Hirsutine alkaloid derivatives, processes for their preparation and use

Abstract

The application provides a Hirsutine alkaloid derivative, a preparation method and application thereof, and belongs to the technical field of medicines.The Hirsutine alkaloid derivative has a structural formula shown in formula (I).The application also provides a specific preparation method of the Hirsutine alkaloid derivative.Experiments prove that part of the Hirsutine alkaloid derivatives involved in the application have strong inhibitory effect on influenza virus (H1N1), and can be used for preparing medicines for preventing or treating influenza virus infection.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of medicine, and particularly relates to a Hirsutine alkaloid derivative, a preparation method thereof and application thereof. BACKGROUND

[0002] Respiratory infections caused by influenza viruses, which usually manifest as seasonal epidemics and intermittent pandemics, not only seriously affect public health, but also bring heavy economic burden to the country and society. At present, the anti-influenza virus drugs approved for clinical use in China include two ion channel inhibitors: amantadine and rimantadine, and two neuraminidase inhibitors: oral oseltamivir and inhaled zanamivir. However, most influenza viruses have developed varying degrees of resistance to amantadine and oseltamivir. Recently, the FDA has approved baloxavir marboxil targeting viral polymerase. However, the clinical efficacy, drug resistance and cost are still of concern. Therefore, it is of great significance to develop and stockpile anti-influenza virus drugs with new mechanisms, new targets and new structures.

[0003] Hirsutine is a claviform monoterpene indole alkaloid isolated from Mitragyna hirsuta in 1965, which has strong central inhibition and vasodilation effects. In addition, Takayama et al. (1997) first described the inhibitory effect of Hirsutine and its synthetic derivatives on influenza A virus in vitro. They found that Hirsutine inhibited the replication of influenza A virus (H3N2) with an SI (CC 50 / EC 50 ) of 58.8. Kai Zhu's research group and Hiromitsu's research group designed and synthesized Hirsutine derivatives by retaining the tetracyclic parent structure and introducing different substituents into the side chain, respectively, to obtain compounds with good antihypertensive and antiviral activities. We retained the tetracyclic parent structure on the basis of the structure of Hirsutine and made structural modifications at the C-20 position to seek antiviral candidate compounds with stronger activity, lower toxicity and better drug properties. SUMMARY

[0004] The application provides a Hirsutine alkaloid derivative, a preparation method thereof and application thereof in the preparation of antiviral drugs, which can be used for preparing drugs for preventing or treating viral infections.

[0005] To solve the above technical problems, the application provides a compound with a structure shown in formula (I) and a pharmaceutically acceptable salt form:

[0006]

[0007] wherein R is selected from one of the following groups: wherein R5 is o-NHCO, m-NHCO, p-NHCO, o-NHCH2, m-NHCH2, p-NHCH2; or n = 1, 2, 3, 4, 5; or wherein n = 1, 2, 3, 4, 5, R6 is H or CH3; or or or NH. In the formula, R1 is substituted at any position on the benzene ring and is selected from one of the following groups: CH3, CH2CH3, F, Cl, Br, CN. In the formula, one of R2, R3, R4 is H, and the other two can be the same or different and are selected from H, CH3, OCH3, F, NO2, CF3, NH2, OH; or R4 is H, and R2 and R3 are one of the following groups forming a saturated or unsaturated five- or six-membered ring through a C, O, N, S atom:

[0008] The pharmaceutically acceptable salt refers to the salt of the above-mentioned compound and inorganic acid, organic acid, alkali metal or alkaline earth metal, etc. These salts include (but not limited to): (1) the salt with inorganic acid, such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, phosphoric acid; (2) the salt with organic acid, such as acetic acid, lactic acid, citric acid, succinic acid, fumaric acid, gluconic acid, benzoic acid, methane sulfonic acid, ethane sulfonic acid, benzene sulfonic acid, p-toluene sulfonic acid, oxalic acid, succinic acid, tartaric acid, maleic acid, or arginine (3) other salts, including the salt with alkali metal or alkaline earth metal (such as sodium, potassium, calcium or magnesium), ammonium salt or water-soluble amine salt (such as N-methylglucamine salt), lower alkanol ammonium salt and other pharmaceutically acceptable amine salt (such as methylamine salt, ethylamine salt, propylamine salt, dimethylamine salt, trimethylamine salt, diethylamine salt, triethylamine salt, t-butylamine salt, ethylenediamine salt, hydroxyethylamine salt, dihydroxyethylamine salt, trihydroxyethylamine salt, and amine salt formed by morpholine, piperazine, lysine, respectively), or other conventional "prodrugs".

