Lasiokaurine-tetrazole sulfide derivatives, methods of making and using the same

Abstract

The application discloses a kind of two-faced needle alkali-tetrazole sulfide derivatives, the derivative has as general formula I or general formula II structure: The application provides a kind of two-faced needle alkali-tetrazole sulfide derivatives, its structure is novel, and substrate applicability is wide, easy to operate, byproduct is less, it is convenient to separate and purify, can be applicable to larger scale preparation, has considerable application prospect.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of organic synthesis. More particularly, the present application relates to a nitidine-tetrazole sulfide derivative and a preparation method and application thereof. BACKGROUND

[0002] At present, traditional drugs have the disadvantages of poor water solubility, low dissolution rate, low permeability, etc., resulting in poor drug efficacy. Moreover, most drugs for treating cancer have side effects, such as cisplatin having nephrotoxicity, peripheral neurotoxicity and other side effects. Therefore, exploring new active ingredients from plants which are significantly effective for human diseases, and modifying and reforming the active ingredients to enhance efficacy and reduce toxicity are one of the research focuses of chemical researchers at home and abroad in recent years

[0003] Nitidine is a very promising traditional Chinese medicine for preventing and treating tumors. Nitidine chloride (NC) is a phenanthridine alkaloid component isolated from nitidine, which has a wide range of pharmacological effects, such as anti-inflammatory, antibacterial, analgesic, antitumor, antifungal activities, etc. In recent years, it has been found that NC has good antitumor activity and has a strong inhibitory effect on various cancer cells. Due to the low content of NC in plants, poor solubility, complicated extraction and purification process, non-specific toxicity to normal tissues, and low bioavailability, it hinders its clinical application. Overcoming these defects through structural modification to improve the drug properties is a reasonable strategy to further promote the research of nitidine and its derivatives.

[0004] Studies have shown that tetrazole sulfide derivatives have antitumor activity. Tetrazole sulfide compounds are unsaturated five-membered heterocyclic compounds with four nitrogen atoms on the ring. A large number of studies have shown that they have a wide range of biological activities. In the development of new anticancer drugs, it is considered a promising strategy to combine the tetrazole sulfide fragment with other pharmacophores. Therefore, if some superior antitumor activity fragments of tetrazole sulfide are spliced on the skeleton of nitidine analogs, it is possible to obtain highly efficient and low-toxicity nitidine derivatives, which will further promote the clinical application of nitidine derivatives. SUMMARY

[0005] It is an object of the present application to solve at least the above problems and to provide at least the advantages described later.

[0006] To achieve these objects and other advantages in accordance with the present application, a nitidine-tetrazole sulfide derivative is provided, which has a structure as shown in general formula I or general formula II:

[0007]

[0008] When R1 is methyl, R2 is hydrogen, and R3 is any one of methyl, phenyl, ethylbenzene, chlorobenzene, and methoxybenzene;

[0009] When R1 is hydrogen or R2 is methyl, and R3 is any one of methyl, phenyl, ethylbenzene, chlorobenzene, and methoxybenzene;

[0010] When R1 and R2 are methoxy, and R3 is any one of methyl, phenyl, ethylbenzene, chlorobenzene, and methoxybenzene;

[0011] When R1 and R2 are chlorine, and R3 is any one of methyl, phenyl, ethylbenzene, chlorobenzene, and methoxybenzene.

[0012] The application also provides a preparation method of the epi-two-flank needle alkali-tetrazole sulfide derivative, which comprises the following steps:

[0013] Step one, synthesizing a compound 2 shown in formula 2 by using an acyl chloride compound 1 shown in formula 1 and methoxyamine hydrochloride as raw materials, under the action of potassium carbonate and in an ethyl acetate solvent;

[0014]

[0015] Step two, synthesizing a compound 3 shown in formula 3 by dissolving the compound 2, iodobenzene, palladium acetate and silver oxide in acetic acid, and reacting under nitrogen protection;

[0016]

[0017] Step three, synthesizing a compound 4 shown in formula 4 by dissolving the compound 3 in anhydrous methanol and reacting under ultraviolet lamp irradiation;

[0018]

[0019] Step four, synthesizing a compound 5 shown in formula 5 or a compound 6 shown in formula 6 by using the compound 4 and 1,4-dibromobutane or 1,6-dibromohexane as raw materials, under the action of sodium hydride and in a DMF solvent;

[0020]

[0021] Step five, synthesizing a compound 7 shown in formula 7 by using an isothiocyanic acid compound and sodium azide as raw materials, under the action of zinc chloride and in an acetonitrile solvent;

[0022]

[0023] Step six, synthesis of compound 8 as shown in formula 8 or compound 9 as shown in formula 9, i.e. daphne-tetrazole sulfide derivative of general formula I or general formula II, from compound 5 or compound 6 and compound 7 in acetonitrile solvent with triethylamine;

[0024]

[0025]

[0026] wherein, when R1 is methyl, R2 is hydrogen, R3 is any one of methyl, phenyl, ethylbenzene, chlorobenzene, methoxybenzene;

[0027] when R1 is hydrogen or, R2 is methyl, R3 is any one of methyl, phenyl, ethylbenzene, chlorobenzene, methoxybenzene;

[0028] when R1, R2 is methoxy, R3 is any one of methyl, phenyl, ethylbenzene, chlorobenzene, methoxybenzene;

[0029] when R1, R2 is chlorine, R3 is any one of methyl, phenyl, ethylbenzene, chlorobenzene, methoxybenzene.

[0030] Preferably, in the preparation method of the daphne-tetrazole sulfide derivative, the molar ratio of methoxyamine hydrochloride, potassium carbonate and compound 1 in step one is 1:2:1.

[0031] Preferably, in the preparation method of the daphne-tetrazole sulfide derivative, the molar ratio of compound 2, iodobenzene, palladium acetate and silver oxide in step two is 1:2:0.05:2.

[0032] Preferably, in the preparation method of the daphne-tetrazole sulfide derivative, the ultraviolet lamp used in step three is a 175W ultraviolet high-pressure mercury lamp.

[0033] Preferably, in the preparation method of the daphne-tetrazole sulfide derivative, the molar ratio of compound 4, sodium hydride and 1,4-dibromobutane or 1,6-dibromohexane in step four is 1:3:5.

[0034] Preferably, in the preparation method of the daphne-tetrazole sulfide derivative, the molar ratio of sodium azide, zinc oxide and allyl isothiocyanate in step five is 1.2:1.2:1.

[0035] Preferably, in the preparation method of the daphne-tetrazole sulfide derivative, the molar ratio of compound 5 or compound 6, compound 7 and triethylamine in step six is 1:1:1.5.

[0036] The application further provides application of the daphnaline-tetrazole sulfide derivative in preparation of a medicine for treating tumors.

[0037]

[0038] When R1 is hydrogen or R2 is methyl, R3 is any one of methyl, phenyl, ethylbenzene, chlorobenzene and methoxybenzene;

[0039] When R1 is hydrogen or R2 is methyl, R3 is any one of methyl, phenyl, ethylbenzene, chlorobenzene and methoxybenzene;

[0040] When R1 and R2 are methoxy, R3 is any one of methyl, phenyl, ethylbenzene, chlorobenzene and methoxybenzene;

[0041] When R1 and R2 are chloro, R3 is any one of methyl, phenyl, ethylbenzene, chlorobenzene and methoxybenzene.

[0042] The application at least has the following beneficial effects:

[0043] 1. The application provides the daphnaline-tetrazole sulfide derivative, which has the advantages of novel structure, wide substrate applicability, simple operation, less by-products, easy separation and purification, and large-scale preparation.

[0044] 2. The application discloses a preparation method of the daphnaline-tetrazole sulfide derivative, which comprises the following steps: taking substituted benzoyl chloride as a starting material, and obtaining a simplified daphnaline phenanthridine ketone skeleton through acylation, palladium-catalyzed ring closure and demethoxylation and the like; then, introducing a mercapto tetrazole active fragment into the skeleton based on the phenanthridine ketone skeleton by combining with a nucleophilic substitution reaction to obtain the daphnaline-tetrazole sulfide derivative; and finally, obtaining the target compound general formula I or general formula II through purification.

[0045] 3. The daphnaline-tetrazole sulfide derivative provided by the application has excellent anti-tumor activity, and the target compounds all have certain anti-tumor effects, wherein the compound 8f has significant inhibitory effect on Hepg2, H460 and HeLa cells, and the inhibition rates are 63.29%±3.74%, 69.40%±1.80% and 51.37%±2.16% respectively at a concentration of 40 μmol / L, which is not only superior to the positive control drug 5-fluorouracil, but also equivalent to chlorinated daphnaline, and the toxicity is lower than that of chlorinated daphnaline. Most of the daphnaline-tetrazole sulfide derivatives have little effect on normal cells LO2.

[0046] Other advantages, objects, and features of the application will be apparent from the following specification, and upon examination of the attached drawings. BRIEF DESCRIPTION OF DRAWINGS

[0047] Figure 1 Structural formula of the noscapine-tetrazole sulfide derivative described in one embodiment of the present application;

[0048] Figure 2 Flow chart of the preparation method of the noscapine-tetrazole sulfide derivative described in another embodiment of the present application. DETAILED DESCRIPTION

[0049] The present application will be further described in conjunction with the drawings and embodiments, so that those skilled in the art can implement the present application according to the description.

