Beta-carboline-thiazole compound, preparation method thereof and application of beta-carboline-thiazole compound as drug resistance reversal agent

Abstract

The invention discloses a beta-carboline-thiazole compound, a preparation method thereof and application of the beta-carboline-thiazole compound as a drug resistance reversal agent, and belongs to the technical field of medicinal chemistry. The structure of the beta-carboline-thiazole compound is shown in the specification, Boc-D-tryptophan is used as a substrate, and the beta-carboline-thiazole compound is synthesized through the steps of protecting group removal, Pictet-Spengler cascade reaction, aromatization, intermolecular polycondensation and the like. The beta-carboline-thiazole compound disclosed by the invention has the beneficial effects that the beta-carboline-thiazole compound is novel in structure, good in safety and higher in drug resistance reversal activity to human oral epidermoid cancer cell lines KBV, the raw materials of the preparation method are cheap and easy to obtain, the preparation process is simple and convenient, and the beta-carboline-thiazole compound is suitable for large-scale industrial production; and # imgabs0 #.

Description

Technical Field

[0001] The present invention relates to a class of compounds and preparation methods and applications thereof, in particular to β-carboline-thiazole compounds and preparation methods thereof and applications thereof as drug resistance reversal agents, belonging to the technical field of medicinal chemistry. Background Art

[0002] Multidrug resistance (MDR) remains a key obstacle to effective cancer chemotherapy. Increasing the efflux of multiple chemotherapeutic drugs through the transmembrane ATP-binding cassette (ABC) transporter superfamily is considered a key driver of multidrug resistance in cancer, with the role of P-glycoprotein (P-gp / ABCB1) being widely demonstrated. Combining P-gp drug efflux inhibitors with anticancer drugs that are P-gp transport substrates is considered an anticancer therapeutic strategy to overcome drug resistance by blocking P-gp-mediated drug efflux. In recent years, scientists have conducted extensive research and have screened for non-toxic, highly selective, and effective P-gp efflux inhibitors.

[0003] β-Carboline is an indole alkaloid containing a tricyclic pyridine [3,4-b] indole structure. Since the first β-carboline was isolated from Sophora flavescens in 1841, research into the isolation and synthesis of β-carbolines and their derivatives has rapidly advanced over the centuries. β-Carboline derivatives have become promising lead structures for the development of compounds with anticancer activity due to their abundant natural sources, high structural diversity, rapid chemical reactivity, and ability to interact with a variety of anticancer targets, such as DNA intercalation and groove binding, enzymes such as GPX4, topoisomerases, and kinases, and proteins such as tubulin and ABCG2 / BCRP.

[0004] Thiazoles have a unique chemical structure and can be modified with various functional groups to form a wide range of derivatives. These derivatives exhibit significant activity in interacting with transporters such as P-gp, effectively inhibiting drug efflux and thus reversing drug resistance.

[0005] Although β-carboline compounds themselves have received widespread attention, there are relatively few studies on β-carboline-thiazole skeleton compounds, especially for β-carboline skeleton compounds with a thiazole substituent at the C-3 position. No relevant research reports have been seen so far, and their drug resistance activity is still unknown. Summary of the Invention

[0006] The purpose of the present invention is to innovatively integrate the two pharmacophores of β-carboline and thiazole to provide a new type of β-carboline-thiazole compound with a new skeleton, as well as a preparation method and drug-resistant application of the compound.

[0007] In order to achieve the above objectives, the present invention adopts the following technical solutions: β-Carboline-thiazole compounds, the structure is shown below:

[0008] Wherein, R1 is any one of the following structures: 、 、 、 、 、 ; R2 is any one of the following structures: 、 .

[0009] The preparation method of the aforementioned β-carboline-thiazole compound, wherein R2 of the β-carboline-thiazole compound is The preparation method of the β-carboline-thiazole compound comprises the following steps: Step 1: Dissolve Boc-D-tryptophan in dichloromethane, add pentafluorophenyl diphenylphosphinate and triethylamine in sequence, stir at room temperature, add dimethyl azidocystine and triphenylphosphine, react at 45°C for 5h, remove the solvent by vacuum rotary evaporation to obtain a thiazoline product, dissolve the obtained thiazoline product in dichloromethane, add 1,8-diazabicyclo[5,4,0]undec-7-ene and trichlorobromomethane, react at room temperature for 2h, after completion of the reaction, quench the reaction, extract with dichloromethane and combine the organic phases, dry the organic phases, remove the solvent, and use V 石油醚 :V 乙酸乙酯 =5:1, and the solvent was removed to obtain compound 3 as a yellow oil; Step 2: Dissolve compound 3 in dichloromethane, add benzaldehyde compound or glyoxal dimethyl acetal at room temperature, the benzaldehyde compound is any one of p-methoxybenzaldehyde, trimethoxybenzaldehyde, p-dimethylaminobenzaldehyde, 3-trifluoromethylbenzaldehyde and 4-cyanobenzaldehyde, add trifluoroacetic acid dropwise at 0°C, react at room temperature for 2h, after the reaction is completed, quench the reaction, extract with dichloromethane and combine the organic phases, dry the organic phases, remove the solvent, dissolve the product in tetrahydrofuran, add potassium permanganate, react at room temperature for 3h, and use V 石油醚 :V 乙酸乙酯 =3:1 or 2:1, pass through a column, and dry.