[0009] The Hirsutine alkaloid derivative described in the present application can be prepared by the following method, which comprises the following steps:

[0010] First, the Hirsutine parent nucleus part of 1,4,6,7,12,12b-hexahydroindolo[2,3-a]quinolizine-3-carboxylic acid ethyl ester with or without substituent on the indole ring is hydrolyzed by 5 times of equivalent lithium hydroxide in a reaction solution of THF:H2O=1:2, and the pH is adjusted to about 3 by 2N hydrochloric acid to obtain 1,4,6,7,12,12b-hexahydroindolo[2,3-a]quinolizine-3-carboxylic acid hydrochloride with or without substituent on the indole ring; then the amide compound and the Hirsutine alkaloid derivative in the application with R being NH are formed by condensation with an amine compound with HATU as the condensing agent under the condition of a molar ratio of 1:1.5; and the amide compound is condensed with a substituted benzene compound connected with a carboxyl group, an acyl chloride group or a methylene bromide group under the condition of a molar ratio of 1:1.5 to obtain the Hirsutine alkaloid derivative in the application except that R is NH.

[0011] The required 1,4,6,7,12,12b-hexahydroindolo[2,3-a]quinolizine-3-carboxylic acid ethyl ester with or without substituent on the indole ring has a structural formula shown in formula (II), and can be prepared according to a known method.

[0012]

[0013] In the formula, R1 is substituted at any position of the benzene ring and is selected from one of the following groups: CH3, CH2CH3, F, Cl, Br, CN.

[0014] The application provides a Hirsutine alkaloid derivative with antiviral effect and a preparation method, an activity testing method and an application thereof. Experiments prove that the Hirsutine alkaloid derivative involved in the application has strong antiviral effect on influenza virus and has low cytotoxicity, meets the characteristics of high efficiency and low toxicity of antiviral drugs, and can be used for preparing antiviral drugs. BRIEF DESCRIPTION OF DRAWINGS

[0015] Figure 1 It is a reaction flow schematic diagram of Example 1 of the application;

[0016] Figure 2 It is a reaction flow schematic diagram of Example 2 of the application;

[0017] Figure 3 It is a reaction flow schematic diagram of Example 3 of the application;

[0018] Figure 4 It is a reaction flow schematic diagram of Example 4 of the application;

[0019] Fig. 5 is a structural schematic diagram of the Hirsutine alkaloid derivative in the application. DETAILED DESCRIPTION

[0020] Hirsutine widely exists in Uncaria rhynchophylla, which is used as a traditional Chinese medicine for treating convulsions and depression for a long time. Hirsutine, as a main indole alkaloid isolated from Uncaria rhynchophylla, belongs to a monoterpenoid indole alkaloid compound of koenine type, which is composed of four cyclic parent structures and two side chains. Hirsutine has been proved to have strong medicinal value and is widely used in the research of various biological activities. The present application modifies the partial parent ring of Hirsutine, aims to obtain the Hirsutine alkaloid derivative of the present application, and tests the activity of part of the compounds, and the obtained Hirsutine alkaloid derivative has good in vitro anti-influenza virus activity and low cytotoxicity, and can be applied to the field of antiviral drugs.

[0021] The present application will be further described in detail in combination with examples.

[0022] Example 1, preparation of 1,4,6,7,12,12b-hexahydroindolo[2,3-a]quinolizine-3-carboxylic acid ethyl ester with or without substituent on the indole ring

[0023] The present example aims to prepare the required Hirsutine partial parent core raw material, and the synthesis is based on the synthesis method reported by Reymundo A. Villa et al., and the preparation route is as shown in the attached Figure 1 figure.

[0024] The structure formula of 1,4,6,7,12,12b-hexahydroindolo[2,3-a]quinolizine-3-carboxylic acid ethyl ester with or without substituent on the indole ring described in the present example is as follows:

[0025]

[0026] R1 is substituted at any position on the benzene ring and is selected from one of the following groups: CH3, CH2CH3, F, Cl, Br, CN.