[0050] It should be understood that the terms such as "have", "contain", and "include" used herein do not exclude the presence or addition of one or more other elements or combinations thereof.

[0051] It should be noted that the experimental methods described in the following embodiments are all conventional methods, and the reagents and materials, unless otherwise specified, can be obtained commercially.

[0052] In the description of the present application, the orientations or positional relationships indicated by the terms "transverse", "longitudinal", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", etc. are based on the orientations or positional relationships shown in the drawings, and are only for the convenience of describing the present application and simplifying the description, and do not indicate or imply that the devices or elements referred to must have a particular orientation, be constructed and operated in a particular orientation, and therefore cannot be understood as a limitation on the present application.

[0053] <EMBODIMENT 1>

[0054] As shown in Figure 1 The present application provides a noscapine-tetrazole sulfide derivative, which has a structural formula as general formula I or general formula II;

[0055] Figure 1 When R1 is methyl and R2 is hydrogen, R3 is any one of methyl, phenyl, ethylbenzene, chlorobenzene, and methoxybenzene;

[0056] When R1 is hydrogen or R2 is methyl, R3 is any one of methyl, phenyl, ethylbenzene, chlorobenzene, and methoxybenzene;

[0057] When R1, R2 are methoxy, R3 is any one of methyl, phenyl, ethylbenzene, chlorobenzene, methoxybenzene;

[0058] When R1, R2 are methoxy, R3 is any one of methyl, phenyl, ethylbenzene, chlorobenzene, methoxybenzene.

[0059] <Embodiment 2>

[0060] As Figure 2 shown, the application provides a preparation method of a daphnaline-tetrazole sulfide derivative, which specifically comprises the following steps:

[0061] Step one, take 150 mL round-bottom flask, weigh methoxyamine hydrochloride (CH3ONH2·HCl) (0.835 g, 10 mmol) and K2CO3 (2.76 g, 20 mmol) and dissolve them in 60 mL ethyl acetate, then stir them in a 0°C cold well, slowly add 15 mL water, then add 15 mL ethyl acetate solution containing compound 1 (such as compounds 1a-1d: m-methylbenzoyl chloride or p-methylbenzoyl chloride or 3,4-dimethoxybenzoyl chloride or 3,4-dichlorobenzoyl chloride) (10 mmol), stir for 5 min, then stir at room temperature for 5 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 1:1), after the reaction is completed, collect the ethyl acetate layer, extract the water layer with ethyl acetate (3×20 mL), combine the organic layers and dry them with anhydrous Na2SO4, and distill them under reduced pressure to obtain compound 2 (such as compounds 2a-2d);

[0062]

[0063] Step two, under the protection of N2, take 100 mL round-bottom flask, add compound 2 (2a-2d) (8 mmol), iodobenzene (1.79 mL, 16 mmol), palladium acetate (0.334 g, 0.4 mmol), silver oxide (3.708 g, 16 mmol), and dissolve them in 60 mL acetic acid, then stir and reflux at 120°C for 26 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1), after the reaction is completed, add 100 mL ethyl acetate solution, filter off the brown insoluble matter, wash the obtained brown-black filtrate with saturated NaHCO3 until the pH is about 7, then wash it with water, separate and extract the organic phase, dry the organic phase with anhydrous Na2SO4, and concentrate it to obtain the residue, which is separated and purified by column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain compound 3 (3a-3d);

[0064]

[0065] Step three, take 50 mL of a vial, add compound 3 (3a-3d) (1.4 mmol), dissolve in 45 mL of anhydrous methanol, and then place it under a 175W ultraviolet light high-pressure mercury lamp for 8h, TLC monitor the reaction (petroleum ether: ethyl acetate = 1:1), after the reaction is completed, directly reduce pressure distillation to obtain compound 4 (4a-4d);

[0066]

[0067] Step four, take 50 mL of a round-bottom flask, add compound 4 (4a-4d) (0.86 mmol) and sodium hydride (60%, 0.105 g, 2.6 mmol), dissolve in 20 mL of DMF, stir at room temperature for 1h, then add 1,4-dibromobutane or 1,6-dibromohexane (4.3 mmol), stir at room temperature for 6h, TLC monitor the reaction (petroleum ether: ethyl acetate = 2:1), after the reaction is completed, quench with ice water, extract with ethyl acetate (3x10 mL), combine the organic phases and wash with saturated sodium chloride solution, dry over anhydrous Na2SO4 and concentrate, the residue is separated and purified by column chromatography (eluent petroleum ether: ethyl acetate = 5:1) to compound 5 (5a-5d) or 6 (6a-6d);

[0068]

[0069] Step five, add sodium azide (0.786 g, 12.1 mmol) and zinc oxide (ZnCl2) (1.649 g, 12.1 mmol) to 30 mL of acetonitrile, stir the mixture and heat to 80°C, then add substituted phenyl isothiocyanate or methyl isothiocyanate (10.1 mmol). Stir the reaction mixture at 80°C for 1h, TLC monitor the reaction (petroleum ether: ethyl acetate = 4:1), after the reaction is completed, remove the solvent by reduced pressure distillation. Stir the residue with 30 mL of 5% (mass fraction) NaOH aqueous solution for 20 min. Filter the suspension, acidify the aqueous layer with concentrated HCl to pH = 1, filter the precipitate formed, wash with water, and dry to obtain the mercaptotetrazole derivative 7 (7a-7e);

[0070]

[0071] Step six, compound 5 (5a-5d) or 6 (6a-6d) and compound 7 (7a-7e) (0.375 mmol) were added to a 25 mL three-necked flask under electromagnetic stirring, dissolved with 10 mL of acetonitrile, heated to 80°C, added triethylamine (0.052 mL, 0.375 mmol) and refluxed for 5 h, TLC monitored the reaction (petroleum ether: ethyl acetate = 2:1), after the reaction was completed, the solvent was removed by reduced pressure distillation, recrystallized with acetone / ethyl acetate to obtain compound 8 (8a-8t) or compound 9 (9a-9t), which is a class of two-azepine-tetrazole sulfide derivatives of general formula I or general formula II;

[0072]

[0073] The specific structural formula and properties of compounds 8a-8t and compounds 9a-9t are as follows:

[0074]

[0075] 8a: white solid, yield 57%, m.p. 147.5-148.6°C; HRMS (ESI) m / z: calcd for C 20 H 21 N5OS[M+H] + : 380.1552, found: 380.1545. 1 H NMR (500 MHz, CDCl3) δ 8.34 (s, 1H), 8.28 (d, J = 7.9 Hz, 1H), 8.18 (d, J = 8.3 Hz, 1H), 7.59 (d, J = 8.1 Hz, 1H), 7.53 (t, J = 7.7 Hz, 1H), 7.38 (d, J = 8.4 Hz, 1H), 7.31 (t, J = 7.6 Hz, 1H), 4.46 (d, J = 7.0 Hz, 2H, -NCH2), 3.90 (s, 3H, -NCH3), 3.45 (t, J = 6.7 Hz, 2H, -SCH2), 2.52 (s, 3H), 2.04–1.97 (m, 4H). 13 C NMR (126 MHz, CDCl3) δ 161.74, 154.36, 138.37, 136.62, 134.08, 131.29, 129.34, 128.70, 125.31, 123.48, 122.63, 121.80, 119.87, 115.05, 41.99, 33.50, 33.04, 27.02, 26.60, 21.51.

[0076]

[0077] 8b: White solid, yield 59%, m.p. 112.3-113.6 °C; HRMS (ESI) m / z: calcd for C 25 H 23 N5OS[M+H] + :442.1701, found:442.1702. 1 H NMR (500 MHz, CDC13) δ 8.33 (s, 1H), 8.27 (d, J = 8.0 Hz, 1H), 8.17 (d, J = 8.3 Hz, 1H), 7.60 - 7.54 (m, 6H), 7.51 (d, J = 7.8 Hz, 1H), 7.38 (d, J = 8.4 Hz, 1H), 7.31 (t, J = 7.5 Hz, 1H), 4.45 (t, J = 7.0 Hz, 2H, -NCH2), 3.50 (t, J = 7.0 Hz, 2H, -SCH2), 2.52 (s, 3H), 2.08 - 2.02 (m, 2H), 1.98 (dt, J = 14.3, 7.0 Hz, 2H). 13 C NMR (126 MHz, CDC13) δ 161.68, 154.41, 138.33, 136.66, 134.03, 133.80, 131.27, 130.24, 129.93, 129.32, 128.70, 125.35, 123.99, 123.47, 122.57, 121.79, 119.84, 115.03, 41.97, 33.05, 26.85, 26.71, 21.51.

[0078]

[0079] 8c: White solid, yield 56%, m.p. 138.1-139.4 °C; HRMS (ESI) m / z: calcd for C 27 H 27 N5OS[M+H] + :470.2014, found:470.2015. 1H NMR (500 MHz, CDC13) δ 8.32 (s, 1H), 8.27 (d, J = 8.0 Hz, 1H), 8.17 (d, J = 8.2 Hz, 1H), 7.58 (d, J = 8.2 Hz, 1H), 7.51 (t, J = 7.7 Hz, 1H), 7.45 (d, J = 8.3 Hz, 2H), 7.37 (t, J = 8.0 Hz, 3H), 7.31 (t, J = 7.5 Hz, 1H), 4.45 (t, J = 6.9 Hz, 2H, -NCH2), 3.48 (t, J = 7.0 Hz, 2H, -SCH2), 2.74 (q, J = 7.5 Hz, 2H), 2.52 (s, 3H), 2.04 (dd, J = 14.4, 7.6 Hz, 2H), 1.98 (dd, J = 14.9, 7.7 Hz, 2H), 1.29 (t, J = 7.6 Hz, 3H). 13 C NMR (126 MHz, CDC13) δ 161.66, 154.38, 146.82, 138.31, 136.69, 134.02, 131.40, 131.27, 129.32, 129.29, 128.71, 125.38, 123.97, 123.46, 122.54, 121.79, 119.84, 115.04, 41.98, 33.01, 28.74, 26.87, 26.71, 21.51, 15.44.