[0010] The preparation method of the aforementioned β-carboline-thiazole compound, wherein R2 of the β-carboline-thiazole compound is The preparation method of the β-carboline-thiazole compound comprises the following steps: Step 1: Dissolve Boc-D-tryptophan in dichloromethane, add pentafluorophenyl diphenylphosphinate and triethylamine in sequence, stir at room temperature, add dimethyl azidocystine and triphenylphosphine, react at 45°C for 5h, remove the solvent by vacuum rotary evaporation to obtain a thiazoline product, dissolve the obtained thiazoline product in dichloromethane, add 1,8-diazabicyclo[5,4,0]undec-7-ene and trichlorobromomethane, react at room temperature for 2h, after completion of the reaction, quench the reaction, extract with dichloromethane and combine the organic phases, dry the organic phases, remove the solvent, and use V 石油醚 :V 乙酸乙酯 =5:1, and the solvent was removed to obtain compound 3 as a yellow oil; Step 2: Dissolve compound 3 in dichloromethane, add benzaldehyde compound or glyoxal dimethyl acetal at room temperature, the benzaldehyde compound is any one of p-methoxybenzaldehyde, trimethoxybenzaldehyde, p-dimethylaminobenzaldehyde, 3-trifluoromethylbenzaldehyde and 4-cyanobenzaldehyde, add trifluoroacetic acid dropwise at 0°C, react at room temperature for 2h, after the reaction is completed, quench the reaction, extract with dichloromethane and combine the organic phases, dry the organic phases, remove the solvent, dissolve the product in tetrahydrofuran, add potassium permanganate, react at room temperature for 3h, and use V 石油醚 :V 乙酸乙酯 =3:1 or 2:1, pass through column, dry and obtain intermediate product A; Step 3: Dissolve the intermediate product A in a mixed solution of tetrahydrofuran and methanol, then add a lithium hydroxide aqueous solution, react at room temperature for 10 hours, adjust the pH to 3.5, extract with ethyl acetate and water, back-extract the aqueous phase with ethyl acetate, combine the organic phases and spin-dry to obtain the intermediate product B; Step 4: Dissolve the intermediate product B in dichloromethane, then add 2-(7-azabenzotriazole)-N,N,N',N',-tetramethyluronium hexafluorophosphate, 1-hydroxybenzotriazole and N,N-diisopropylethylamine, react at room temperature for 10 minutes, then add 5-chlorotryptamine hydrochloride, react at room temperature for 15 hours, extract with dilute hydrochloric acid and dichloromethane, back-extract the aqueous phase with dichloromethane, combine the organic phases and extract with sodium hydroxide solution, back-extract the aqueous phase with dichloromethane, combine the organic phases and spin-dry, and use V 石油醚 :V 乙酸乙酯 =4:1 pass through column and dry; .

[0011] Preferably, in step 1, the molar ratio of Boc-D-tryptophan, pentafluorophenyldiphenylphosphinate, triethylamine, dimethyl azidocystine, triphenylphosphine, 1,8-diazabicyclo[5,4,0]undec-7-ene and bromotrichloromethane is 4:4:8:1:10:6:6.

[0012] Preferably, in step 2, the molar ratio of compound 3 to dialdehyde dimethyl acetal, p-anisaldehyde, trimethoxybenzaldehyde, p-dimethylaminobenzaldehyde, 3-trifluoromethylbenzaldehyde or 4-cyanobenzaldehyde is 2:3.

[0013] Preferably, in step 3, the molar ratio of intermediate product A to lithium hydroxide is 2:15.

[0014] Preferably, in step 4, the molar ratio of the intermediate product B, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, 1-hydroxybenzotriazole, N,N-diisopropylethylamine and 5-chlorotryptamine hydrochloride is 20:26:1:72:30.

[0015] The use of the aforementioned β-carboline-thiazole compound in the preparation of a drug resistance reversal agent for human oral epidermoid carcinoma cells, wherein the β-carboline-thiazole compound is any one of the following structures:

[0016] The present invention is beneficial in that: (1) The β-carboline-thiazole compounds provided by the present invention have novel structures, good safety, and higher drug resistance reversal activity against human oral epidermoid carcinoma cell line KBV than verapamil; (2) The preparation method of the β-carboline-thiazole compound provided by the present invention has cheap and readily available raw materials, a simple preparation process, and is suitable for large-scale industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 The IC value of paclitaxel alone against drug-resistant human oral cancer cell line KBV 50 Value evaluation chart; Figure 2 The IC value of paclitaxel against drug-resistant human oral cancer cell line KBV under the condition of co-administration of verapamil and paclitaxel is 50 Value evaluation chart; Figure 3 The IC value of paclitaxel against drug-resistant human oral cancer cell line KBV under the condition of co-administration of compound 4 and paclitaxel is 50 Value evaluation chart; Figure 4 The IC value of paclitaxel against drug-resistant human oral cancer cell line KBV under the condition of co-administration of compound 5 and paclitaxel is 50 Value evaluation chart; Figure 5The IC value of paclitaxel against drug-resistant human oral cancer cell line KBV under the condition of co-administration of compound 9 and paclitaxel is 50 Value evaluation chart; Figure 6 The IC value of paclitaxel against drug-resistant human oral cancer cell line KBV under the condition of co-administration of compound 11 and paclitaxel is 50 Value evaluation chart. DETAILED DESCRIPTION

[0018] The present invention is described in detail below with reference to specific embodiments.