[0027] 1, preparation of 2-formyl-1H-indole-1-carboxylic acid tert-butyl ester

[0028] Into a 100 mL round bottom flask, weighed 1H-indole-2-carboxaldehyde (1.0 g, 6.89 mmol, 1.0 eq) and DMAP (84.18 mg, 0.69 mmol, 0.1 eq), dissolved in 10 mL THF, then added triethylamine (11.47 mL, 8.27 mmol, 1.2 eq), slowly added di-tert-butyl dicarbonate (18.04 g, 82.67 mmol, 1.2 eq) dissolved in 10 mL THF dropwise under ice bath, then transferred to room temperature 25 °C for 12 h. After TLC detection of the complete reaction, the solvent was evaporated under reduced pressure, extracted with dichloromethane for 3 times, combined the organic phase, washed once with saturated NaCl, dried over anhydrous magnesium sulfate, and finally column chromatography (PE:EA=80:1) to obtain 1.25 g of white solid, yield: 74%.

[0029] 2. Preparation of 2-(5-(ethoxycarbonyl)-1-((4-nitrophenyl)sulfonyl)-1,2,3,6- tetrahydropyridin-2-yl)-1H-indole-1-carboxylic acid tert-butyl ester

[0030] Into a 100 mL round bottom flask, weighed 1H-indole-2-carboxaldehyde (1.0 g, 6.89 mmol, 1.0 eq) and DMAP (84.18 mg, 0.69 mmol, 0.1 eq), dissolved in 10 mL THF, then added triethylamine (11.47 mL, 8.27 mmol, 1.2 eq), slowly added di-tert-butyl dicarbonate (18.04 g, 82.67 mmol, 1.2 eq) dissolved in 10 mL THF dropwise under ice bath, then transferred to room temperature 25 °C for 12 h. After TLC detection of the complete reaction, the solvent was evaporated under reduced pressure, extracted with dichloromethane for 3 times, combined the organic phase, washed once with saturated NaCl, dried over anhydrous magnesium sulfate, and finally column chromatography (PE:EA=80:1) to obtain 1.25 g of white solid, yield: 74%.

[0031] 3. Preparation of 6-(1H-indol-2-yl)-1-((4-nitrophenyl)sulfonyl)-1,2,5,6- tetrahydropyridine-3-carboxylic acid ethyl ester

[0032] Take 2-(5-(ethoxycarbonyl)-l-((4-nitrophenyl)sulfonyl)-l,2,3,6-tetrahydropyridin-2-yl)-lH-indole-l-carboxylic acid tert-butyl ester (1.97 g, 3.55 mmol, 1.0 eq) and 1.97 g SiO2 in a 100 mL round-bottom flask, add 20 mL toluene, heat under reflux conditions for 5 h, after TLC detection of the reaction is complete, evaporate the solvent under reduced pressure, dry, column chromatography (PE:EA = 2:1) to get yellow solid 1.31 g, yield: 81%.

[0033] 4, 6-(3-(2-hydroxyethyl)-lH-indol-2-yl)-l-((4-nitrophenyl)sulfonyl)-l,2,5,6-tetrahydropyridine-3-carboxylic acid ethyl ester preparation

[0034] Take 6-(lH-indol-2-yl)-l-((4-nitrophenyl)sulfonyl)-l,2,5,6-tetrahydropyridine-3-carboxylic acid ethyl ester (1.28 g, 2.81 mmol, 1.0 eq) in a 100 mL three-necked flask, N2 protection, add 30 mL anhydrous THF, ice bath slowly drop oxalyl chloride (0.71 mL, 8.43 mmol, 3.0 eq), after adding, transfer to room temperature 25 °C reaction 12 h, after TLC detection of the reaction is complete, ice bath slowly drop BH3-DMS (1.69 mL, 1.69 mmol, 6.0 eq), after adding, transfer to room temperature 25 °C reaction 4 h, after TLC detection of the reaction is complete, add saturated NaHCO3 solution to quench to no bubbles, evaporate THF under reduced pressure, extract with ethyl acetate three times, combine the organic phase, washed with saturated NaCl once, dry over anhydrous magnesium sulfate, finally column chromatography (PE:EA = 1:1) to get yellow solid 0.95 g, yield: 68%.