[0080]

[0081] 8d: White solid, yield 62%, m.p. 184.3-185.6 °C; HRMS (ESI) m / z: calcd for C 25 H 22 ClN5OS[M+H] + : 476.1308, found: 476.1312. 1 H NMR (500 MHz, CDC13) δ 8.32 (s, 1H), 8.27 (d, J = 8.0 Hz, 1H), 8.17 (d, J = 8.2 Hz, 1H), 7.58 (d, J = 8.2 Hz, 1H), 7.53 (s, 5H), 7.38 (d, J = 8.4 Hz, 1H), 7.31 (t, J = 7.5 Hz, 1H), 4.46 (t, J = 6.8 Hz, 2H, -NCH2), 3.50 (t, J = 6.9 Hz, 2H, -SCH2), 2.52 (s, 3H), 2.04 (dd, J = 14.3, 7.1 Hz, 2H), 2.01 - 1.95 (m, 2H). 13C NMR (126 MHz, CDC13) δ 161.69, 154.46, 138.35, 136.67, 136.31, 134.05, 132.25, 131.26, 130.19, 129.31, 128.70, 125.36, 125.22, 123.49, 122.57, 121.80, 119.86, 114.99, 41.91, 33.19, 26.82, 26.70, 21.51.

[0082]

[0083] 8e: White solid, yield 60%, m.p. 136.2-138.5 °C; HRMS (ESI) m / z: calcd for C 26 H 25 N5O2S [M+H] + : 472.1802, found: 472.1807. 1 H NMR (500 MHz, CDC13) δ 8.33 (s, 1H), 8.28 (d, J = 7.9 Hz, 1H), 8.17 (d, J = 8.3 Hz, 1H), 7.58 (d, J = 8.2 Hz, 1H), 7.52 (t, J = 7.7 Hz, 1H), 7.44 (d, J = 8.8 Hz, 2H), 7.38 (d, J = 8.4 Hz, 1H), 7.31 (t, J = 7.5 Hz, 1H), 7.03 (d, J = 8.7 Hz, 2H), 4.45 (t, J = 6.6 Hz, 2H, -NCH2), 3.87 (s, 3H, -OCH3), 3.47 (t, J = 6.9 Hz, 2H, -SCH2), 2.52 (s, 3H), 2.07 - 2.01 (m, 2H), 1.98 (dd, J = 14.8, 7.6 Hz, 2H). 13 C NMR (126 MHz, CDC13) δ 161.68, 160.87, 154.56, 138.33, 136.66, 134.04, 131.27, 129.33, 128.70, 126.44, 125.72, 125.35, 123.47, 122.57, 121.79, 119.84, 115.04, 114.99, 55.81, 41.99, 32.96, 26.88, 26.70, 21.52.

[0084]

[0085] 8f: White solid, yield 62%, m.p. 108.2-110.3 °C; HRMS (ESI) m / z: calcd for C20 H 21 N5OS[M+H] + :380.1544, found:380.1545. 1 H NMR (500 MHz, CDC13 ) δ 8.41 (d, J = 8.1 Hz, 1H), 8.30 (d, J = 7.8 Hz, 1H), 8.07 (s, 1H), 7.54 (t, J = 7.4 Hz, 1H), 7.41 (d, J = 8.2 Hz, 1H), 7.38 (d, J = 8.4 Hz, 1H), 7.31 (t, J = 7.5 Hz, 1H), 4.43 (t, J = 6.5 Hz, 2H, -NCH2), 3.90 (s, 3H, -NCH3), 3.44 (t, J = 6.8 Hz, 2H, -SCH2), 2.57 (s, 3H), 2.05 - 1.97 (m, 4H). 13 C NMR (126 MHz, CDC13 ) δ 161.63, 154.34, 143.18, 137.20, 133.71, 129.68, 129.59, 128.96, 123.67, 123.28, 122.48, 121.80, 119.68, 115.05, 41.87, 33.49, 33.06, 27.05, 26.61, 22.31.

[0086]

[0087] 8g: White solid, yield 63%, m.p. 118.3-121.2 °C; HRMS (ESI) m / z: calcd for C 25 H 23 N5OS[M+H] + :442.1709, found:442.1702. 1 H NMR (500 MHz, CDC13 ) δ 8.41 (d, J = 8.1 Hz, 1H), 8.30 (d, J = 8.0 Hz, 1H), 8.07 (s, 1H), 7.59 - 7.51 (m, 6H), 7.42 - 7.36 (m, 2H), 7.31 (t, J = 7.5 Hz, 1H), 4.44 (t, J = 6.7 Hz, 2H, -NCH2), 3.49 (t, J = 6.8 Hz, 2H, -SCH2), 2.57 (s, 3H), 2.08 - 2.03 (m, 2H), 1.99 (dd, J = 14.8, 7.4 Hz, 2H). 13C NMR (126 MHz, CDC13) δ 161.62, 154.42, 143.17, 137.17, 133.79, 133.69, 130.25, 129.93, 129.68, 129.59, 128.95, 123.98, 123.66, 123.25, 122.47, 121.80, 119.66, 115.06, 41.90, 33.04, 26.87, 26.71, 22.32.

[0088]

[0089] 8h: White solid, yield 56%, m.p. 131.3-133.6 °C; HRMS (ESI) m / z: calcd for C 27 H 27 N5OS[M+H] + : 470.2016, found: 470.2015. 1 H NMR (500 MHz, CDC13) δ 8.41 (d, J = 8.1 Hz, 1H), 8.30 (d, J = 7.8 Hz, 1H), 8.06 (s, 1H), 7.53 (t, J = 7.2 Hz, 1H), 7.45 (d, J = 8.2 Hz, 2H), 7.38 (dd, J = 17.3, 9.4 Hz, 4H), 7.31 (t, J = 7.3 Hz, 1H), 4.43 (s, 2H, -NCH2), 3.48 (t, J = 6.9 Hz, 2H, -SCH2), 2.74 (q, J = 7.4 Hz, 2H), 2.56 (s, 3H), 2.04 (d, J = 6.3 Hz, 2H), 2.00 - 1.93 (m, 2H), 1.29 (t, J = 7.6 Hz, 3H). 13 C NMR (126 MHz, CDC13) δ 161.62, 154.42, 143.17, 137.17, 133.79, 133.69, 130.25, 129.93, 129.68, 129.59, 128.95, 123.98, 123.66, 123.25, 122.47, 121.80, 119.66, 115.06, 41.90, 33.04, 26.87, 26.71, 22.32.

[0090]

[0091] 8i: White solid, yield 64%, m.p. 144.6-146.5 °C; HRMS (ESI) m / z: calcd for C 25 H22 ClN5OS[M+H] + : 476.1313, found: 476.1312. 1 H NMR (500 MHz, CDC13 ) δ 8.32 (s, 1H), 8.27 (d, J = 7.9 Hz, 1H), 8.17 (d, J = 8.3 Hz, 1H), 7.58 (d, J = 7.4 Hz, 1H), 7.55 - 7.49 (m, 5H), 7.38 (d, J = 8.4 Hz, 1H), 7.31 (t, J = 7.5 Hz, 1H), 4.46 (t, J = 6.9 Hz, 2H, -NCH2), 3.50 (t, J = 7.0 Hz, 2H, -SCH2), 2.52 (s, 3H), 2.06 (dd, J = 14.5, 6.6 Hz, 2H), 2.01 - 1.94 (m, 2H). 13 C NMR (126 MHz, CDC13 ) δ 161.68, 154.45, 138.35, 136.67, 136.31, 134.04, 132.25, 131.26, 130.19, 129.31, 128.70, 125.36, 125.21, 123.49, 122.57, 121.80, 119.85, 114.99, 41.91, 33.19, 26.82, 26.70, 21.51.

[0092]

[0093] 8j: White-like solid, yield 61%, m.p. 139.6-142.7 °C; HRMS (ESI) m / z: calcd for C 26 H 25 N5O2S[M+H] + : 472.1808, found: 472.1807. 1 H NMR (500 MHz, CDC13 ) δ 8.43 (d, J = 8.1 Hz, 1H), 8.31 (d, J = 7.9 Hz, 1H), 8.07 (s, 1H), 7.54 (t, J = 7.7 Hz, 1H), 7.44 (d, J = 8.9 Hz, 2H), 7.40 (t, J = 9.4 Hz, 2H), 7.32 (t, J = 7.6 Hz, 1H), 7.03 (d, J = 8.7 Hz, 2H), 4.45 (t, J = 6.9 Hz, 2H, -NCH2), 3.87 (s, 3H, -OCH3), 3.47 (t, J = 7.0 Hz, 2H, -SCH2), 2.57 (s, 3H), 2.05 (t, J = 7.1 Hz, 2H), 1.99 (dd, J = 15.0, 7.9 Hz, 2H).13 C NMR (126 MHz, CDC13) δ 161.76, 160.89, 154.58, 143.28, 137.11, 133.74, 129.72, 129.65, 129.02, 126.47, 125.73, 123.67, 123.18, 122.58, 121.81, 119.73, 115.15, 115.01, 114.58, 55.80, 55.77, 42.06, 32.99, 26.92, 26.73, 22.32.