[0019] 1. Structure of β-carboline-thiazole compounds The present invention innovatively combines the two pharmacophores of β-carboline and thiazole, and provides a β-carboline-thiazole compound with the following structure:

[0020] Wherein, R1 is any one of the following structures:

[0021] 、 、 、 、 、 ; R2 is any one of the following structures: 、 .

[0022] 2. Preparation Method of β-Carboline-Thiazole Compounds The preparation method of the above-mentioned β-carboline-thiazole compound provided by the present invention is: using Boc-D-tryptophan as a substrate, and completing the synthesis of the above-mentioned β-carboline-thiazole compound through the steps of removing the protecting group, Pictet-Spengler cascade reaction, aromatization, intermolecular polycondensation, etc.

[0023] Example 1

[0024] Step 1: Dissolve commercially available Boc-D-tryptophan (compound 1, 1 mmol) in dry dichloromethane (DCM, 50 mL), add pentafluorophenyl diphenylphosphinate (FDPP, 1 mmol) and triethylamine (Et3N, 2 mmol) in sequence, stir at room temperature for 5 min, add dimethyl azidocystine (compound 2, 0.25 mmol) and triphenylphosphine (PPh3, 2.5 mmol), react at 45 ° C for 5 h, and remove the solvent by vacuum rotary evaporation to obtain the thiazoline product without purification. The obtained thiazoline product was dissolved in dry DCM (50 mL), and 1,8-diazabicyclo[5,4,0]undec-7-ene (DBU, 1.5 mmol) and bromotrichloromethane (CBrCl3, 1.5 mmol) were added. The reaction was allowed to proceed at room temperature for 2 h. After completion of the reaction, saturated ammonium chloride solution (80 mL) was added to the reaction system to quench the reaction. The mixture was extracted with dichloromethane (80 mL each time, 3 times) and the organic phases were combined. The organic phase was dried over anhydrous sodium sulfate, and the solvent was removed by vacuum rotary evaporation. The mixture was then dried with V 石油醚 :V 乙酸乙酯 =5:1, and the solvent was removed by column evaporation under vacuum to obtain a yellow oil (compound 3) with a yield of 79%.

[0025] Compound 3 1 The data of H NMR and ESI-HRMS are as follows: 1 H NMR (400MHz, CDCl3) δ 8.14 (s, 1H), 8.03 (s, 1H), 7.45 (d, J= 7.6 Hz, 1H), 7.35 (d, J= 7.6 Hz, 1H), 7.18 (dd, J= 7.6Hz, J= 7.6 Hz, 1H), 7.08 (dd, J= 7.6Hz, J= 7.6 Hz, 1H), 6.69 (d, J= 0.8Hz, 1H), 5.39 (s, br, 2H), 3.98 (s, 3H), 3.55 (d, J= 13.2 Hz, 1H), 3.45 (dd, J= 4.0Hz, J= 13.2Hz, 1H), 1.28 (s, 9H); ESI-HRMS: C 20 H 23 N3O4S([M+Na] + ) Calculated value 424.1302, theoretical value 423.2308.

[0026] Step 2: Compound 3 (1 mmol) was dissolved in dry DCM (20 mL), p-anisaldehyde (1.5 mmol) was added at room temperature, trifluoroacetic acid (TFA, 1 mL) was added dropwise at 0 ° C, and the reaction was carried out at room temperature for 2 h. After the reaction was completed by TLC monitoring, saturated sodium bicarbonate solution (50 mL) was added to the reaction system to quench the reaction, and the mixture was extracted with dichloromethane (40 mL each time, 3 times) and the organic phases were combined. The organic phases were dried over anhydrous sodium sulfate, and the solvent was removed by vacuum rotary evaporation. The product was dissolved in dry tetrahydrofuran (THF, 30 mL), potassium permanganate (KMnO4, 1.5 mmol) was added, and the reaction was carried out at room temperature for 3 h. 石油醚 :V 乙酸乙酯 =3:1, passed through a column, and dried to obtain a light yellow solid (Compound 4) with a yield of 75%.

[0027] Compound 4 1 H NMR, 13 The C NMR and ESI-HRMS data are as follows: 1 H NMR (500MHz, CDCl3) δ 8.75 (s, 1H), 8.11 (s, 1H), 8.01 (d, J =8Hz,1H),7.87(d, J =8Hz, 2H), 7.50(d, J =8.5Hz,1H),7.45(t, J =8Hz,1H),7.22(t, J =7.5Hz, 1H), 6.96(d, J =8.5Hz, 2H), 3.92 (s, 3H), 3.76 (s, 3H); 13 C NMR (125MHz, CDCl3) δ 162.4, 160.7, 147.5, 141.9, 141.3, 134.1, 130.8, 129.8, 129.2, 129.0, 122.3, 121.1, 114.8, 112.3, 110.7, 55.5, 52.6; ESI-HRMS: C 23 H 17 N3NaO3S([M+Na] + ) Calculated value 438.0883, theoretical value 438.0880.

