0Hz,1H),4.45(d,J=4.0Hz,1H),4.08(s,3H),4.00(s,3H),3 .84(s,3H),2.60–2.50(m,1H),2.55(s,3H),2.54–2.23(m,4H),1.85–1.75(m,2H ),1.50–1.40(m,2H),0.57(t,J=8.4Hz,2H),-0.01(s,9H).HRMS(ESI)Calcd.for C 28 H 37 NO7Si[(M+H) + ]528.2412,found 528.2419.

[0078] Example 12: Synthesis of Compound S12

[0079] Narcotine I (250.2 mg, 0.61 mmol, 1.0 equiv.) was dissolved in 15.0 mL of chloroform, and m-chloroperoxybenzoic acid (200.3 mg, 1.17 mmol, 1.9 equiv.) was added in batches at -5 °C. The reaction was stopped after 5 h. 15.0 mL of glacial chloroform and 5.0 mL of glacial isopropanol were added to the system, and the system was washed with 30.0 mL of 10% glacial sodium hydroxide solution, 30.0 mL of ice water and 30.0 mL of 10% hydrochloric acid solution in sequence. Anhydrous sulfuric acid was added. The crude product was dried over sodium sulfate, filtered, concentrated, and spun down to dryness. The resulting crude product was dissolved in 20.0 mL of methanol, and ferrous sulfate heptahydrate (400.2 mg, 1.44 mmol, 1.4 equiv.) was added at -5°C. The reaction was allowed to proceed for 12 h. The methanol was then removed by rotary evaporation. The crude product was dissolved in 15.0 mL of chloroform and washed with 0.1 mol / L ethylenediaminetetraacetic acid aqueous solution (3 × 20.0 mL) and 1 mol / L sodium hydroxide aqueous solution (2 × 20.0 mL), respectively. The product was dried over anhydrous sodium sulfate, filtered, concentrated, and spun down to dryness to obtain the crude product. Purification by silica gel column chromatography (PE:EA = 2:1) afforded 139.8 mg of XIII as a pale yellow solid in a 57% yield. XIII (400.2 mg, 1.00 mmol, 1.0 equiv.) was weighed and dissolved in 5.0 mL of dichloromethane. Triphosgene (100.1 mg, 0.34 mmol, 0.3 equiv.) and triethylamine (46.1 μL, 0.34 mmol, 0.3 equiv.) were added sequentially. The mixture was stirred for 2 h, and the reaction progress was monitored by TLC. After completion, the mixture was diluted with 5.0 mL of anhydrous dichloromethane. Dimethylamino hydrochloride (803.2 mg, 10.04 mmol, 10 equiv.) and triethylamine (1.4 mL, 10.03 mmol, 10 equiv.) were then added sequentially. The mixture was stirred for 3 h, and the reaction progress was monitored by TLC. After completion of the reaction, 30 mL of 10% HCl solution was added dropwise to the reaction system, followed by extraction and separation. The organic phase was then washed with 30.0 mL of aqueous solution and 30.0 mL of aqueous sodium carbonate solution, dried over anhydrous sodium sulfate, filtered, concentrated, and spun down to obtain a crude product, which was then purified by silica gel column chromatography (PE:EA = 1:1) to afford XIV as a white solid. Compound S1 (470.4 mg, 1.00 mmol, 1.0 equiv.) was dissolved in 6.0 mL of acetic acid, followed by the addition of N-bromosuccinimide (201.2 mg, 1.12 mmol, 1.1 equiv.). The reaction was allowed to react for 1 h, and the reaction progress was monitored by TLC. After completion of the reaction, the pH was adjusted to 10 with aqueous ammonia in an ice bath, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and spun down to obtain a crude product, which was then purified by silica gel column chromatography (PE:EA = 1:1) to afford XV as a white solid.Using compound XV and 2-(bromoethyl)trimethylsilane as reaction raw materials, according to the preparation method of Example 9, a light yellow solid S12 can be obtained with a yield of 16%. 1 H NMR (400MHz, CDCl3) δ6.97(d,J=8.3Hz,1H),6.39(d,J=8.2Hz,1H),5.90–5.86(m,2H),5.84(d,J=4.0Hz,1H),5.80(d,J=4.0Hz,1H),4.03(s,3H),3. 84(s,3H),3.82(s,3H),3.38–3.26(m,1H),2.78(s,6H),2.63–2.24(m,4H) ,2.09–1.92(m,1H),0.74–0.53(m,2H),0.02(s,9H).HRMS(ESI)Calcd.for C 29 H 38 N2O8Si[(M+H) + ]571.2470,found 571.2477.