[0094]

[0095] 8k: White solid, yield 65%, m.p. 175.2-176.6 °C; HRMS (ESI) m / z: calcd for C 21 H 23 N5O3S [M+H] + : 426.1064, found: 426.1600. 1 H NMR (500 MHz, CDC13) δ 8.17 (d, J = 8.0 Hz, 1H), 7.91 (s, 1H), 7.60 (s, 1H), 7.51 (t, J = 7.8 Hz, 1H), 7.38 (d, J = 8.4 Hz, 1H), 7.31 (t, J = 7.5 Hz, 1H), 4.46 (d, J = 6.9 Hz, 2H, -NCH2), 4.09 (s, 3H), 4.04 (s, 3H), 3.90 (s, 3H), 3.44 (t, J = 6.6 Hz, 2H, -SCH2), 2.05 - 1.96 (m, 4H). 13 C NMR (126 MHz, CDC13) δ 161.13, 154.34, 153.53, 150.01, 136.54, 128.95, 128.51, 123.13, 122.42, 119.54, 115.11, 109.07, 102.67, 56.33, 56.29, 46.05, 41.95, 33.49, 33.03, 27.05, 26.70, 8.75.

[0096]

[0097] 8l: White solid, yield 62%, m.p. 207.5-208.6 °C; HRMS (ESI) m / z: calcd for C 26 H 25 N5O3S [M+H] + : 488.1759, found: 488.1756.1 H NMR (500 MHz, CDC13 ) δ 8.18 (d, J = 7.9 Hz, 1H), 7.91 (s, 1H), 7.60 (s, 1H), 7.54 (ddd, J = 23.2, 10.5, 5.3 Hz, 6H), 7.39 (d, J = 8.4 Hz, 1H), 7.31 (t, J = 7.5 Hz, 1H), 4.46 (t, J = 7.0 Hz, 2H, -NCH2), 4.09 (s, 3H), 4.03 (s, 3H), 3.50 (t, J = 7.0 Hz, 2H, -SCH2), 2.05 (dd, J = 14.6, 7.6 Hz, 2H), 2.02 - 1.96 (m, 2H). 13 C NMR (126 MHz, CDC13 ) δ 161.13, 154.40, 153.53, 150.02, 136.56, 133.81, 130.24, 129.92, 128.94, 128.51, 123.98, 123.13, 122.41, 119.56, 115.12, 109.10, 102.68, 56.33, 56.29, 41.98, 33.06, 26.89, 26.83.

[0098]

[0099] 8m: White solid, yield 66%, m.p. 135.6-137.7 °C; HRMS (ESI) m / z: calcd for C 28 H 29 N5O2S [M+H] + : 516.2075, found: 516.2069. 1 H NMR (500 MHz, CDC13 ) δ 8.17 (d, J = 8.0 Hz, 1H), 7.91 (s, 1H), 7.60 (s, 1H), 7.51 (t, J = 7.7 Hz, 1H), 7.45 (d, J = 8.3 Hz, 2H), 7.38 (dd, J = 12.4, 8.5 Hz, 3H), 7.31 (t, J = 7.5 Hz, 1H), 4.45 (t, J = 6.8 Hz, 2H, -NCH2), 4.09 (s, 3H), 4.03 (s, 3H), 3.48 (t, J = 7.0 Hz, 2H, -SCH2), 2.74 (q, J = 7.6 Hz, 2H), 2.07 - 2.02 (m, 2H), 1.98 (dt, J = 13.9, 6.9 Hz, 2H), 1.28 (t, J = 7.6 Hz, 3H). 13C NMR (126 MHz, CDC13) δ 161.12, 154.38, 153.53, 150.02, 146.83, 136.57, 131.40, 129.29, 128.95, 128.52, 123.96, 123.13, 122.40, 119.55, 115.13, 109.11, 102.68, 56.33, 56.29, 42.01, 33.02, 28.74, 26.91, 26.83, 15.44.

[0100]

[0101] 8n: White solid, yield 68%, m.p. 153.4-156.6 °C; HRMS (ESI) m / z: calcd for C 26 H 24 ClN5O3S [M+H] + : 522.1375, found: 522.1367. 1 H NMR (500 MHz, CDC13) δ 8.19 (d, J = 7.9 Hz, 1H), 7.94 (s, 1H), 7.61 (s, 1H), 7.53 (s, 5H), 7.40 (d, J = 8.4 Hz, 1H), 7.33 (t, J = 7.6 Hz, 1H), 4.49 (t, J = 6.8 Hz, 2H, -NCH2), 4.10 (s, 3H), 4.05 (s, 3H), 3.51 (t, J = 7.0 Hz, 2H, -SCH2), 2.08 - 2.04 (m, 2H), 2.01 (d, J = 7.1 Hz, 2H). 13 C NMR (126 MHz, CDC13) δ 161.28, 154.47, 153.66, 150.11, 136.43, 136.32, 132.25, 130.19, 129.60, 128.99, 128.61, 125.21, 124.86, 123.18, 122.58, 119.63, 119.43, 115.19, 109.12, 102.69, 56.42, 56.31, 42.11, 33.21, 26.87, 26.84.

[0102]

[0103] 8o: White solid, yield 63%, m.p. 171.3-173.8 °C; HRMS (ESI) m / z: calcd for C 27 H 27 N5O4S [M+H] +: 518.1862, found: 518.1862. 1 H NMR (500 MHz, CDC13) δ 8.18 (d, J = 7.8 Hz, 1H), 7.93 (s, 1H), 7.61 (s, 1H), 7.51 (t, J = 7.8 Hz, 1H), 7.44 (d, J = 8.8 Hz, 2H), 7.40 (d, J = 8.4 Hz, 1H), 7.32 (t, J = 7.5 Hz, 1H), 7.03 (d, J = 8.8 Hz, 2H), 4.47 (d, J = 7.1 Hz, 2H, -NCH2), 4.10 (s, 3H), 4.04 (s, 3H), 3.87 (s, 3H), 3.47 (t, J = 7.0 Hz, 2H, -SCH2), 2.07 - 2.02 (m, 2H), 2.02 - 1.96 (m, 2H). 13 CNMR (126 MHz, CDC13) δ 161.07, 160.76, 154.43, 153.45, 149.93, 136.38, 128.85, 128.44, 126.34, 125.59, 123.02, 122.36, 119.46, 115.06, 114.88, 114.52, 108.99, 102.56, 56.26, 56.18, 55.68, 41.97, 32.86, 26.81, 26.71.

[0104]

[0105] 8p: white needle crystal, yield 65%, m.p. 168.2-169.8 °C; HRMS (ESI) m / z: calcd for C 19 H 17 Cl2N5OS[M+H] + : 434.0609, found: 434.0609. 1 H NMR (500 MHz, CDC13) δ 8.58 (s, 1H), 8.33 (s, 1H), 8.17 (d, J = 7.4 Hz, 1H), 7.61 - 7.57 (m, 1H), 7.39 (d, J = 8.5 Hz, 1H), 7.34 (t, J = 7.6 Hz, 1H), 4.41 (t, J = 7.1 Hz, 2H, -NCH2), 3.91 (s, 3H), 3.44 (t, J = 7.0 Hz, 2H, -SCH2), 2.04 - 1.96 (m, 4H). 13CNMR (126 MHz, CDC13) δ 160.00, 154.29, 144.72, 137.74, 137.32, 133.26, 132.70, 130.73, 125.00, 123.98, 123.89, 123.09, 117.96, 115.36, 42.22, 33.51, 32.90, 27.04, 26.48.

[0106]

[0107] 8q: white solid, yield 63%, m.p. 151.1-153.6 °C; HRMS (ESI) m / z: calcd for C 24 H 19 Cl2N5OS [M+H] + : 496.0767, found: 496.0766. 1 H NMR (500 MHz, CDC13) δ 8.57 (s, 1H), 8.33 (s, 1H), 8.17 (d, J = 7.4 Hz, 1H), 7.59 (d, J = 7.3 Hz, 1H), 7.57-7.56 (m, 4H), 7.55-7.53 (m, 1H), 7.39 (d, J = 8.5 Hz, 1H), 7.34 (t, J = 7.5 Hz, 1H), 4.42 (t, J = 7.3 Hz, 2H, -NCH2), 3.49 (t, J = 7.1 Hz, 2H, -SCH2), 2.07-2.03 (m, 2H), 1.99-1.95 (m, 2H). 13 C NMR (126 MHz, CDC13) δ 159.99, 154.33, 137.73, 137.32, 133.77, 133.25, 132.69, 130.79, 130.73, 130.29, 129.95, 125.00, 123.96, 123.88, 123.08, 117.96, 115.36, 46.09, 42.24, 32.93, 26.86, 26.59, 8.76.