[0028] Example 2

[0029] Step 1: It is exactly the same as step 1 of Example 1 and will not be repeated here.

[0030] Step 2: Compound 3 (1 mmol) was dissolved in dry DCM (20 mL), trimethoxybenzaldehyde (1.5 mmol) was added at room temperature, TFA (1 mL) was added dropwise at 0 ° C, and the reaction was carried out at room temperature for 2 h. After the reaction was completed by TLC monitoring, saturated sodium bicarbonate solution (50 mL) was added to the reaction system to quench the reaction, and the mixture was extracted with dichloromethane (40 mL each time, 3 times) and the organic phases were combined. The organic phase was dried over anhydrous sodium sulfate, and the solvent was removed by vacuum rotary evaporation. The product was dissolved in dry THF (30 mL), KMnO4 (1.5 mmol) was added, and the reaction was carried out at room temperature for 3 h. 石油醚 :V 乙酸乙酯 =3:1, passed through a column, and dried to obtain a light yellow solid (Compound 5) with a yield of 67%.

[0031] Compound 5 1 H NMR, 13 The C NMR and ESI-HRMS data are as follows: 1 H NMR (500 MHz, CDCl3) δ 9.20 (s, 1H), 8.85 (s, 1H), 8.19 (s, 1H), 8.09 (d, J =8.0Hz, 1H), 7.57(d, J =5.0Hz, 2H), 7.34-7.31 (m, 1H), 7.18 (s, 2H), 4.00 (s, 3H), 3.90 (s, 3H), 3.89 (s, 6H); 13 C NMR (126MHz, CDCl3) δ 172.0, 162.5, 153.9, 147.4, 142.4, 141.2, 140.9, 139.0, 134.5, 133.5, 130.8, 1 29.1, 128.9, 122.4, 122.4, 121.1, 112.1, 111.0, 105.8, 61.1, 56.5, 52.6, 29.8; ESI-HRMS: C 25 H 21 N3NaO5S([M+Na] + ) Calculated value 498.1094, theoretical value 498.1089.

[0032] Example 3

[0033] Step 1: It is exactly the same as step 1 of Example 1 and will not be repeated here.

[0034] Step 2: Compound 3 (1 mmol) was dissolved in dry DCM (20 mL), p-dimethylaminobenzaldehyde (1.5 mmol) was added at room temperature, TFA (1 mL) was added dropwise at 0 ° C, and the reaction was carried out at room temperature for 2 h. After the reaction was completed by TLC monitoring, saturated sodium bicarbonate solution (50 mL) was added to the reaction system to quench the reaction, and the mixture was extracted with dichloromethane (40 mL each time, 3 times) and the organic phases were combined. The organic phases were dried over anhydrous sodium sulfate, and the solvent was removed by vacuum rotary evaporation. The product was dissolved in dry THF (30 mL), KMnO4 (1.5 mmol) was added, and the reaction was carried out at room temperature for 3 h. 石油醚 :V 乙酸乙酯 =2:1, passed through a column, and dried to obtain a light yellow solid (Compound 6) with a yield of 65%.

[0035] Compound 6 1 H NMR, 13 The C NMR and ESI-HRMS data are as follows: 1 H NMR (400MHz, CDCl3) δ 8.82 (d, J =10.0Hz, 2H), 8.18 (s, 1H), 8.07 (d, J =7.6Hz,1H),7.90(d, J =8.8Hz, 2H), 7.56-7.47 (m, 2H), 7.28 (m, 1H), 6.83 (d, J =8.8Hz, 2H), 3.98 (s, 3H), 3.00 (s, 6H); 13 C NMR (100MHz, CDCl3) δ 172.6, 162.5, 151.0, 147.4, 142.9, 140.9, 140.8, 134.1, 130.3, 129.2 , 128.9, 128.6, 122.5, 122.2, 120.8, 112.6, 111.9, 109.8, 52.5, 40.4; ESI-HRMS: C 24 H 20 N4NaO2S([M+Na] + ) Calculated value 451.1199, theoretical value 451.1200.

[0036] Example 4

[0037] Step 1: It is exactly the same as step 1 of Example 1 and will not be repeated here.

[0038] Step 2: Compound 3 (1 mmol) was dissolved in dry DCM (20 mL), glyoxal dimethyl acetal (1.5 mmol) was added at room temperature, TFA (1 mL) was added dropwise at 0 ° C, and the reaction was carried out at room temperature for 2 h. After the reaction was completed by TLC monitoring, saturated sodium bicarbonate solution (50 mL) was added to the reaction system to quench the reaction, and the mixture was extracted with dichloromethane (40 mL each time, 3 times) and the organic phases were combined. The organic phase was dried over anhydrous sodium sulfate, and the solvent was removed by vacuum rotary evaporation. The product was dissolved in dry THF (30 mL), KMnO4 (1.5 mmol) was added, and the reaction was carried out at room temperature for 3 h. 石油醚 :V 乙酸乙酯 =2:1, passed through a column, and dried to obtain a light yellow solid (Compound 7) with a yield of 67%.

