Beta-carboline-thiazole compounds, methods of making and use as resistance reversal agents

Abstract

The application discloses a beta-carboline-thiazole compound, a preparation method thereof and application of the beta-carboline-thiazole compound as a drug resistance reversing agent, and belongs to the technical field of medicinal chemistry. The structure of the beta-carboline-thiazole compound is shown in the following formula, the beta-carboline-thiazole compound is synthesized by taking Boc-D-tryptophan as a substrate, removing a protecting group, Pictet-Spengler cascade reaction, aromatization, intermolecular polycondensation and the like. The beta-carboline-thiazole compound has the advantages of novel structure, good safety and higher drug resistance reversing activity on human oral epidermoid carcinoma cell strain KBV, the preparation method has the advantages of cheap and easily available raw materials, simple preparation process and suitability for large-scale industrial production.

Description

TECHNICAL FIELD

[0001] The present application relates to a kind of compound and its preparation method and application, specifically to β- carboline-thiazole compound and its preparation method and the application as drug resistance reversal agent, belong to the technical field of pharmaceutical chemistry. BACKGROUND

[0002] Multidrug resistance (MDR) remains one of the key obstacles to effective chemotherapy of cancer. The increased efflux of a variety of chemotherapeutic drugs through the transmembrane ATP-binding cassette (ABC) transporter superfamily is considered to be an important reason for the multidrug resistance of cancer, and the role of P-glycoprotein (P-gp / ABCB1) has been widely proven. The combination of P-gp drug efflux inhibitors with anticancer drugs that are substrates of P-gp transport is considered to be an anticancer treatment strategy to overcome drug resistance by blocking P-gp-mediated drug efflux. In recent years, scientists have conducted extensive research and have screened non-toxic, highly selective and effective P-gp efflux inhibitors.

[0003] β-carboline is an indole alkaloid containing a tricyclic pyrido [3, 4-b] indole structure. Since the first β-carboline was isolated from Sophora flavescens in 1841, the isolation and synthesis of β-carboline and its derivatives have rapidly developed in the following hundreds of years. β-carboline derivatives have become a highly potential lead structure for developing compounds with anticancer activity due to their rich natural sources, high structural diversity, rapid chemical reactivity and interaction ability with various anticancer targets such as DNA intercalation and groove binding, enzymes such as GPX4, topoisomerase, kinase, and proteins such as tubulin, ABCG2 / BCRP, etc.

[0004] Thiazoles have a unique chemical structure and can form a rich variety of derivatives through the introduction and modification of various functional groups. These derivatives exhibit significant activity in interacting with transporters such as P-gp and can effectively inhibit drug efflux, thereby reversing drug resistance.

[0005] Although β-carboline compounds have been widely studied, research on β-carboline-thiazole skeleton compounds, especially those with a thiazole substituent at the C-3 position of β-carboline, is relatively rare, and their drug resistance activity is unknown. SUMMARY

[0006] The present application aims to creatively fuse the two pharmacophores of β-carboline and thiazole, provide a new class of β-carboline-thiazole compounds with a novel skeleton, and the preparation method and drug resistance application of the compounds.

[0007] To achieve the above-mentioned goal, the technical scheme adopted by the present application is as follows:

[0008] β-carboline-thiazole compounds, the structures of which are shown below:

[0009]

[0010] Where R1 can be any of the following structures:

[0011] , , , , , ;

[0012] R2 can be any of the following structures:

[0013] , .

[0014] The aforementioned method for preparing β-carboline-thiazole compounds, wherein R2 of the β-carboline-thiazole compounds is... The preparation method of this β-carboline-thiazole compound includes the following steps:

[0015] Step 1: Dissolve Boc-D-tryptophan in dichloromethane, add pentafluorophenyl diphenylphosphine ester and triethylamine sequentially, stir at room temperature, add azidocysteine ​​dimethyl ester and triphenylphosphine, react at 45°C for 5 h, remove solvent by vacuum rotary evaporation to obtain 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 2 h, after the reaction is complete, quench the reaction, extract with dichloromethane and combine the organic phases, dry the organic phase, remove solvent, and use V... 石油醚 V 乙酸乙酯 After passing the solvent through a column chromatography system with a ratio of 5:1, a yellow oily compound 3 was obtained.

[0016] Step 2: Dissolve compound 3 in dichloromethane, add a benzaldehyde compound or glyoxal dimethyl acetal at room temperature, wherein 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 2 hours, quench the reaction after completion, extract with dichloromethane and combine the organic phases, dry the organic phase, remove the solvent, dissolve the product in tetrahydrofuran, add potassium permanganate, react at room temperature for 3 hours, and then use V 石油醚 V 乙酸乙酯 =3:1 or 2:1 column chromatography, followed by drying to obtain the final product.