[0108]

[0109] 8r: white needle crystal, yield 65%, m.p. 150.3-152.3 °C; HRMS (ESI) m / z: calcd for C 26 H 23 Cl2N5OS [M+H] + : 524.1086, found: 524.1079. 1H NMR (500 MHz, CDC13) δ 8.58 (s, 1H), 8.33 (s, 1H), 8.17 (d, J = 7.5 Hz, 1H), 7.58 (dd, J = 11.5, 4.2 Hz, 1H), 7.45 (d, J = 8.4 Hz, 2H), 7.40 - 7.34 (m, 4H), 4.41 (t, J = 7.2 Hz, 2H, -NCH2), 3.47 (t, J = 7.1 Hz, 2H, -SCH2), 2.74 (q, J = 7.6 Hz, 2H), 2.07 - 2.02 (m, 2H), 1.99 - 1.94 (m, 2H), 1.29 (t, J = 7.6 Hz, 3H). 13 C NMR (126 MHz, CDC13) δ 159.98, 154.30, 146.88, 137.72, 137.35, 133.26, 132.69, 131.38, 130.80, 130.74, 129.31, 125.02, 123.95, 123.88, 123.06, 117.96, 115.37, 42.26, 32.90, 28.74, 26.89, 26.59, 15.43.

[0110]

[0111] 8s: white solid, yield 70%, m.p. 197.3-200.6 °C; HRMS (ESI) m / z: calcd for C 24 H 18 Cl3N5OS [M+H] + : 530.0377, found: 530.0376. 1 H NMR (500 MHz, CDC13) δ 8.58 (s, 1H), 8.34 (s, 1H), 8.17 (d, J = 7.5 Hz, 1H), 7.59 (t, J = 7.3 Hz, 1H), 7.53 (s, 4H), 7.39 (d, J = 8.5 Hz, 1H), 7.34 (t, J = 7.6 Hz, 1H), 4.42 (t, J = 7.2 Hz, 2H, -NCH2), 3.50 (t, J = 7.1 Hz, 2H, -SCH2), 2.07 - 2.03 (m, 2H), 1.99 - 1.94 (m, 2H). 13C NMR (126 MHz, CDC13) δ 160.00, 154.38, 137.76, 137.31, 136.36, 133.24, 132.72, 132.21, 130.79, 130.73, 130.22, 125.19, 124.98, 123.97, 123.91, 123.09, 117.97, 115.33, 42.19, 33.05, 26.84, 26.58.

[0112]

[0113] 8t: white solid, yield 68%, m.p. 163.4-166.3 °C; HRMS (ESI) m / z: calcd for C 25 H 21 Cl2N5O2S [M+H] + : 526.0877, found: 526.0871. 1 H NMR (500 MHz, CDC13) δ 8.58 (s, 1H), 8.33 (s, 1H), 8.17 (d, J = 7.9 Hz, 1H), 7.58 (t, J = 7.7 Hz, 1H), 7.46 - 7.42 (m, 2H), 7.39 (d, J = 8.4 Hz, 1H), 7.34 (t, J = 7.5 Hz, 1H), 7.03 (d, J = 8.9 Hz, 2H), 4.41 (s, 2H, -NCH2), 3.88 (s, 3H, -OCH3), 3.46 (t, J = 6.9 Hz, 2H, -SCH2), 2.00 (dd, J = 31.0, 6.6 Hz, 4H). 13 C NMR (126 MHz, CDC13) δ 160.91, 159.98, 154.47, 137.72, 137.34, 133.26, 132.69, 130.80, 130.73, 126.42, 125.70, 125.01, 123.97, 123.88, 123.07, 117.96, 115.37, 115.02, 55.82, 42.25, 32.85, 26.90, 26.59.

[0114]

[0115] 9a: yellow solid, yield 62%, m.p. 103.1-105.6 °C; HRMS (ESI) m / z: calcd for C 22 H 25 N5OS [M+H] + : 408.1862, found: 408.1858.1 H NMR (500 MHz, CDC13 ) δ 8.34 (s, 1H), 8.27 (d, J = 8.0 Hz, 1H), 8.17 (d, J = 8.3 Hz, 1H), 7.58 (d, J = 8.2 Hz, 1H), 7.53 (t, J = 7.7 Hz, 1H), 7.39 (d, J = 8.4 Hz, 1H), 7.30 (t, J = 7.5 Hz, 1H), 4.44 - 4.35 (m, 2H, -NCH2), 3.90 (s, 3H, -NCH3), 3.35 (t, J = 7.3 Hz, 2H, -SCH2), 2.52 (s, 3H), 1.87 - 1.79 (m, 4H), 1.58 - 1.51 (m, 4H). 13 C NMR (126 MHz, CDC13 ) δ 161.69, 154.55, 138.31, 136.74, 133.99, 131.27, 129.26, 128.71, 125.40, 123.40, 122.50, 121.76, 119.85, 115.16, 42.66, 33.47, 33.33, 29.33, 28.41, 27.44, 26.57, 21.51.

[0116]

[0117] 9b: White solid, yield 59%, m.p. 113.3-115.2 °C; HRMS (ESI) m / z: calcd for C 27 H 27 N5OS [M + H] + : 470.2012, found: 470.2015. 1 H NMR (500 MHz, CDC13 ) δ 8.34 (s, 1H), 8.27 (d, J = 8.0 Hz, 1H), 8.17 (d, J = 8.3 Hz, 1H), 7.58 (d, J = 8.2 Hz, 1H), 7.53 (t, J = 7.7 Hz, 1H), 7.39 (d, J = 8.4 Hz, 1H), 7.30 (t, J = 7.5 Hz, 1H), 4.44 - 4.35 (m, 2H, -NCH2), 3.90 (s, 3H, -NCH3), 3.35 (t, J = 7.3 Hz, 2H, -SCH2), 2.52 (s, 3H), 1.87 - 1.79 (m, 4H), 1.58 - 1.51 (m, 4H). 13CNMR (126 MHz, CDCI3) δ 161.58, 154.58, 138.24, 136.83, 133.91, 131.25, 130.20, 129.91, 129.23, 128.69, 125.49, 124.00, 123.39, 122.41, 121.75, 119.81, 115.11, 42.60, 33.35, 29.15, 28.50, 27.44, 26.61, 21.51.

[0118]

[0119] 9c: White solid, yield 58%, m.p. 111.1-113.4 °C; HRMS (ESI) m / z: calcd for C 29 H 31 N5OS[M+H] + : 498.2330, found: 498.2328. 1 H NMR (500 MHz, CDCI3) δ 8.33 (s, 1 H), 8.27 (d, J = 8.0 Hz, 1 H), 8.17 (d, J = 8.3 Hz, 1 H), 7.57 (d, J = 8.3 Hz, 1 H), 7.52 (t, J = 7.7 Hz, 1 H), 7.46 (d, J = 8.2 Hz, 2 H), 7.38 (t, J = 6.8 Hz, 3 H), 7.30 (t, J = 7.6 Hz, 1 H), 4.44 - 4.34 (m, 2 H), 3.39 (t, J = 7.3 Hz, 2 H), 2.74 (q, J = 7.6 Hz, 2 H), 2.51 (d, J = 8.7 Hz, 3 H), 1.89 - 1.84 (m, 2 H), 1.84 - 1.78 (m, 2 H), 1.58 - 1.52 (m, 4 H), 1.28 (t, J = 7.6 Hz, 3 H). 13 CNMR (126 MHz, CDCI3) δ 161.58, 154.55, 146.78, 138.24, 136.81, 133.91, 131.43, 131.24, 129.28, 129.24, 128.68, 125.47, 123.96, 123.39, 122.41, 121.75, 119.80, 115.12, 42.61, 33.28, 29.15, 28.74, 28.51, 27.44, 26.62, 21.52, 15.45.

[0120]

[0121] 9d: White solid, yield 61%, m.p. 99.6-112.3 °C; HRMS (ESI) m / z: calcd for C 27 H 26 ClN5OS[M+H] + :504.1629, found:504.1625. 1 H NMR (500 MHz, CDC13) δ 8.33 (s, 1H), 8.27 (d, J = 8.0 Hz, 1H), 8.17 (d, J = 8.3 Hz, 1H), 7.57 (d, J = 8.3 Hz, 1H), 7.54 (s, 4H), 7.51 (d, J = 8.1 Hz, 1H), 7.38 (d, J = 8.4 Hz, 1H), 7.30 (t, J = 7.5 Hz, 1H), 4.43 - 4.35 (m, 2H, -NCH2), 3.41 (t, J = 7.3 Hz, 2H, -SCH2), 2.52 (s, 3H), 1.86 (d, J = 6.9 Hz, 2H), 1.82 (d, J = 6.7 Hz, 2H), 1.55 (d, J = 3.3 Hz, 4H). 13 C NMR (126 MHz, CDC13) δ 161.59, 154.63, 138.26, 136.83, 136.28, 133.93, 132.31, 131.25, 130.18, 129.23, 128.69, 125.49, 125.22, 123.41, 122.42, 121.76, 119.83, 115.10, 42.58, 33.48, 29.11, 28.48, 27.44, 26.59, 21.52.