[0080] Example 13: Synthesis of Compound S13

[0081] Using compound XV and compound V as reaction raw materials, according to the preparation method of Example 1, a white solid S13 can be prepared with a yield of 9%. 1 HNMR (400MHz, CDCl3) δ6.94 (d, J = 8.2 Hz, 1H), 6.26 (d, J = 8.2 Hz, 1H), 5.93 (d, J = 4. 2Hz,1H),5.89–5.84(m,3H),4.06(s,3H),3.97(s,3H),3.86(s,3H),3.23–3.15(m ,1H),2.79(s,6H),2.66–2.53(m,1H),2.43–2.33(m,2H),1.78–1.58(m,5H),1.25 –1.07(m,5H),0.92–0.84(m,1H),0.26(s,3H),0.22(s,3H).HRMS(ESI)Calcd.for C 32 H 42 N2O8Si[(M+H) + ]611.2783,found 611.2789.

[0082] Example 14

[0083] In this example, it was found that the synthesized silanacrotine derivative had anti-leukemia activity, which was verified by pharmacological experiments. Cell viability was analyzed using the CCK-8 method, and cell apoptosis was detected using the Annexin-V / PI double staining method. The experimental steps are as follows:

[0084] 1. Sample preparation: Dissolve S1-S13 synthesized in the above examples and Narcotine (Tianjin Chemical Industry Development Co., Ltd., Shanghai, China) (as a negative control) in DMSO (purchased from Merck) and prepare 20 mM stock solutions according to the molecular weight of the compounds.

[0085] 2. Human leukemia cell lines

[0086] KOPT-K1, MOLT-4 (T-ALL cell lines), THP1, MOLM-13 (AML cell lines), NALM-6, SEM (B-ALL cell lines);

[0087] 3. Culture medium

[0088] RPMI 1640 + 10% FBS + 1% penicillin-streptomycin mixed solution;

[0089] 4. Other materials

[0090] Full-wavelength multifunctional microplate reader: VarioskanFlash model, manufactured by Thermo Scientific, imported 96-well plates, etc.

[0091] 5. Experimental methods:

[0092] (1) CCK-8 assay for cell viability: Cells were seeded into a 96-well plate at a density of 2 x 10^5 cells / mL, with 100 μL per well. Compound concentrations were diluted in multiples starting from 20 μM, and a DMSO-only control group was also established. After incubation at 37°C, 5% CO₂ for 48 h, 10 μL of CCK-8 solution was added to each well, incubated for 2-4 h, and the OD value at 450 nm was measured using a multifunctional microplate reader. The data were processed and analyzed to determine cell viability.

[0093] (2) Annexin-V / PI double staining assay for apoptosis: Cells were seeded into 24-well plates at a density of 2*10^5 cells / mL, with 1 mL per well and a compound concentration of 2 μM. After 24 h or 48 h of treatment, the cells were collected, the culture medium was washed with PBS, and each sample was resuspended in 100 μL of 1× Binding Buffer. 2 μL of Annexin-V and PI antibodies were added, and the cells were incubated in the dark for 15 minutes. After the incubation, 300 μL of 1× Binding Buffer was added to terminate the labeling. Cell apoptosis was detected by flow cytometry and plotted as a bar graph.

[0094] Table 1 The inhibitory activity of noscapine and silanocapine on KOPT-K1 cell proliferation

[0095]

[0096] Table 1 shows the cell viability results of the T-ALL cell line KOPT-K1 treated with 20 μM Noscapine and silyl-substituted Noscapine derivatives. 1. Most silyl-substituted Noscapine derivatives exhibited superior activity to Noscapine. 2. Silyl-substituted Noscapine exhibited superior activity to directly substituted Noscapine. The activity was related to the distance between the silyl group and the benzene ring, with optimal activity achieved when the distance was three carbon atoms (S9-S11). 3. Replacing the nitrogen methyl group of Noscapine with a carbamoyl group significantly decreased activity (S12-S13). Therefore, we further validated the activity of the preferred compounds S10 (ZW004) and S11 (ZW009).