[0017] The foregoing method for preparing a β-carboline-thiazole compound, R2 of which is The method for preparing a β-carboline-thiazole compound, R2 of which is

[0018] Step 1: Boc-D-tryptophan was dissolved in dichloromethane, and pentafluorophenyl diphenylphosphinic acid and triethylamine were added in sequence, stirred at room temperature, and azidocysteine dimethyl ester and triphenylphosphine were added, reacted at 45°C for 5h, and the solvent was removed by vacuum evaporation to obtain a thiazoline product. The thiazoline product was dissolved in dichloromethane, and 1,8-diazabicyclo[5,4,0]undec-7-ene and chloroform were added, reacted at room temperature for 2h, and after the reaction was completed, the reaction was quenched, extracted with dichloromethane, and the organic phase was combined and dried, and the solvent was removed, and the product was dissolved in tetrahydrofuran, and potassium permanganate was added, reacted at room temperature for 3h, and the product was separated by column chromatography with V 石油醚 :V 乙酸乙酯 =5:1, and the solvent was removed to obtain compound 3 in the form of yellow oil;

[0019] Step 2: Compound 3 was dissolved in dichloromethane, and a benzaldehyde compound or glyoxal dimethyl acetal was added at room temperature, the benzaldehyde compound being any one of p-methoxybenzaldehyde, trimethoxybenzaldehyde, p-dimethylaminobenzaldehyde, 3-trifluoromethylbenzaldehyde and 4-cyanobenzaldehyde, and trifluoroacetic acid was added dropwise at 0°C, and the reaction was carried out at room temperature for 2h, and after the reaction was completed, the reaction was quenched, extracted with dichloromethane, and the organic phase was combined and dried, and the solvent was removed, and the product was dissolved in tetrahydrofuran, and potassium permanganate was added, reacted at room temperature for 3h, and the product was separated by column chromatography with V 石油醚 :V 乙酸乙酯 =3:1 or 2:1, dried to obtain intermediate product A;

[0020] Step 3: Intermediate product A was dissolved in a mixed solution of tetrahydrofuran and methanol, and then aqueous lithium hydroxide solution was added, and the reaction was carried out at room temperature for 10h, and the pH was adjusted to 3.5, and extracted with ethyl acetate and water, and the aqueous phase was back-extracted with ethyl acetate, and the organic phase was combined and dried to obtain intermediate product B;

[0021] Step 4: Intermediate product B was dissolved in dichloromethane, and then 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate, 1-hydroxybenzotriazole and N,N-diisopropylethylamine were added, and the reaction was carried out at room temperature for 10min, and then 5-chlorotryptamine hydrochloride was added, and the reaction was carried out at room temperature for 15h, and extracted with dilute hydrochloric acid and dichloromethane, and the aqueous phase was back-extracted with dichloromethane, and the organic phase was combined and extracted with sodium hydroxide solution, and the aqueous phase was back-extracted with dichloromethane, and the organic phase was combined and dried, and separated by column chromatography with V 石油醚 :V 乙酸乙酯 =4:1, and dried to obtain the product;

[0022] .

[0023] Preferably, in step 1, the molar ratio of Boc-D-tryptophan, pentafluorophenyl diphenylphosphinic acid ester, triethylamine, azidocysteine dimethyl ester, triphenylphosphine, 1,8-diazabicyclo[5,4,0]undec-7-ene and trichlorobromomethane is 4:4:8:1:10:6:6.

[0024] Preferably, in step 2, the molar ratio of compound 3, dialdehyde dimethyl acetal, p-methoxybenzaldehyde, trimethoxybenzaldehyde, p-dimethylaminobenzaldehyde, 3-trifluoromethylbenzaldehyde or 4-cyanobenzaldehyde is 2:3.

[0025] Preferably, in step 3, the molar ratio of intermediate product A and lithium hydroxide is 2:15.

[0026] Preferably, in step 4, the molar ratio of intermediate product B, 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate, 1-hydroxybenzotriazole, N,N-diisopropylethylamine and 5-chloro tryptamine hydrochloride is 20:26:1:72:30.

[0027] The foregoing β-carboline-thiazole compound is used for preparing a human oral epidermoid carcinoma cell drug resistance reversing agent, wherein the β-carboline-thiazole compound is any one of the following structures:

[0028]

[0029] The present application has the advantages of:

[0030] (1) The β-carboline-thiazole compound provided by the present application has a novel structure, good safety and higher drug resistance reversing activity on human oral epidermoid carcinoma cell strain KBV than verapamil.