[0039] Compound 7 1 H NMR, 13 The C NMR and ESI-HRMS data are as follows: 1 H NMR (500MHz, CDCl3) δ 9.32 (s, 1H), 8.97 (s, 1H), 8.21 (s, 1H), 8.15 (d, J =8.0Hz, 1H), 7.57-7.52 (m, 2H), 7.29 (t, J =6.5Hz, 1H), 5.69 (s, 1H), 4.00 (s, 3H), 3.56 (s, 6H); 13 C NMR (125MHz, CDCl3) δ 171.6, 162.3, 147.4, 140.8, 134.0, 139.9, 134.3, 130.9, 129.1, 128.8, 122.1, 121.5, 120.6, 112.1, 111.8, 106.7, 54.8, 53.5, 52.5, 29.7; ESI-HRMS: C 19 H 17 N3NaO4S([M+Na] + ) Calculated value 406.0832, theoretical value 406.0830.

[0040] Example 5

[0041] Step 1: It is exactly the same as step 1 of Example 1 and will not be repeated here.

[0042] Step 2: Compound 3 (1 mmol) was dissolved in dry DCM (20 mL), 3-trifluoromethylbenzaldehyde (1.5 mmol) was added at room temperature, TFA (1 mL) was added dropwise at 0 ° C, and the reaction was carried out at room temperature for 2 h. After the reaction was completed by TLC monitoring, saturated sodium bicarbonate solution (50 mL) was added to the reaction system to quench the reaction, and the mixture was extracted with dichloromethane (40 mL each time, 3 times) and the organic phases were combined. The organic phase was dried over anhydrous sodium sulfate, and the solvent was removed by vacuum rotary evaporation. The product was dissolved in dry THF (30 mL), KMnO4 (1.5 mmol) was added, and the reaction was carried out at room temperature for 3 h. 石油醚 :V 乙酸乙酯 =2:1, passed through a column, and dried to obtain a light yellow solid (Compound 8) with a yield of 74%.

[0043] Compound 8 1 H NMR, 13 The C NMR and ESI-HRMS data are as follows: 1 H NMR (500 MHz, d -DMSO)δ 11.96 (s, 1H), 8.87 (s, 1H), 8.48 (s, 1H), 8.44 (d, J =8Hz,1H),8.37(d, J =7.5Hz, 2H), 7.93(d, J =8Hz,1H),7.88(t, J =8Hz, 1H), 7.68(d, J =8Hz, 1H), 7.61(t, J =8Hz, 1H), 7.32(t, J =7.5Hz, 1H), 2.53 (s, 3H); 13 C NMR (125 MHz, d -DMSO)δ 170.93, 161.81, 147.13, 142.32, 140.23, 140.17, 138.59, 134.37, 132.81, 131.11, 130.50, 130 .06, 129.41, 126.03, 125.77, 125.47, 123.60, 122.73, 121.58, 120.79, 113.06, 111.06, 52.43; ESI-HRMS: C 23 H 14 F3N3NaO2S([M+Na] +) Calculated value 476.0651, theoretical value 476.0649.

[0044] Example 6

[0045] Step 1: It is exactly the same as step 1 of Example 1 and will not be repeated here.

[0046] Step 2: Compound 3 (1 mmol) was dissolved in dry DCM (20 mL), 4-cyanobenzaldehyde (1.5 mmol) was added at room temperature, TFA (1 mL) was added dropwise at 0 ° C, and the reaction was carried out at room temperature for 2 h. After the reaction was completed by TLC monitoring, saturated sodium bicarbonate solution (50 mL) was added to the reaction system to quench the reaction, and the mixture was extracted with dichloromethane (40 mL each time, 3 times) and the organic phases were combined. The organic phases were dried over anhydrous sodium sulfate, and the solvent was removed by vacuum rotary evaporation. The product was dissolved in dry THF (30 mL), KMnO4 (1.5 mmol) was added, and the reaction was carried out at room temperature for 3 h. 石油醚 :V 乙酸乙酯 =2:1, passed through a column, and dried to obtain a light yellow solid (Compound 9) with a yield of 75%.

[0047] Compound 9 1 H NMR, 13 The C NMR and ESI-HRMS data are as follows: 1 H NMR (500 MHz, d- DMSO) δ 11.92 (s, 1H), 8.88 (s, 1H), 8.49 (s, 1H), 8.44 (d, J =8.0Hz,1H),8.22(d, J =8.0Hz, 2H), 8.08(d, J =8.5Hz, 2H), 7.64(d, J =8.0Hz,1H),7.59(t, J =8.0Hz,1H),7.29(t, J =7.5Hz, 1H), 3.89 (s, 3H); 13 C NMR (125 MHz, d- DMSO)δ 170.8, 161.8, 147.2, 142.3, 141.9, 140.2, 139.8, 134.5, 133.2, 131.3, 130.6, 129.7, 129.5, 122.8, 121.5, 120.9, 119.3, 113.1, 111.8, 111.4, 79.6, 52.5; ESI-HRMS: C 23 H 14 N4NaO2S([M+Na] + ) Calculated value 433.0728, theoretical value 433.0730.