[0035] 5, 6-(3-(2-hydroxyethyl)-lH-indol-2-yl)-l,2,5,6-tetrahydropyridine-3-carboxylic acid ethyl ester preparation

[0036] Take 6-(3-(2-hydroxyethyl)-1H-indol-2-yl)-1-((4-nitrophenyl)sulfonyl)-1,2,5,6- tetrahydropyridine-3-carboxylic acid ethyl ester (950 mg, 1.90 mmol, 1.0 eq), p- toluene thiol (283 mg, 2.28 mmol, 1.2 eq) and anhydrous potassium carbonate (789 mg, 5.71 mmol, 3.0 eq) in a 100 mL round-bottom flask, add 20 mL acetonitrile to dissolve, react at 50 °C for 3 h, after TLC detection of the reaction completion, evaporate acetonitrile under reduced pressure, extract with dichloromethane for three times, combine the organic phase, wash once with saturated NaCl, dry over anhydrous magnesium sulfate, and purify by column chromatography (DCM:MeOH = 20:1) to obtain 0.45 g of white solid, yield: 75%.

[0037] 6, 1, 4, 6, 7, 12, 12b-hexahydroindolo[2, 3-a]quinoline-3-carboxylic acid ethyl ester (compound II-1)

[0038] Take triphenylphosphine (563 mg, 2.15 mmol, 1.5 eq) and imidazole (146 mg, 2.15 mmol, 1.5 eq) in a 100 mL three-necked flask, protect under N2, add 10 mL dichloromethane to dissolve, add iodine monomer (436 mg, 1.72 mmol, 2.0 eq) under ice bath, after 5 min, add 6-(3-(2-hydroxyethyl)-1H-indol-2-yl)-1,2,5,6-tetrahydropyridine-3-carboxylic acid ethyl ester (450 mg, 1.43 mmol, 1.0 eq), transfer to room temperature 25 °C for 1 h, after TLC detection of the reaction completion, add 2 mL triethylamine to the reaction solution, extract with dichloromethane and saturated NaHCO3 solution for three times, combine the organic phase, wash once with saturated NaCl, dry over anhydrous magnesium sulfate, and finally purify by column chromatography (PE:EA = 5:1) to obtain 300 mg of white solid, yield: 71%. 1 H NMR (400 MHz, CDCl3) 7.73 (br s, 1H), 7.42 (d, J = 7.7 Hz, 1H), 7.26 (d, J = 7.7 Hz, 1H), 7.16-7.08 (m, 2H), 7.03 (br s, 1H), 4.19 (q, J = 7.1 Hz, 2H), 3.79 (d, J = 16.5 Hz, 1H), 3.49 (d, J = 10.4 Hz, 1H), 3.28-3.17 (m, 2H), 3.04-2.95 (m, 1H), 2.82-2.67 (m, 3H), 2.49-2.41 (m, 1H), 1.29 (t, J = 7.1 Hz, 3H); 13CNMR (100 MHz, CDC13) 165.42, 136.15, 136.37, 133.64, 129.24, 126.62, 121.14, 119.56, 118.14, 110.62, 108.24, 60.35, 54.34, 53.23, 52.07, 31.21, 21.14, 14.35.

[0039]

[0040] 7. Preparation of 10-methyl-l,4,6,7,12,12-hexahydroindolo[2,3- a]quinoline-3-carboxylic acid ethyl ester (Compound II-2)

[0041] Prepared in the same manner as described in Example 1, steps 1-6, starting from 5-methyl-lH-indole-2-carboxaldehyde to give 3.61 g of white solid in 18% overall yield. 1 H NMR (400 MHz, DMSO-d6) δ 8.48 (s, 1H), 7.39 - 7.34 (m, 1H), 7.03 - 7.09 (d, J = 7.7 Hz, 1H), 6.97 - 6.91 (d, J = 5.1 Hz, 1H), 4.49 - 4.41 (m, 1H), 4.23 - 4.15 (q, J = 7.1 Hz, 2H), 3.90 - 3.85 (m, 1H), 3.77 - 3.68 (m, 1H), 3.28 - 3.19 (ddd, J = 12.1, 6.1, 4.2 Hz, 1H), 3.15 - 3.11 (m, 1H), 2.99 - 2.87 (m, 2H), 2.73 - 2.65 (m, 1H), 2.48 - 2.39 (m, 1H), 1.30 (t, J = 7.1 Hz, 3H). 13 C NMR (100 MHz, DMSO-d6) δ 167.23, 139.53, 135.99, 132.80, 130.19, 126.27, 125.84, 120.72, 119.59, 109.78, 107.51, 60.67, 55.26, 53.61, 52.15, 31.88, 21.48, 20.29, 14.21.