[0031] (2) The preparation method of the β-carboline-thiazole compound provided by the present application has the advantages of cheap and easily available raw materials, simple preparation process and suitability for large-scale industrial production. BRIEF DESCRIPTION OF DRAWINGS

[0032] Figure 1 is the IC value evaluation graph of paclitaxel alone on drug-resistant human oral cancer cell strain KBV. 50

[0033] Figure 2 is the IC value evaluation graph of paclitaxel on drug-resistant human oral cancer cell strain KBV under the condition of co-administration of verapamil and paclitaxel. 50 ​​

[0034] Figure 3 is the IC value evaluation chart of paclitaxel on drug-resistant oral cancer cell strain KBV under the condition of co-administration of compound 4 and paclitaxel. 50

[0035] Figure 4 is the IC value evaluation chart of paclitaxel on drug-resistant oral cancer cell strain KBV under the condition of co-administration of compound 5 and paclitaxel. 50

[0036] Figure 5 is the IC value evaluation chart of paclitaxel on drug-resistant oral cancer cell strain KBV under the condition of co-administration of compound 9 and paclitaxel. 50

[0037] Figure 6 is the IC value evaluation chart of paclitaxel on drug-resistant oral cancer cell strain KBV under the condition of co-administration of compound 11 and paclitaxel. 50 DETAILED DESCRIPTION

[0038] The application will be specifically introduced below in combination with specific examples.

[0039] I. Structure of β-carboline-thiazole compound

[0040] The β-carboline-thiazole compound provided by the application is structurally shown as follows:

[0041]

[0042] In the formula, R1 is any one of the following structures:

[0043]

[0044] R2 is any one of the following structures:

[0045]

[0046] II. Preparation method of β-carboline-thiazole compound

[0047] The preparation method of the above β-carboline-thiazole compound provided by the application is as follows: taking Boc-D-tryptophan as a substrate, the synthesis of the above β-carboline-thiazole compound is completed through the steps of removal of a protecting group, Pictet-Spengler cascade reaction, aromatization, and intermolecular polycondensation. ​​​​​​​​​​​​

[0048] Example 1

[0049]

[0050] Step 1: Dissolve commercially available Boc-D-tryptophan (Compound 1, 1 mmol) in dry dichloromethane (DCM, 50 mL), add five fluorophenyl diphenylphosphinate (FDPP, 1 mmol) and triethylamine (Et3N, 2 mmol) successively, stir at room temperature for 5 min, then add azidocysteine dimethyl ester (Compound 2, 0.25 mmol) and triphenylphosphine (PPh3, 2.5 mmol), react at 45 °C for 5 h, remove the solvent by vacuum rotary evaporation, to obtain a thiazoline product, without purification, dissolve the obtained thiazoline product in dry DCM (50 mL), add 1,8-diazabicyclo[5,4,0]undec-7-ene (DBU, 1.5 mmol) and trichlorobromomethane (CBrCl3, 1.5 mmol), react at room temperature for 2 h, monitor the completion of the reaction by thin layer chromatography (TLC), then add saturated ammonium chloride solution (80 mL) to the reaction system to quench the reaction, extract with dichloromethane (80 mL each time, extract 3 times) and combine the organic phases, dry the organic phase with anhydrous sodium sulfate, remove the solvent by vacuum rotary evaporation, and purify by column chromatography with V 石油醚 :V 乙酸乙酯 = 5:1, remove the solvent by vacuum rotary evaporation, to obtain a yellow oil (Compound 3) with a yield of 79%.

[0051] The data of H NMR and ESI-HRMS of Compound 3 are as follows: 1

[0052] 1 H NMR (400 MHz, 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= 7.6 Hz, J= 7.6 Hz, 1H), 7.08 (dd, J= 7.6 Hz, J= 7.6 Hz, 1H), 6.69 (d, J= 0.8 Hz, 1H), 5.39 (s, br, 2H), 3.98 (s, 3H), 3.55 (d, J= 13.2 Hz, 1H), 3.45 (dd, J= 4.0 Hz, J= 13.2 Hz, 1H), 1.28 (s, 9H);

[0053] ​ESI-HRMS: C 20 H 23 N3O4S ([M+Na)) + Calculated value: 424.1302; theoretical value: 423.2308.