[0122]

[0123] 9e: White solid, yield 68%, m.p. 99.2-101.6 °C; HRMS (ESI) m / z: calcd for C 28 H 29 N5O2S[M+H] + :500.2119, found:500.2120. 1H NMR (500 MHz, CDC13) δ 8.33 (s, 1H), 8.27 (d, J = 8.0 Hz, 1H), 8.17 (d, J = 8.3 Hz, 1H), 7.57 (d, J = 8.2 Hz, 1H), 7.52 (t, J = 7.8 Hz, 1H), 7.45 (d, J = 8.8 Hz, 2H), 7.38 (d, J = 8.5 Hz, 1H), 7.30 (t, J = 7.6 Hz, 1H), 7.04 (d, J = 8.8 Hz, 2H), 4.42 - 4.36 (m, 2H, -NCH2), 3.87 (s, 3H, -OCH3), 3.38 (t, J = 7.3 Hz, 2H, -SCH2), 2.52 (s, 3H), 1.87 - 1.83 (m, 2H), 1.83 - 1.78 (m, 2H), 1.57 - 1.52 (m, 4H). 13 C NMR (126 MHz, CDC13) δ 161.58, 160.86, 154.72, 138.24, 136.83, 133.91, 131.25, 129.24, 128.69, 126.52, 125.72, 125.49, 123.39, 122.41, 121.75, 119.81, 115.11, 114.98, 55.80, 42.61, 33.25, 29.17, 28.49, 27.44, 26.61, 21.51.

[0124]

[0125] 9f: White solid, yield 65%, m.p. 112.2-114.1 °C; HRMS (ESI) m / z: calcd for C 22 H 25 N5OS [M + H] + : 408.1854, found: 408.1858. 1 H NMR (500 MHz, CDC13) δ 8.42 (d, J = 8.1 Hz, 1H), 8.30 (d, J = 8.0 Hz, 1H), 8.06 (s, 1H), 7.54 (t, J = 7.8 Hz, 1H), 7.39 (t, J = 9.4 Hz, 2H), 7.30 (t, J = 7.6 Hz, 1H), 4.43 - 4.32 (m, 2H, -NCH2), 3.90 (s, 3H, -NCH3), 3.35 (t, J = 7.3 Hz, 2H, -SCH2), 2.56 (s, 3H), 1.88 - 1.79 (m, 4H), 1.59 - 1.51 (m, 4H). 13C NMR (126 MHz, CDC13) δ 161.59, 154.55, 143.06, 137.29, 133.68, 129.61, 129.55, 128.95, 123.59, 123.34, 122.37, 121.76, 119.65, 115.17, 42.54, 33.47, 33.31, 29.34, 28.41, 27.43, 26.59, 22.32.

[0126]

[0127] 9g: White solid, yield 66%, m.p. 101.4-103.6 °C; HRMS (ESI) m / z: calcd for C 27 H 27 N5OS [M+H] + : 470.2010, found: 470.2015. 1 H NMR (500 MHz, CDC13) δ 8.42 (d, J = 8.1 Hz, 1H), 8.30 (d, J = 7.9 Hz, 1H), 8.06 (s, 1H), 7.57 (q, J = 5.7 Hz, 4H), 7.53 (dd, J = 9.0, 3.3 Hz, 2H), 7.39 (t, J = 9.0 Hz, 2H), 7.30 (t, J = 7.6 Hz, 1H), 4.43 - 4.33 (m, 2H, -NCH2), 3.40 (t, J = 7.3 Hz, 2H, -SCH2), 2.56 (s, 3H), 1.90 - 1.84 (m, 2H), 1.84 - 1.78 (m, 2H), 1.59 - 1.51 (m, 4H). 13 C NMR (126 MHz, CDC13) δ 161.55, 154.58, 143.02, 137.35, 133.87, 133.69, 130.20, 129.91, 129.59, 129.53, 128.95, 124.00, 123.59, 123.40, 122.32, 121.75, 119.65, 115.15, 42.54, 33.35, 29.16, 28.50, 27.44, 26.62, 22.30.

[0128]

[0129] 9h: White solid, yield 57%, m.p. 117.2-119.8 °C; HRMS (ESI) m / z: calcd for C 29 H 31 N5OS [M+H] +:498.2333, found:498.2328. 1 H NMR (500 MHz, CDC13 ) δ 8.42 (d, J = 8.1 Hz, 1H), 8.30 (d, J = 8.0 Hz, 1H), 8.06 (s, 1H), 7.54 (t, J = 7.8 Hz, 1H), 7.46 (d, J = 8.1 Hz, 2H), 7.40 - 7.35 (m, 4H), 7.30 (t, J = 7.5 Hz, 1H), 4.42 - 4.33 (m, 2H, -NCH2), 3.39 (t, J = 7.2 Hz, 2H, -SCH2), 2.76 - 2.71 (m, 2H), 2.56 (s, 3H), 1.85 (d, J = 7.1 Hz, 2H), 1.82 (d, J = 7.4 Hz, 2H), 1.55 (s, 4H), 1.28 (t, J = 7.6 Hz, 3H). 13 C NMR (126 MHz, CDC13 ) δ 161.58, 154.56, 146.78, 143.05, 133.70, 129.61, 129.54, 129.27, 128.97, 123.97, 123.59, 122.35, 121.76, 119.66, 115.17, 42.58, 33.30, 29.17, 28.74, 28.50, 27.45, 26.63, 22.31, 15.42.

[0130]

[0131] 9i: White solid, yield 64%, m.p. 136.2-138.1 °C; HRMS (ESI) m / z: calcd for C 27 H 26 ClN5OS[M+H] + :504.1626, found:504.1625. 1 H NMR (500 MHz, CDC13 ) δ 8.42 (d, J = 8.1 Hz, 1H), 8.30 (d, J = 8.0 Hz, 1H), 8.06 (s, 1H), 7.54 (t, J = 7.8 Hz, 1H), 7.46 (d, J = 8.1 Hz, 2H), 7.40 - 7.35 (m, 4H), 7.30 (t, J = 7.5 Hz, 1H), 4.42 - 4.33 (m, 2H, -NCH2), 3.39 (t, J = 7.2 Hz, 2H, -SCH2), 2.76 - 2.71 (m, 2H), 2.56 (s, 3H), 1.85 (d, J = 7.1 Hz, 2H), 1.82 (d, J = 7.4 Hz, 2H), 1.55 (s, 4H), 1.28 (t, J = 7.6 Hz, 3H). 13C NMR (126 MHz, CDC13) δ 161.67, 154.65, 143.14, 137.29, 136.29, 133.74, 130.19, 129.61, 129.01, 125.23, 123.62, 122.43, 121.78, 119.72, 115.20, 42.62, 33.50, 29.13, 28.47, 27.46, 26.60, 22.31.

[0132]

[0133] 9j: White solid, yield 62%, m.p. 119.4-121.6 °C; HRMS (ESI) m / z: calcd for C 28 H 29 N5O2S [M+H]+: 500.2119, found: 500.2120. 1 H NMR (500 MHz, CDC13) δ 8.42 (d, J = 8.1 Hz, 1H), 8.30 (d, J = 8.0 Hz, 1H), 8.06 (s, 1H), 7.54 (t, J = 7.8 Hz, 1H), 7.45 (d, J = 8.8 Hz, 2H), 7.39 (t, J = 9.1 Hz, 2H), 7.30 (t, J = 7.5 Hz, 1H), 7.04 (d, J = 8.8 Hz, 2H), 4.41 - 4.33 (m, 2H, -NCH2), 3.87 (s, 3H, -OCH3), 3.38 (t, J = 7.2 Hz, 2H, -SCH2), 2.56 (s, 3H), 1.88 - 1.84 (m, 2H), 1.82 (d, J = 7.6 Hz, 2H), 1.55 (s, 4H). 13 C NMR (126 MHz, CDC13) δ 161.56, 160.86, 154.72, 143.03, 137.34, 133.69, 129.60, 129.53, 128.96, 126.54, 125.73, 123.59, 123.39, 122.34, 121.76, 119.65, 115.16, 114.99, 55.80, 42.56, 33.26, 29.18, 28.49, 27.45, 26.63, 22.31.

[0134]

[0135] 9k: White solid, yield 60%, m.p. 158.3-160.7 °C; HRMS (ESI) m / z: calcd for C 23 H 27N5O3S [M+H] + Found: 454.1913. 1 H NMR (500 MHz, CDC13) δ 8.18 (d, J = 7.9 Hz, 1H), 7.93 (s, 1H), 7.61 (s, 1H), 7.52 (t, J = 7.7 Hz, 1H), 7.39 (d, J = 8.4 Hz, 1H), 7.31 (t, J = 7.5 Hz, 1H), 4.46 - 4.34 (m, 2H, -NCH2), 4.09 (s, 3H), 4.04 (s, 3H), 3.90 (s, 3H), 3.35 (t, J = 7.3 Hz, 2H, -SCH2), 1.84 (dd, J = 14.3, 7.0 Hz, 4H), 1.58 - 1.52 (m, 4H). 13 C NMR (126 MHz, CDC13) δ 161.07, 154.53, 153.47, 150.01, 136.71, 128.86, 128.49, 123.06, 122.28, 119.69, 119.53, 115.21, 109.12, 102.68, 56.32, 56.28, 42.62, 33.45, 33.33, 29.36, 28.42, 27.54, 26.60.