[0097] Table 2 IC values ​​of different silanacrotine derivatives in inhibiting proliferation of various leukemia cells 50

[0098]

[0099] Figure 3 , Table 2 and Figure 4 The results showed that compared with the negative control (Narcotine), S11 (ZW009) and S10 (ZW004) exhibited excellent proliferation inhibitory activity against different types of leukemia cells; at the same time, they could significantly induce apoptosis of leukemia cells, and had good development prospects.

[0100] The above embodiments are preferred implementations of the present invention, but the implementations of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be considered as equivalent replacement methods and also fall within the scope of protection of the present invention and are included in the scope of protection of the present invention.

Claims

1. A silanacrotine derivative, characterized in that: The general structural formula is as follows: R in the general structural formula Ⅰ and Ⅱ 1 and R 3 The group is selected from alkyl, deuterated alkyl; R 2 The group is selected from alkyl, deuterated alkyl, cycloalkyl, deuterated cycloalkyl, and aryl; or R in the general structural formula III and IV 4 and R 5 The group is selected from alkyl and deuterated alkane, and n is any number from 1 to 6.

2. A silanacrotine derivative according to claim 1, characterized in that: The structural formula of the specific compound is as follows:

3. A silanacrotine derivative according to claim 1, characterized in that: The preparation method of S1 is as follows: 1) Using liquid bromine as the bromination reagent, the 9′ position of Narcotine is brominated in the presence of hydrobromic acid to obtain the intermediate 9′-bromonarcotine II; 2) Under the action of methyllithium, hexamethyldisilane generates trimethylsilyllithium intermediate, which then reacts with isopropyl borate to form trimethylsilylpinacol borate III; 3) 9′-Bronacotine II and trimethylsilylpinacol borate undergo a coupling reaction in the presence of tetrakistriphenylphosphine palladium and potassium carbonate as a base to produce compound S1.

4. A silanacrotine derivative according to claim 2, characterized in that: The preparation method of S2–S8 is as follows: 1) Hydrosilane reacts with biboronic acid pinacol ester under the catalysis of palladium on carbon (Pt / C) to generate silyl pinacol borate esters IV-XII; 2) Using potassium carbonate as a base, 9′-bromonacotine II and silylpinacol borate IV-XII undergo coupling reaction under the catalysis of tetrakistriphenylphosphine palladium to produce compounds S2-S8.

5. A silanacrotine derivative according to claim 2, characterized in that: The preparation method of S9-S11 is as follows: Using nickel iodide as catalyst, 4,4′-di-tert-butylbipyridine as ligand, zinc powder as reducing agent, magnesium chloride and pyridine as additives, 9′-bromonacotine II and trimethylsilyl bromide underwent Negishi coupling to produce compounds S9-S11.

6. A silanacrotine derivative according to claim 2, characterized in that: The preparation method of S12 is as follows: 1) Narcotine I is oxidized by m-chloroperbenzoic acid (m-CPBA) and ferrous sulfate (Fe2SO4)-mediated reductive demethylation to generate intermediate XIII; 2) Intermediate XII reacts with triphosgene (BTC) under alkaline conditions, and then reacts with dimethylamine to obtain intermediate XIV; 3) XIV undergoes bromination reaction with N-bromosuccinimide in acetic acid to obtain intermediate XV; 4) Using nickel iodide as a catalyst, 4,4′-di-tert-butylbipyridine as a ligand, zinc powder as a reducing agent, magnesium chloride and pyridine as additives, XV and trimethylsilyl bromide undergo Negishi coupling to produce compound S12.

7. A silanacrotine derivative according to claim 2, characterized in that: The preparation method of S13 is as follows: Using potassium carbonate as a base, compound XV and compound IV undergo a coupling reaction catalyzed by tetrakistriphenylphosphine palladium to produce compound S13.

8. Use of the silanacrotine derivative according to any one of claims 1 to 7 in the preparation of anti-leukemia drugs.

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