[0054] Step 2: Compound 3 (1 mmol) was dissolved in dry DCM (20 mL), and p-methoxybenzaldehyde (1.5 mmol) was added at room temperature. Trifluoroacetic acid (TFA, 1 mL) was added dropwise at 0 °C. The reaction was allowed to proceed at room temperature for 2 h. After the reaction was completed by TLC monitoring, a saturated sodium bicarbonate solution (50 mL) was added to the reaction system to quench the reaction. The mixture was extracted with dichloromethane (40 mL each time, 3 extractions) 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 tetrahydrofuran (THF, 30 mL), and potassium permanganate (KMnO4, 1.5 mmol) was added. The reaction was allowed to proceed at room temperature for 3 h. The product was then analyzed by V... 石油醚 V 乙酸乙酯 The mixture was passed through a column chromatography system at a ratio of 3:1 and dried to give a pale yellow solid (compound 4) with a yield of 75%.

[0055] Compound 4 1 H NMR, 13 The specific data for C NMR and ESI-HRMS are as follows:

[0056] 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);

[0057] 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;

[0058] ESI-HRMS: C 23 H 17N3NaO3S ([M+Na] + Calculated 438.0883, found 438.0880.

[0059] Example 2

[0060]

[0061] Step 1: exactly the same as step 1 of example 1, no further description.

[0062] 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, the reaction was carried out at room temperature for 2 h, TLC monitoring showed that the reaction was completed, saturated sodium bicarbonate solution (50 mL) was added to the reaction system to quench the reaction, dichloromethane was used for extraction (40 mL each time, extracted 3 times) and the organic phase was combined, anhydrous sodium sulfate was used to dry the organic phase, the solvent was removed by vacuum evaporation, the product was dissolved in dry THF (30 mL), KMnO4 (1.5 mmol) was added, the reaction was carried out at room temperature for 3 h, the column was dried after V 石油醚 =3:1, dried to obtain a light yellow solid (compound 5) with a yield of 67%. 乙酸乙酯

[0063] The data of H NMR, C NMR and ESI-HRMS of compound 5 are as follows: 1 H NMR, 13 C NMR and ESI-HRMS of compound 5 are as follows:

[0064] 1 H NMR (500 MHz, CDCl3) δ 9.20 (s, 1H), 8.85 (s, 1H), 8.19 (s, 1H), 8.09 (d, J =8.0 Hz, 1H), 7.57 (d, J =5.0 Hz, 2H), 7.34-7.31 (m, 1H), 7.18 (s, 2H), 4.00 (s, 3H), 3.90 (s, 3H), 3.89 (s, 6H);

[0065] 13 C NMR (126 MHz, CDCl3) δ 172.0, 162.5, 153.9, 147.4, 142.4, 141.2, 140.9, 139.0, 134.5, 133.5, 130.8, 129.1, 128.9, 122.4, 122.4, 121.1, 112.1, 111.0, 105.8, 61.1, 56.5, 52.6, 29.8;

[0066] ​ESI-HRMS: C 25 H 21 N3NaO5S ([M+Na] + ) calc. 498.1094, found 498.1089.

[0067] Example 3

[0068]

[0069] Step 1: exactly the same as step 1 of example 1, no further description.

[0070] 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, the reaction was carried out at room temperature for 2 h, TLC monitoring showed that the reaction was completed, saturated sodium bicarbonate solution (50 mL) was added to the reaction system to quench the reaction, dichloromethane was used for extraction (40 mL each time, extracted for 3 times) and the organic phase was combined, anhydrous sodium sulfate was used to dry the organic phase, the solvent was removed by vacuum rotary evaporation, the product was dissolved in dry THF (30 mL), KMnO4 (1.5 mmol) was added, the reaction was carried out at room temperature for 3 h, V 石油醚 :V 乙酸乙酯 =2:1 over the column, dried to obtain a light yellow solid (compound 6), yield 65%.

[0071] The data of compound 6 were as follows: 1 H NMR, 13 C NMR and ESI-HRMS:

[0072] 1 H NMR (400 MHz, CDCl3) δ 8.82 (d, J =10.0 Hz, 2H), 8.18 (s, 1H), 8.07 (d, J =7.6 Hz, 1H), 7.90 (d, J =8.8 Hz, 2H), 7.56-7.47 (m, 2H), 7.28 (m, 1H), 6.83 (d, J =8.8 Hz, 2H), 3.98 (s, 3H), 3.00 (s, 6H);

[0073] 13C NMR (100 MHz, CDC13) 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;

[0074] ESI-HRMS: C 24 H 20 N4NaO2S ([M+Na] + ) calcd 451.1199, found 451.1200.

[0075] Example 4

[0076]

[0077] Step 1: exactly the same as step 1 of example 1, no further description.

[0078] 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, the reaction was carried out at room temperature for 2 h, after TLC monitoring the reaction was completed, saturated sodium bicarbonate solution (50 mL) was added to the reaction system to quench the reaction, extracted with dichloromethane (40 mL each time, extracted 3 times) and the organic phase was combined, anhydrous sodium sulfate was used to dry the organic phase, the solvent was removed by vacuum rotary evaporation, the product was dissolved in dry THF (30 mL), KMnO4(1.5 mmol) was added, the reaction was carried out at room temperature for 3 h, the column was passed through with V 石油醚 :V 乙酸乙酯 =2:1, dried to obtain a light yellow solid (compound 7), yield 67%.