[0048] Example 7

[0049] Step 1: Dissolve the compound 4 (1 mmol) prepared in Example 1 in a mixed solution of THF and methanol (MeOH) (V THF :V MeOH = 1:1), and then added lithium hydroxide (LiOH) aqueous solution (concentration 0.5 mmol / mL, amount 15 mL), and reacted at room temperature for 10 h. After TLC detection, the pH was adjusted to 3.5 with dilute hydrochloric acid (concentration 1 M), and then extracted with ethyl acetate and water. The aqueous phase was back-extracted twice with ethyl acetate, and the organic phases were combined and dried to obtain compound 10, which was directly used in the next reaction without purification.

[0050] Step 2: Compound 10 (1 mmol) was dissolved in dry DCM (50 mL), and then 2-(7-azabenzotriazole)-N, N, N', N'-tetramethyluronium hexafluorophosphate (HATU, 1.3 mmol), 1-hydroxybenzotriazole (HOBt, 0.05 mmol) and N, N-diisopropylethylamine (DIPEA, 3.6 mmol) were added. After the reaction at room temperature for 10 min, 5-chlorotryptamine hydrochloride (1.5 mmol) was added and the reaction was continued at room temperature for 15 h. After the reaction was completed by TLC monitoring, the mixture was extracted with dilute hydrochloric acid (concentration 0.3 M) and dichloromethane, and the aqueous phase was back-extracted twice with dichloromethane. The organic phases were combined and extracted with sodium hydroxide solution (concentration 0.5 M). The aqueous phase was back-extracted twice with dichloromethane. The organic phases were combined, dried by rotation, and washed with V 石油醚 :V 乙酸乙酯 =4:1, passed through a column, and dried to obtain a white solid (Compound 11) with a yield of 83%.

[0051] Compound 11 1 H NMR, 13 The C NMR and ESI-HRMS data are as follows: 1 H NMR (500MHz, CDCl3) δ 8.71 (s, 1H), 8.63 (s, 1H), 8.22 (s, 1H), 8.21-8.19 (m, 1H), 8.13 (s, 1H), 8.03-7.97 (d, 1H), 7.73 (t, J=6.2Hz,1H),7.67(d, J =2.0Hz, 1H), 7.63-7.53 (m, 2H), 7.37 (t, J =7.3Hz, 1H), 7.32(d, J =8.6Hz, 1H), 7.15 (ddd, J =8.5, 4.7, 2.2Hz, 4H), 3.92 (s, 3H), 3.84 (q, J =6.8Hz, 3H), 3.13(t, J =7.0Hz,2H); 13 C NMR (125MHz, CDCl3) δ 161.51, 160.44, 150.58, 140.71, 134.59, 133.86, 130.41, 129.44, 128.81, 128.57, 125.22, 124.08, 123.4 5, 122.36, 122.18, 121.96, 120.71, 118.43, 114.63, 113.13, 112.05, 111.78, 109.60, 55.39, 39.72, 25.46; ESI-HRMS: C 32 H 24 ClN5O2S([M+Na] + ) Calculated value 577.1335, theoretical value 577.1339.

[0052] 3. Pharmacological studies of β-carboline-thiazole compounds The drug resistance reversal activity of compounds 4, 5, 6, 7, 8, 9 and 11 against the drug-resistant human oral cancer cell line KBV was detected.

[0053] The MTT method was used to determine the effects of the above compounds on the survival rate of drug-resistant human oral epidermoid carcinoma cell line KBV when administered alone.

[0054] The MTT assay was used to determine the effects of the above compounds on the survival rate of drug-resistant human oral epidermoid carcinoma cell line KBV and the half inhibitory concentration (IC50) when they were co-administered with paclitaxel (PTX). 50 ).

[0055] Detection principle: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) can penetrate the cell membrane and enter the cell. The succinate dehydrogenase in the mitochondria of living cells can reduce exogenous MTT to blue-purple needle-shaped formazan crystals that are insoluble in water and deposited in the cells. The crystals can be dissolved by dimethyl sulfoxide (DMSO). The absorbance of the crystals can reflect the cell survival rate. The half inhibitory concentration (IC 50 ) is defined as the drug concentration when 50% of the tumor cells survive. According to the measured optical density (OD value), a standard curve of cell survival rate is prepared, and the corresponding drug concentration is then obtained on the curve.

[0056] The determination method is as follows: (1) The drug-resistant human oral epidermoid carcinoma cell line KBV was inoculated into a 96-well plate at a density of 3000 cells / well and cultured in a CO2 incubator for 24 h; (2) PTX (stock concentration 10 mM) was serially diluted to 1000 nM, 300 nM, 100 nM, 30 nM, 10 nM, 3 nM, 1 nM, 0.3 nM, and 0.1 nM. The concentrations of compounds 4, 5, 6, 7, 8, 9, and 11 (stock concentration 10 mM) were diluted to 10 μM, and the concentration of verapamil (control compound, stock concentration 10 mM) was diluted to 10 μM. (3) Three sets of parallel wells were set for each concentration of PTX, and different concentrations of PTX were added to the seeded cells (for calculating the IC 50 ), or add different concentrations of PTX and 10 μM of the test compound together and in equal amounts to the seeded cells (for calculating the cell survival rate and the IC of PTX in the case of co-administration) 50 ), or add 10 μM of the test compound to the seeded cells (for calculating the cell survival rate in the case of single drug administration), and continue to culture for 72 h; (4) Remove the 96-well plate from the CO2 incubator, add 30 μL of MTT solution (concentration 5 mg / mL) to each well, and continue incubating in the CO2 incubator for 4 h; (5) Discard the upper liquid, add 150 μL DMSO to each well, and shake gently for 20 minutes to completely dissolve the formazan crystals; (6) Use an enzyme-labeled analyzer to measure the absorbance of each well at 490 nm and calculate the cell survival rate.