[0136]

[0137] 9l: White solid, yield 63%, m.p. 189.5-190.8 °C; HRMS (ESI) m / z: calcd for C 28 H 29 N5O3S [M+H] + Found: 516.2069. 1 H NMR (500 MHz, CDC13) δ 8.18 (d, J = 7.9 Hz, 1H), 7.93 (s, 1H), 7.60 (s, 1H), 7.59 - 7.49 (m, 6H), 7.39 (d, J = 8.4 Hz, 1H), 7.31 (t, J = 7.5 Hz, 1H), 4.44 - 4.35 (m, 2H, -NCH2), 4.09 (s, 3H), 4.03 (s, 3H), 3.40 (t, J = 7.3 Hz, 2H, -SCH2), 1.90 - 1.85 (m, 2H), 1.85 - 1.79 (m, 2H), 1.56 (s, 4H). 13C NMR (126 MHz, CDC13) δ 161.06, 154.58, 153.43, 149.96, 136.69, 133.84, 130.22, 129.91, 128.87, 128.47, 123.99, 123.06, 122.28, 119.67, 119.51, 115.22, 109.08, 102.63, 56.32, 56.28, 42.63, 33.33, 29.14, 28.50, 27.53, 26.62.

[0138]

[0139] 9m: white solid, yield 60%, m.p. 115.6-117.8 °C; HRMS (ESI) m / z: calcd for C 30 H 33 N5O3S[M+H] + : 544.2378, found: 544.2382. 1 H NMR (500 MHz, CDC13) δ 8.18 (d, J = 8.0 Hz, 1H), 7.95 (s, 1H), 7.61 (s, 1H), 7.52 (t, J = 7.7 Hz, 1H), 7.46 (d, J = 8.2 Hz, 2H), 7.40 (d, J = 8.5 Hz, 1H), 7.37 (d, J = 8.1 Hz, 2H), 7.31 (t, J = 7.5 Hz, 1H), 4.46 - 4.36 (m, 2H, -NCH2), 4.10 (s, 3H), 4.04 (s, 3H), 3.39 (t, J = 7.3 Hz, 2H, -SCH2), 2.74 (d, J = 7.6 Hz, 2H), 1.90 - 1.85 (m, 2H), 1.83 (d, J = 7.3 Hz, 2H), 1.56 (s, 3H), 1.28 (t, J = 7.6 Hz, 4H). 13 C NMR (126 MHz, CDC13) δ 161.06, 154.58, 153.43, 149.96, 136.69, 133.84, 130.22, 129.91, 128.87, 128.47, 123.99, 123.06, 122.28, 119.67, 119.51, 115.22, 109.08, 102.63, 56.32, 56.28, 42.63, 33.33, 29.14, 28.50, 27.53, 26.62.

[0140]

[0141] 9n: White solid, yield 62%, m.p. 177.3-179.2 °C; HRMS (ESI) m / z: calcd for C 28 H 28 ClN5O3S[M+H] + : 550.1689, found: 550.1680. 1 H NMR (500 MHz, CDC13) δ 8.18 (d, J = 7.9 Hz, 1H), 7.94 (s, 1H), 7.61 (s, 1H), 7.54 - 7.50 (m, 5H), 7.40 (d, J = 8.4 Hz, 1H), 7.31 (t, J = 7.5 Hz, 1H), 4.45 - 4.38 (m, 2H, -NCH2), 4.10 (s, 3H), 4.04 (s, 3H), 3.41 (t, J = 7.3 Hz, 2H, -SCH2), 1.90 - 1.86 (m, 2H), 1.85 - 1.80 (m, 2H), 1.56 (d, J = 3.3 Hz, 4H). 13 C NMR (126 MHz, CDC13) δ 161.11, 154.63, 153.48, 150.00, 149.01, 136.64, 136.27, 132.29, 130.18, 128.87, 128.50, 125.21, 123.08, 122.34, 119.62, 119.54, 115.23, 109.09, 102.64, 56.35, 56.28, 42.67, 33.46, 29.10, 28.47, 27.54, 26.60.

[0142]

[0143] 9o: White solid, yield 63%, m.p. 183.3-185.4 °C; HRMS (ESI) m / z: calcd for C 29 H 31 N5O4S[M+H] + : 546.2176, found: 546.2175. 1H NMR (500 MHz, CDC13) δ 8.17 (d, J = 7.9 Hz, 1H), 7.93 (s, 1H), 7.60 (s, 1H), 7.52 (t, J = 7.6 Hz, 1H), 7.45 (d, J = 8.9 Hz, 2H), 7.39 (d, J = 8.4 Hz, 1H), 7.31 (t, J = 7.5 Hz, 1H), 7.03 (d, J = 8.9 Hz, 2H), 4.43 - 4.36 (m, 2H, -NCH2), 4.09 (s, 3H), 4.03 (s, 3H), 3.87 (s, 3H), 3.38 (t, J = 7.3 Hz, 2H, -SCH2), 1.89 - 1.84 (m, 2H), 1.82 (d, J = 7.8 Hz, 2H), 1.55 (s, 4H). 13 C NMR (126 MHz, CDC13) δ 161.06, 160.87, 154.70, 153.47, 150.01, 136.73, 128.86, 128.49, 125.72, 123.06, 122.27, 119.53, 115.21, 114.99, 109.14, 102.69, 56.32, 56.28, 55.80, 42.64, 33.26, 29.18, 28.49, 27.55, 26.63.

[0144]

[0145] 9p: White needle crystal, yield 64%, m.p. 128.4-130.6 °C; HRMS (ESI) m / z: calcd for C 21 H 21 Cl2N5OS[M+H] + : 462.0926, found: 462.0922. 1 H NMR (500 MHz, CDC13) δ 8.58 (s, 1H), 8.33 (s, 1H), 8.17 (d, J = 7.4 Hz, 1H), 7.62 - 7.57 (m, 1H), 7.39 (d, J = 8.5 Hz, 1H), 7.33 (t, J = 7.6 Hz, 1H), 4.38 - 4.33 (m, 2H, -NCH2), 3.90 (s, 3H, -NCH3), 3.35 (t, J = 7.3 Hz, 2H, -SCH2), 1.87 - 1.83 (m, 2H), 1.83 - 1.78 (m, 2H), 1.57 - 1.52 (m, 4H). 13C NMR (126 MHz, CDC13) δ 159.91, 154.51, 137.60, 137.47, 133.25, 132.61, 130.72, 125.12, 123.93, 123.81, 122.94, 117.92, 115.45, 42.84, 33.47, 33.25, 29.38, 28.38, 27.33, 26.52.

[0146]

[0147] 9q: White solid, yield 62%, m.p. 169.3-171.2 °C; HRMS (ESI) m / z: calcd for C 26 H 23 Cl2N5OS [M+H] + : 524.1085, found: 524.1079. 1 H NMR (500 MHz, CDC13) δ 8.59 (s, 1H), 8.34 (s, 1H), 8.17 (d, J = 6.8 Hz, 1H), 7.57 (s, 6H), 7.39 (d, J = 7.8 Hz, 1H), 7.33 (s, 1H), 4.36 (s, 2H, -NCH2), 3.40 (s, 2H, -SCH2), 1.87 (s, 2H), 1.80 (s, 2H), 1.55 (s, 4H). 13 C NMR (126 MHz, CDC13) δ 159.92, 154.55, 137.60, 137.48, 133.85, 133.26, 132.62, 130.73, 130.24, 129.93, 125.13, 123.98, 123.94, 123.82, 122.94, 117.94, 115.46, 42.86, 33.28, 29.18, 28.47, 27.34, 26.56.

[0148]

[0149] 9r: White solid, yield 65%, m.p. 126.6-128.8 °C; HRMS (ESI) m / z: calcd for C 28 H 27 Cl2N5OS [M+H] + : 552.1393, found: 552.1392. 1H NMR (500 MHz, CDC13) δ 8.59 (s, 1H), 8.33 (s, 1H), 8.17 (d, J = 8.0 Hz, 1H), 7.59 (t, J = 7.8 Hz, 1H), 7.48 - 7.44 (m, 2H), 7.38 (t, J = 8.5 Hz, 3H), 7.33 (t, J = 7.6 Hz, 1H), 4.39 - 4.33 (m, 2H, -NCH2), 3.39 (t, J = 7.3 Hz, 2H, -SCH2), 2.74 (q, J = 7.6 Hz, 2H), 1.88 - 1.79 (m, 4H), 1.55 (s, 4H), 1.29 (t, J = 7.6 Hz, 3H). 13 C NMR (126 MHz, CDC13) δ 159.91, 154.52, 146.81, 137.60, 137.48, 133.26, 132.62, 131.44, 130.73, 129.29, 125.13, 123.95, 123.81, 122.94, 117.93, 115.46, 42.87, 33.23, 29.19, 28.74, 28.46, 27.34, 26.56, 15.44.

[0150]

[0151] 9s: white solid, yield 69%, m.p. 83.5-87.6 °C; HRMS (ESI) m / z: calcd for C 26 H 22 Cl3N5OS[M+H] + : 558.0690, found: 558.0689. 1 H NMR (500 MHz, CDC13) δ 8.58 (s, 1H), 8.33 (s, 1H), 8.17 (d, J = 8.1 Hz, 1H), 7.61 - 7.57 (m, 1H), 7.54 (s, 4H), 7.39 (d, J = 8.5 Hz, 1H), 7.33 (t, J = 7.6 Hz, 1H), 4.38 - 4.33 (m, 2H, -NCH2), 3.41 (t, J = 7.3 Hz, 2H, -SCH2), 1.89 - 1.85 (m, 2H), 1.83 - 1.78 (m, 2H), 1.57 - 1.53 (m, 4H). 13C NMR (126 MHz, CDCI3) δ 159.92, 154.60, 137.62, 137.47, 136.31, 133.25, 132.64, 132.29, 130.72, 130.20, 125.20, 125.12, 123.94, 123.83, 122.95, 117.95, 115.44, 42.84, 33.40, 29.14, 28.45, 27.34, 26.53.