[0079] The data of 1 H NMR, 13 C NMR and ESI-HRMS of compound 7 are as follows:

[0080] 1 H NMR (500 MHz, CDC13) δ 9.32 (s, 1H), 8.97 (s, 1H), 8.21 (s, 1H), 8.15 (d, J =8.0 Hz, 1H), 7.57-7.52 (m, 2H), 7.29 (t, J =6.5 Hz, 1H), 5.69 (s, 1H), 4.00 (s, 3H), 3.56 (s, 6H);

[0081] 13C NMR (125 MHz, CDC13) 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;

[0082] ESI-HRMS: C 19 H 17 N3NaO4S ([M+Na] + ) calcd 406.0832, found 406.0830.

[0083] Example 5

[0084]

[0085] Step 1: exactly the same as step 1 of example 1, no further description.

[0086] 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, the reaction was carried out at room temperature for 2 h, TLC monitoring reaction was completed, saturated sodium bicarbonate solution (50 mL) was added to the reaction system to quench the reaction, extracted with dichloromethane (40 mL each time, extracted 3 times) and the organic phase was combined, anhydrous sodium sulfate was used to dry the organic phase, the solvent was removed by vacuum evaporation, the product was dissolved in dry THF (30 mL), KMnO4 (1.5 mmol) was added, the reaction was carried out at room temperature for 3 h, the column was passed through V 石油醚 :V 乙酸乙酯 =2:1, dried to obtain a light yellow solid (compound 8), yield 74%.

[0087] The data of 1 H NMR, 13 C NMR and ESI-HRMS of compound 8 are as follows:

[0088] 1 H NMR (500 MHz, d -DMSO) δ 11.96 (s, 1H), 8.87 (s, 1H), 8.48 (s, 1H), 8.44 (d, J =8 Hz, 1H), 8.37 (d, J =7.5 Hz, 2H), 7.93 (d, J =8 Hz, 1H), 7.88 (t, J =8 Hz, 1H), 7.68 (d,J = 8 Hz, 1H), 7.61 (t, J = 8 Hz, 1H), 7.32 (t, J = 7.5 Hz, 1H), 2.53 (s, 3H);

[0089] 13 C NMR (125 MHz, DMSO) 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;

[0090] ESI-HRMS: C 23 H 14 F3N3NaO2S ([M+Na] + ) calcd 476.0651, found 476.0649.

[0091] Example 6

[0092]

[0093] Step 1: exactly the same as step 1 of example 1, no further description.

[0094] 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, the reaction was carried out at room temperature for 2 h, TLC monitoring showed that the reaction was completed, saturated sodium bicarbonate solution (50 mL) was added to the reaction system to quench the reaction, extracted with dichloromethane (40 mL each time, extracted 3 times) and the organic phase was combined, anhydrous sodium sulfate was used to dry the organic phase, the solvent was removed by vacuum evaporation, the product was dissolved in dry THF (30 mL), KMnO4 (1.5 mmol) was added, the reaction was carried out at room temperature for 3 h, the column was eluted with V 石油醚 :V 乙酸乙酯 = 2: 1, dried to obtain a light yellow solid (compound 9), yield 75%.

[0095] The data of 1 H NMR, 13 C NMR and ESI-HRMS of compound 9 are as follows:

[0096] 1 H NMR (500 MHz,d- DMSO) 11.92 (s, 1H), 8.88 (s, 1H), 8.49 (s, 1H), 8.44 (d, J = 8.0 Hz, 1H), 8.22 (d, J = 8.0 Hz, 1H), 8.08 (d, J = 8.0 Hz, 2H), 7.64 (d, J = 8.5 Hz, 2H), 7.59 (t, J = 8.0 Hz, 1H), 7.48 (d, J = 8.0 Hz, 1H), 7.29 (t, J = 7.5 Hz, 1H), 3.89 (s, 3H); J J J J J J = 7.5 Hz, 1H), 3.89 (s, 3H);

[0097] 13 C NMR (125 MHz, DMSO) δ d- 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;

[0098] ESI-HRMS: C 23 H 14 N4NaO2S ([M+Na] + ) calcd 433.0728, found 433.0730.