[0057] Three parallel experiments were performed and the experimental results were averaged.

[0058] The survival rates of the drug-resistant human oral epidermoid carcinoma cell line KBV measured in the case of single drug administration and co-administration are shown in Table 1.

[0059] Table 1 Survival rate of drug-resistant human oral epidermoid carcinoma cell line KBV

[0060] From Table 1 we can see that: (1) When administered alone, except for compounds 7 and 9 (the survival rate of drug-resistant human oral epidermoid carcinoma cell line KBV was less than 80%), all other compounds showed good safety. (2) Compounds 4, 5, 9, and 11, when co-administered with PTX, all showed significant inhibitory effects on the drug-resistant human oral epidermoid carcinoma cell line KBV, with cell survival rates all below 37%.

[0061] Compounds 4, 5, 9 and 11 with excellent co-administration effects were selected, and verapamil was used as a control to investigate the half-maximal inhibitory concentration (IC50) of PTX under the conditions of co-administration of these compounds with PTX. 50 )evaluate.

[0062] IC of PTX against drug-resistant human oral cancer cell line KBV under the condition of PTX alone 50 Value evaluation chart see Figure 1 .

[0063] IC of PTX against drug-resistant human oral cancer cell line KBV under the condition of co-administration of verapamil and PTX 50 Value evaluation chart see Figure 2 .

[0064] When compound 4 and PTX were co-administered, the IC of PTX against drug-resistant human oral cancer cell line KBV was 50 Value evaluation chart see Figure 3 .

[0065] When compound 5 and PTX were co-administered, the IC of PTX against drug-resistant human oral cancer cell line KBV was 50 Value evaluation chart see Figure 4 .

[0066] When compound 9 and PTX were co-administered, the IC of PTX against drug-resistant human oral cancer cell line KBV was 50 Value evaluation chart see Figure 5 .

[0067] IC of PTX against drug-resistant human oral cancer cell line KBV under the condition of co-administration of compound 11 and PTX 50 Value evaluation chart see Figure 6 .

[0068] Depend on Figure 1 The IC values ​​of PTX against drug-resistant human oral cancer cell line KBV were obtained under the condition of single administration of PTX.50 Value. Figures 2 to 6 The IC values ​​of PTX on drug-resistant human oral cancer cell line KBV were obtained under the condition of co-administration of each test compound and PTX. 50 value.

[0069] IC of PTX against drug-resistant human oral cancer cell line KBV 50 The calculation results are shown in Table 2.

[0070] Table 2 IC of PTX under co-application conditions 50 value

[0071] From Table 2 we can see that: (1) IC of PTX against drug-resistant human oral epidermoid carcinoma cell line KBV under the condition of PTX alone 50 The value is as high as 1612.0nM; (2) After PTX was co-administered with verapamil, compound 4, compound 5, compound 9, and compound 11, the IC values ​​of PTX on the drug-resistant human oral epidermoid carcinoma cell line KBV were 50 The values ​​were significantly reduced. Compared with the commercially available verapamil, compounds 4, 5, 9 and 11 all showed significant resistance-reversal activity against paclitaxel, especially compounds 5 and 9. The resistance-reversal activity of the two was more than 3 times higher than that of the commercially available drug verapamil, and 89.5 times higher than that of paclitaxel alone.

[0072] Considering that compound 9 still has a certain effect on the survival rate of the drug-resistant human oral epidermoid carcinoma cell line KBV under high concentration conditions of single administration and has cytotoxicity, therefore, compared with compound 9, compound 5 has better application prospects.

[0073] It should be noted that the above embodiments are merely examples for the purpose of clearly illustrating the present invention and are not intended to limit the embodiments of the present invention. A person skilled in the art would be able to make other variations or modifications based on the above description. It is not possible to enumerate all embodiments here. Any obvious variations or modifications arising from the technical solution of the present invention remain within the scope of protection of the present invention.