[0152]

[0153] 9t: white solid, yield 62%, m.p. 156.6-157.8 °C; HRMS (ESI) m / z: calcd for C 27 H 25 Cl2N5O2S [M+H] + : 554.1185, found: 554.1184. 1 H NMR (500 MHz, CDCI3) δ 8.59 (s, 1 H), 8.33 (s, 1 H), 8.17 (d, J = 7.5 Hz, 1 H), 7.60 (dd, J = 1 1.5, 4.2 Hz, 1 H), 7.45 (d, J = 8.9 Hz, 2 H), 7.39 (d, J = 8.5 Hz, 1 H), 7.33 (t, J = 7.6 Hz, 1 H), 7.04 (d, J = 8.9 Hz, 2 H), 4.38 - 4.33 (m, 2 H, -NCH2), 3.88 (s, 3 H, -OCH3), 3.38 (t, J = 7.3 Hz, 2 H, -SCH2), 1.88 - 1.83 (m, 2 H), 1.83 - 1.77 (m, 2 H), 1.56 - 1.52 (m, 4 H). 13 C NMR (126 MHz, CDCI3) δ 160.88, 159.91, 154.68, 137.60, 137.48, 133.26, 132.62, 130.73, 126.51, 125.70, 125.13, 123.94, 123.81, 122.94, 117.93, 115.46, 115.00, 55.81, 42.87, 33.19, 29.20, 28.45, 27.34, 26.55.

[0154] <Example 3>

[0155] In vitro anti-tumor activity experiment

[0156] The inhibition rates of the tetrazole sulfide derivatives 8a-8t, 9a-9t on human hepatoma cells (Hep G2), human cervical cancer cells (HeLa), human lung cancer cells (H460) and human normal liver cells (LO2) at a concentration of 40 μmol / L were studied by CCK-8 method, with 5-fluorouracil and chlorinated two-faced needle alkali as positive control groups.

[0157] (1) Cell culture: The frozen cells were taken out, completely thawed in a 37°C water bath, alcohol disinfected, and then placed in a clean bench. Clean centrifuge tubes were taken, and the thawed cells and appropriate culture medium containing 10% fetal bovine serum were added and centrifuged. After resuspension into a cell suspension by adding appropriate 10% culture medium, the cells were transferred to a culture bottle for culture. After 12 hours, the cell state was observed.

[0158] (2) Plate seeding: when the cells grow densely on the bottle bottom (about 80%), the culture medium is removed, and the cells are washed with PBS buffer for 1-2 times. The cells are digested with 0.25% trypsin for 1-2 min, and then the digestion is terminated by adding culture medium and centrifuging. The supernatant is removed, and the cells are resuspended in culture medium. The cells are counted, and 100 μL of cells with a concentration of 50,000 / mL are inoculated in each well of a 96-well plate.

[0159] (3) Drug addition: the above-synthesized compounds 12a-12o, 13a-13o and positive control drugs (5-fluorouracil-5-Fu) and chlorinated two-faced needle alkali (NC) are dissolved in DMSO to prepare samples with a concentration of 40 μmol / L. Three replicate wells are set for each concentration, and 100 μL of each well. The blank control group is 100 μL of 1% DMEM medium per well. The cells are cultured for 24 hours.

[0160] (4) Test: the 96-well plate cultured for 24 hours is taken out, 10 μL of CCK-8 reagent is added to each well, and the plate is incubated in the dark for 2 hours. The absorbance is measured at 450 nm by a microplate reader. The inhibition rate formula is as follows:

[0161] Cell growth inhibition rate (%) = [(Ac-As)] / (Ac-Ab)]x100%

[0162] As: absorbance of experimental wells Ac: absorbance of control wells Ab: absorbance of blank wells

[0163] The IC50 values are shown in Table 1:

[0164] Table 1 Inhibition rates of tetrazole sulfide derivatives on selected cells at a concentration of 40 μmol / L

[0165]

[0166]

[0167] The diterpenoid-tetrazole sulfide derivatives provided by the application have certain anti-tumor effects, as shown in Table 1. Compound 8f has significant inhibitory effects on Hepg2, H460 and HeLa cells, and the inhibition rates are 63.29%±3.74%, 69.40%±1.80% and 51.37%±2.16% respectively at a concentration of 40 μmol / L. The compound is not only superior to the positive control drug 5-fluorouracil, but also equivalent to the parent structure chloro-diterpenoid, and has lower toxicity than chloro-diterpenoid. Most of the diterpenoid-tetrazole sulfide derivatives have less effect on normal cells LO2.

[0168] The number of devices and the scale of processing illustrated here are used to simplify the description of the application. Applications, modifications and variations of the application will be apparent to those skilled in the art.

[0169] Although the embodiments of the application have been disclosed as above, it is not limited to the applications and embodiments listed in the specification and examples, and can be fully applied to various fields suitable for the application. Those skilled in the art can easily make other modifications, and therefore the application is not limited to specific details and examples shown and described herein.

Claims

1. A dimeric tetrazole thioether derivative of norcoclaurine, characterized in that, The derivative has a structure as general formula or general formula ​ When R1 is hydrogen, R2 is methyl, R3 is any one of methyl, phenyl, ethylbenzene, chlorobenzene, methoxybenzene; When R1 is hydrogen, R2 is methyl, R3 is any one of methyl, phenyl, ethylbenzene, chlorobenzene, methoxybenzene; When R1, R2 are methoxy, R3 is any one of methyl, phenyl, ethylbenzene, chlorobenzene, methoxybenzene; When R1, R2 are chlorine, R3 is any one of methyl, phenyl, ethylbenzene, chlorobenzene, methoxybenzene.

2. A process for the preparation of tetrazole thioether derivatives of norcoclaurine, characterized by, The method comprises the following steps: Step one, acyl chloride compound 1 and methoxyamine hydrochloride are used as raw materials, compound 2 shown in formula 2 is synthesized under the action of potassium carbonate and in the solvent of ethyl acetate; Step two, compound 2, iodobenzene, palladium acetate and silver oxide are dissolved in acetic acid, compound 3 shown in formula 3 is synthesized by reaction under nitrogen protection; Step three, compound 3 is dissolved in anhydrous methanol, compound 4 shown in formula 4 is obtained by reaction under ultraviolet lamp irradiation; Step four, compound 4 and 1,4-dibromobutane or 1,6-dibromohexane are used as raw materials, compound 5 shown in formula 5 or compound 6 shown in formula 6 is synthesized under the action of sodium hydride and in the solvent of DMF; Step five, isothiocyanate compound and sodium azide are used as raw materials, compound 7 shown in formula 7 is synthesized under the action of zinc chloride and in the solvent of acetonitrile; Step six, compound 5 or compound 6 and compound 7 are used as raw materials, compound 8 shown in formula 8 or compound 9 shown in formula 9 is synthesized under the action of triethylamine and in the solvent of acetonitrile, which is a two-side needle alkali-tetrazole sulfide derivative derivative of general formula I or general formula II; When R1 is hydrogen, R2 is methyl, R3 is any one of methyl, phenyl, ethylbenzene, chlorobenzene, methoxybenzene; When R1 is hydrogen, R2 is methyl, R3 is any one of methyl, phenyl, ethylbenzene, chlorobenzene, methoxybenzene; When R1, R2 are methoxy, R3 is any one of methyl, phenyl, ethylbenzene, chlorobenzene, methoxybenzene; When R1, R2 are chlorine, R3 is any one of methyl, phenyl, ethylbenzene, chlorobenzene, methoxybenzene.

3. The method of claim 2, wherein the tetrazole thioether derivative of norclerodendrin is represented by the following formula: ###0002### 3 In step one, the molar ratio of methoxyamine hydrochloride, potassium carbonate and compound 1 is 1:2:

1.

4. The method of claim 2, wherein the tetrazole thioether derivative of norclerodendrin is represented by the following formula: ###0002### 4 In step two, the molar ratio of compound 2, iodobenzene, palladium acetate and silver oxide is 1:2:0.05:

2.

5. The method of claim 2, wherein the tetrazole thioether derivative of norclerodendrin is represented by the following formula: ###0002### 5 In step three, the ultraviolet lamp is selected as a 175 W ultraviolet high-pressure mercury lamp.

6. The method of claim 2, wherein the tetrazole thioether derivative of norclerodendrin is represented by the following formula: ###0006### 6 In step four, the molar ratio of compound 4, sodium hydride, 1,4-dibromobutane or 1,6-dibromohexane is 1:3:

5.

7. The method of claim 2, wherein the tetrazole thioether derivative of norclerodendrin is represented by the following formula: ###0003### 7 In step five, the molar ratio of sodium azide, zinc oxide and allyl isothiocyanate is 1.2:1.2:

1.

8. The method of claim 2, wherein the tetrazole thioether derivative of norclerodendrin is represented by the following formula: ###0005### 8 In step six, the molar ratio of compound 5 or compound 6, compound 7 and triethylamine is 1:1:1.5.

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