[0099] Example 7

[0100]

[0101] Step 1: Compound 4 prepared in Example 1 (1 mmol) was dissolved in a mixture of THF and methanol (MeOH) (V THF :V MeOH = 1:1), then lithium hydroxide (LiOH) aqueous solution (concentration 0.5 mmol / mL, 15 mL) was added, and the reaction was carried out at room temperature for 10 h. After TLC detection, the reaction was complete, the pH was adjusted to 3.5 using dilute hydrochloric acid (concentration 1 M), then extracted with ethyl acetate and water, the water phase was back-extracted twice with ethyl acetate, the organic phase was combined and rotary evaporated to obtain compound 10, which was used directly in the next step without purification.

[0102] ​​​​​​Step 2: Compound 10 (1 mmol) was dissolved in dry DCM (50 mL), followed by the addition of 2-(7-azabenzotriazol-1 -yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU, 1.3 mmol), 1 -hydroxybenzotriazole (HOBt, 0.05 mmol) and N,N-diisopropylethylamine (DIPEA, 3.6 mmol), after 10 min at room temperature, 5-chlorotryptamine hydrochloride (1.5 mmol) was added, the reaction was left to proceed for 15 h at room temperature, after TLC monitoring of the complete reaction, the mixture was extracted with diluted hydrochloric acid (0.3 M concentration) and dichloromethane, the aqueous phase was back-extracted twice with dichloromethane, the organic phases were combined and extracted with sodium hydroxide solution (0.5 M concentration), the aqueous phase was back-extracted twice with dichloromethane, the organic phases were combined, dried and evaporated to dryness, the residue was taken up in V 石油醚 :V 乙酸乙酯 = 4:1 over a column, dried, obtaining a white solid (compound 11) with a yield of 83%.

[0103] The data of H NMR, C NMR and ESI-HRMS of compound 11 are as follows: 1 H NMR, 13 C NMR and ESI-HRMS of compound 11 are as follows:

[0104] 1 H NMR (500 MHz, CDCI3) δ 8.71 (s, 1 H), 8.63 (s, 1 H), 8.22 (s, 1 H), 8.21 - 8.19 (m, 1 H), 8.13 (s, 1 H), 8.03 - 7.97 (d, 1 H), 7.73 (t, J = 6.2 Hz, 1 H), 7.67 (d, J = 2.0 Hz, 1 H), 7.63 - 7.53 (m, 2 H), 7.37 (t, J = 7.3 Hz, 1 H), 7.32 (d, J = 8.6 Hz, 1 H), 7.15 (ddd, J = 8.5, 4.7, 2.2 Hz, 4 H), 3.92 (s, 3 H), 3.84 (q, J = 6.8 Hz, 3 H), 3.13 (t, J = 7.0 Hz, 2 H);

[0105] 13C NMR (125 MHz, CDC13) 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, 123.45, 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;

[0106] ESI-HRMS: C 32 H 24 ClN5O2S ([M+Na] + ) calcd 577.1335, found 577.1339.

[0107] III. Pharmacological test of β-carboline-thiazole compounds

[0108] The drug resistance reversal activity of compound 4, compound 5, compound 6, compound 7, compound 8, compound 9 and compound 11 on drug-resistant oral cancer cell line KBV was detected.

[0109] The effect of the above compounds on the survival rate of drug-resistant oral epidermoid carcinoma cell line KBV was determined by MTT method in the case of separate administration.

[0110] The effect of the above compounds on the survival rate of drug-resistant oral epidermoid carcinoma cell line KBV and the half inhibitory concentration (IC 50 ) were determined by MTT method in the case of co-administration with paclitaxel (PTX).

[0111] Detection principle: 3-(4,5-dimethylthiazole-2)-2,5-diphenyl tetrazolium bromide (MTT) can penetrate the cell membrane into the cell, and succinate dehydrogenase in the mitochondria of living cells can reduce the exogenous MTT to a blue-purple needle-shaped formazan crystal which is difficult to dissolve in water and is deposited in the cell. The crystal can be dissolved by dimethyl sulfoxide (DMSO), and the determination of its absorbance can reflect the cell survival rate. The half inhibitory concentration (IC 50 ) is defined as the drug concentration when 50% of tumor cells survive. According to the measured optical density (OD value), a standard curve of cell survival rate is made, and then the corresponding drug concentration is obtained on the curve.