Claims

1. β-Carboline-thiazole compounds, characterized in that The structure is as follows: ； Wherein, R1 is any one of the following structures: 、 、 、 、 、 ； R2 is any one of the following structures: 、 。 2. The method for preparing the β-carboline-thiazole compound according to claim 1, wherein R2 of the β-carboline-thiazole compound is , characterized in that, The preparation method of the β-carboline-thiazole compound comprises the following steps: Step 1: Dissolve Boc-D-tryptophan in dichloromethane, add pentafluorophenyl diphenylphosphinate and triethylamine in sequence, stir at room temperature, add dimethyl azidocystine and triphenylphosphine, react at 45°C for 5h, remove the solvent by vacuum rotary evaporation to obtain a thiazoline product, dissolve the obtained thiazoline product in dichloromethane, add 1,8-diazabicyclo[5,4,0]undec-7-ene and trichlorobromomethane, react at room temperature for 2h, after completion of the reaction, quench the reaction, extract with dichloromethane and combine the organic phases, dry the organic phases, remove the solvent, and use V 石油醚 :V 乙酸乙酯 =5:1, and the solvent was removed to obtain compound 3 as a yellow oil; Step 2: Dissolve compound 3 in dichloromethane, add benzaldehyde compound or glyoxal dimethyl acetal at room temperature, the benzaldehyde compound is any one of p-methoxybenzaldehyde, trimethoxybenzaldehyde, p-dimethylaminobenzaldehyde, 3-trifluoromethylbenzaldehyde and 4-cyanobenzaldehyde, add trifluoroacetic acid dropwise at 0°C, react at room temperature for 2h, after the reaction is completed, quench the reaction, extract with dichloromethane and combine the organic phases, dry the organic phases, remove the solvent, dissolve the product in tetrahydrofuran, add potassium permanganate, react at room temperature for 3h, and use V 石油醚 :V 乙酸乙酯 =3:1 or 2:1, pass through a column, and dry.

3. The method for preparing the β-carboline-thiazole compound according to claim 1, wherein R2 of the β-carboline-thiazole compound is , characterized in that, The preparation method of the β-carboline-thiazole compound comprises the following steps: Step 1: Dissolve Boc-D-tryptophan in dichloromethane, add pentafluorophenyl diphenylphosphinate and triethylamine in sequence, stir at room temperature, add dimethyl azidocystine and triphenylphosphine, react at 45°C for 5h, remove the solvent by vacuum rotary evaporation to obtain a thiazoline product, dissolve the obtained thiazoline product in dichloromethane, add 1,8-diazabicyclo[5,4,0]undec-7-ene and trichlorobromomethane, react at room temperature for 2h, after completion of the reaction, quench the reaction, extract with dichloromethane and combine the organic phases, dry the organic phases, remove the solvent, and use V 石油醚 :V 乙酸乙酯 =5:1, and the solvent was removed to obtain compound 3 as a yellow oil; Step 2: Dissolve compound 3 in dichloromethane, add benzaldehyde compound or glyoxal dimethyl acetal at room temperature, the benzaldehyde compound is any one of p-methoxybenzaldehyde, trimethoxybenzaldehyde, p-dimethylaminobenzaldehyde, 3-trifluoromethylbenzaldehyde and 4-cyanobenzaldehyde, add trifluoroacetic acid dropwise at 0°C, react at room temperature for 2h, after the reaction is completed, quench the reaction, extract with dichloromethane and combine the organic phases, dry the organic phases, remove the solvent, dissolve the product in tetrahydrofuran, add potassium permanganate, react at room temperature for 3h, and use V 石油醚 :V 乙酸乙酯 =3:1 or 2:1, pass through column, dry and obtain intermediate product A; Step 3: Dissolve the intermediate product A in a mixed solution of tetrahydrofuran and methanol, then add a lithium hydroxide aqueous solution, react at room temperature for 10 hours, adjust the pH to 3.5, extract with ethyl acetate and water, back-extract the aqueous phase with ethyl acetate, combine the organic phases and spin-dry to obtain the intermediate product B; Step 4: Dissolve the intermediate product B in dichloromethane, then add 2-(7-azabenzotriazole)-N,N,N',N',-tetramethyluronium hexafluorophosphate, 1-hydroxybenzotriazole and N,N-diisopropylethylamine, react at room temperature for 10 minutes, then add 5-chlorotryptamine hydrochloride, react at room temperature for 15 hours, extract with dilute hydrochloric acid and dichloromethane, back-extract the aqueous phase with dichloromethane, combine the organic phases and extract with sodium hydroxide solution, back-extract the aqueous phase with dichloromethane, combine the organic phases and spin-dry, and use V 石油醚 :V 乙酸乙酯 =4:1 pass through column and dry; 。 4. The preparation method according to claim 2 or 3, characterized in that In step 1, the molar ratio of Boc-D-tryptophan, pentafluorophenyldiphenylphosphinate, triethylamine, dimethyl azidocystine, triphenylphosphine, 1,8-diazabicyclo[5,4,0]undec-7-ene and bromotrichloromethane is 4:4:8:1:10:6:

6.

5. The preparation method according to claim 2 or 3, characterized in that In step 2, the molar ratio of compound 3 to dialdehyde dimethyl acetal, p-anisaldehyde, trimethoxybenzaldehyde, p-dimethylaminobenzaldehyde, 3-trifluoromethylbenzaldehyde or 4-cyanobenzaldehyde is 2:

3.

6. The preparation method according to claim 3, characterized in that In step 3, the molar ratio of the intermediate product A to lithium hydroxide is 2:

15.

7. The preparation method according to claim 3, characterized in that In step 4, the molar ratio of intermediate B, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, 1-hydroxybenzotriazole, N,N-diisopropylethylamine and 5-chlorotryptamine hydrochloride is 20:26:1:72:

30.

8. Use of the β-carboline-thiazole compound according to claim 1 in the preparation of a drug resistance reversal agent for human oral epidermoid carcinoma cells, characterized in that: The β-carboline-thiazole compound is any one of the following structures: 。

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