[0112] The determination method is as follows:

[0113] (1) Drug-resistant oral epidermoid carcinoma cell line KBV was inoculated in a 96-well plate at a density of 3000 cells / well, and cultured in a CO2 incubator for 24 h;

[0114] (2) PTX (10 mM stock solution) was diluted to 1000 nM, 300 nM, 100 nM, 30 nM, 10 nM, 3 nM, 1 nM, 0.3 nM, 0.1 nM, and the concentrations of compound 4, compound 5, compound 6, compound 7, compound 8, compound 9 and compound 11 (10 mM stock solution) were diluted to 10 μM, respectively, and the concentration of verapamil (control compound, 10 mM stock solution) was diluted to 10 μM;

[0115] (3) Three groups of parallel holes were set for each concentration of PTX, and different concentrations of PTX were added to the inoculated cells alone (for calculating the IC 50 of PTX), or different concentrations of PTX and 10 μM of the test compound were added to the inoculated cells together and equally (for calculating the cell survival rate and the IC 50 of PTX under the condition of co-administration), or 10 μM of the test compound was added to the inoculated cells alone (for calculating the cell survival rate under the condition of single administration), and the incubation was continued for 72 h;

[0116] (4) The 96-well plate was taken out from the CO2incubator, 30 μL of MTT solution (5 mg / mL) was added to each well, and the incubation was continued in the CO2incubator for 4 h;

[0117] (5) The upper liquid was discarded, 150 μL of DMSO was added to each well, and the crystal of formazan was completely dissolved by gently shaking for 20 min;

[0118] (6) The absorbance of each well was measured at 490 nm by using the enzyme label instrument, and the cell survival rate was calculated.

[0119] The experimental results were averaged from three parallel experiments.

[0120] The survival rates of the drug-resistant oral epidermoid carcinoma cell line KBV under the condition of single administration and co-administration were measured and are shown in Table 1.

[0121] Table 1 Survival rates of the drug-resistant oral epidermoid carcinoma cell line KBV

[0122]

[0123] From Table 1, it can be seen that:

[0124] (1) Under the condition of single administration, except for compound 7 and compound 9 (the survival rate of the drug-resistant oral epidermoid carcinoma cell line KBV was less than 80%), the other compounds showed good safety;

[0125] (2) Compound 4, Compound 5, Compound 9 and Compound 11 all showed significant inhibitory effect on drug-resistant KBV oral epidermoid carcinoma cell line, and the cell survival rate was less than 37% when they were co-administered with PTX.

[0126] The compounds 4, 5, 9 and 11 with excellent co-administration effect were selected, and verapamil was used as a control to evaluate the half inhibitory concentration (IC 50 ) of PTX under the condition of co-administration of these compounds and PTX.

[0127] The IC 50 value of PTX on drug-resistant KBV oral cancer cell line under the condition of single administration of PTX is shown in Figure 1 .

[0128] The IC 50 value of PTX on drug-resistant KBV oral cancer cell line under the condition of co-administration of verapamil and PTX is shown in Figure 2 .

[0129] The IC 50 value of PTX on drug-resistant KBV oral cancer cell line under the condition of co-administration of Compound 4 and PTX is shown in Figure 3 .

[0130] The IC 50 value of PTX on drug-resistant KBV oral cancer cell line under the condition of co-administration of Compound 5 and PTX is shown in Figure 4 .

[0131] The IC 50 value of PTX on drug-resistant KBV oral cancer cell line under the condition of co-administration of Compound 9 and PTX is shown in Figure 5 .

[0132] The IC 50 value of PTX on drug-resistant KBV oral cancer cell line under the condition of co-administration of Compound 11 and PTX is shown in Figure 6 .

[0133] The IC 50 value of PTX on drug-resistant KBV oral cancer cell line under the condition of single administration of PTX can be obtained from Figure 1 . The IC 50 value of PTX on drug-resistant KBV oral cancer cell line under the condition of co-administration of each test compound and PTX can be obtained from Figures 2 to 6 .

[0134] The IC 50 value of PTX on drug-resistant KBV oral cancer cell line is shown in Table 2.

[0135] Table 2 IC50 of PTX under co-administration conditions 50 value

[0136]

[0137] As shown in Table 2:

[0138] (1) IC50 of PTX alone against drug-resistant human oral epidermal carcinoma cell line KBV 50 The value is as high as 1612.0 nM;

[0139] (2) When PTX was co-administered with verapamil, compound 4, compound 5, compound 9, and compound 11, the IC50 of PTX against the drug-resistant human oral epidermal carcinoma cell line KBV was significantly reduced. 50 The values ​​were significantly reduced. Among them, compounds 4, 5, 9 and 11 all showed significant resistance reversal activity against paclitaxel compared to the commercially available verapamil. In particular, compounds 5 and 9 showed resistance reversal activity that was more than 3 times higher than that of the commercially available drug verapamil, and 89.5 times higher than that of paclitaxel alone.

[0140] Considering that compound 9 still has a certain impact on the survival rate of drug-resistant human oral epidermal carcinoma cell line KBV under high concentration alone, and has cytotoxicity, compound 5 has a better application prospect than compound 9.

[0141] It should be noted that the above embodiments are merely examples for clearly illustrating the present invention, and are not intended to limit the implementation of the present invention. Those skilled in the art can make other variations or modifications based on the above description. It is impossible to exhaustively list all possible implementations here. All obvious variations or modifications derived from the technical solutions of this invention are still within the scope of protection of this invention.

Claims

1. A β-carboline-thiazole compound characterized in that, The structure is shown as follows: 。 2. The process for the preparation of the β-carboline-thiazole compounds according to claim 1, characterized in that, comprising the following steps: Step 1: Dissolve Boc-D-tryptophan in dichloromethane, add five-fluorophenyl diphenylphosphinic acid and triethylamine in sequence, stir at room temperature, add azidocysteine dimethyl ester and triphenylphosphine, react at 45°C for 5h, remove the solvent by vacuum evaporation to obtain thiazoline product, dissolve the thiazoline product in dichloromethane, add 1,8-diazabicyclo[5,4,0]undec-7-ene and chloroformic bromide, react at room temperature for 2h, after the reaction is completed, quench the reaction, extract with dichloromethane and combine the organic phase, dry the organic phase, remove the solvent, pass through a column with V 石油醚 :V 乙酸乙酯 =5:1, remove the solvent to obtain compound 3 as yellow oil; Step 2: Compound 3 is dissolved in dichloromethane, p-methoxybenzaldehyde is added at room temperature, trifluoroacetic acid is added dropwise at 0°C, the reaction is allowed to proceed at room temperature for 2 h, after the reaction is complete, the reaction is quenched, extracted with dichloromethane and the organic phases are combined, the organic phase is dried, the solvent is removed, the product is dissolved in tetrahydrofuran, potassium permanganate is added, the reaction is allowed to proceed at room temperature for 3 h, V 石油醚 :V 乙酸乙酯 = 3: 1 or 2: 1, dried, to give compound 4 as a pale yellow solid; Step 3: Compound 4 was dissolved in a mixed solution of tetrahydrofuran and methanol, then lithium hydroxide aqueous solution was added, and the reaction was carried out at room temperature for 10 h, the pH was adjusted to 3.5, extracted with ethyl acetate and water, the aqueous phase was back-extracted with ethyl acetate, the organic phases were combined and rotary evaporated to obtain compound 10; Step 4: Compound 10 is dissolved in dichloromethane, followed by the addition of 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate, 1-hydroxybenzotriazole and N,N-diisopropylethylamine, after 10 min at room temperature 5-chlorotryptamine hydrochloride is added, the reaction is continued for 15 h at room temperature, the mixture is extracted with dilute hydrochloric acid and dichloromethane, the aqueous phase is back-extracted with dichloromethane, the combined organic phases are extracted with a sodium hydroxide solution, the aqueous phase is back-extracted with dichloromethane, the combined organic phases are dried and evaporated, the residue is dissolved in dichloromethane and purified by column chromatography (V 石油醚 : V 乙酸乙酯 = 4:1) and dried. Step 5: Compound 11 is dissolved in dichloromethane, followed by the addition of 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate, 1-hydroxybenzotriazole and N,N-diisopropylethylamine, after 10 min at room temperature 5-chlorotryptamine hydrochloride is added, the reaction is continued for 15 h at room temperature, the mixture is extracted with dilute hydrochloric acid and dichloromethane, the aqueous phase is back-extracted with dichloromethane, the combined organic phases are extracted with a sodium hydroxide solution, the aqueous phase is back-extracted with dichloromethane, the combined organic phases are dried and evaporated, the residue is dissolved in dichloromethane and purified by column chromatography (V 石油醚 : V 乙酸乙酯 = 4:1) and dried. 。 3. The preparation method according to claim 2, characterized in that, In step 1, the molar ratio of Boc-D-tryptophan, pentafluorophenyl diphenyl phosphinic acid ester, triethylamine, azidocysteine dimethyl ester, triphenyl phosphine, 1,8-diazabicyclo[5,4,0]undec-7-ene and trichlorobromomethane is 4:4:8:1:10:6:

6.

4. The preparation method according to claim 2, characterized in that, In step 2, the molar ratio of compound 3 and p-methoxybenzaldehyde is 2:

3.

5. The preparation method according to claim 2, characterized in that, In step 3, the molar ratio of compound 4 and lithium hydroxide is 2:

15.

6. The preparation method according to claim 2, characterized in that, In step 4, the molar ratio of compound 10, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, 1-hydroxybenzotriazole, N,N-diisopropylethylamine and 5-chloro tryptamine hydrochloride is 20:26:1:72:

30.

7. Use of β-carboline-thiazole compounds 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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