[0042]

[0043] 8. Preparation of 10-bromo-l,4,6,7,12,12-hexahydroindolo[2,3- a]quinoline-3-carboxylic acid ethyl ester (Compound II-3)

[0044] Prepared in the same manner as described in example 1, starting from 5-bromo-lH-indole-2-carboxaldehyde, to give 3.01 g of white solid in 17% overall yield. 1 H NMR (400 MHz, DMSO-d6) δ 9.07 (s, 1H), 7.56-7.49 (d, J = 2.7 Hz, 1H), 7.39-7.32 (dd, J = 7.3, 2.4 Hz, 1H), 7.21 (s, 1H), 6.97-6.89 (m, 1H), 4.49-4.41 (m, 1H), 4.23-4.15 (q, J = 7.1 Hz, 2H), 3.90-3.81 (m, 1H), 3.77-3.68 (m, 1H), 3.28-3.17 (m, 1H), 3.15-3.08 (m, 1H), 2.99-2.87 (m, 2H), 2.73-2.48 (m, 2H), 1.30 (t, J = 7.1 Hz, 3H). 13 CNMR (100 MHz, DMSO-d6) δ 167.22, 139.54, 136.15, 132.75, 128.07, 127.04, 122.65, 120.78, 112.45, 112.33, 107.50, 60.70, 55.30, 53.72, 51.98, 31.64, 21.58, 14.21.

[0045]

[0046] Example 2, Preparation of Hirsutine alkaloid derivatives modified with benzoic acid or heterocyclic benzoic acid containing different substituents

[0047] This reaction example uses compound II as starting material, hydrolysis is carried out under the condition of lithium hydroxide, then pH is adjusted to about 3 with 2N dilute hydrochloric acid, followed by condensation with mono-Boc protected aromatic diamine or aliphatic diamine, then de-Boc, the obtained intermediate is condensed with benzoic acid or heterocyclic benzoic acid containing different substituents to obtain the Hirsutine alkaloid derivatives modified with benzoic acid or heterocyclic benzoic acid containing different substituents, the preparation route is shown in the following figure. Figure 2

[0048] The Hirsutine alkaloid derivatives modified with benzoic acid or heterocyclic benzoic acid containing different substituents described in this example have the following structure:

[0049]

[0050] In the formula, R is selected from one of the following groups: wherein R5 is o-NHCO, m-NHCO, p-NHCO; or​ n = 1, 2, 3, 4, 5; or or NH. In the formula R1is substituted at any position on the benzene ring and is selected from one of the following groups: CH3, CH2CH3, F, Cl, Br, CN. In the formula one of R2, R3, R4is H and the other two can be the same or different and are selected from H, CH3, OCH3, F, NO2, CF3, NH2, OH; or R4is H and R2and R3are one of the following groups forming a saturated or unsaturated five or six membered ring through a C, O, N, S atom:

[0051] 1. Preparation of 1,4,6,7,12,12b-hexahydroindolo[2,3-a]quinolizine-3-carboxylic acid hydrochloride

[0052] In a 100 mL round bottom flask, 1,4,6,7,12,12b-hexahydroindolo[2,3-a]quinolizine-3-carboxylic acid (3.21 g, 10.8 mmol, 1.0 eq) was added, 11 mL THF and 22 mL water were added, LiOH-H2O (2.27 g, 54 mmol, 5.0 eq) was added, stirred at room temperature for 12 h, after TLC detection of the reaction was complete, 2N dilute hydrochloric acid was adjusted to pH about 3, suction filtration and washed with 10 mL water three times, and dried to give 1,4,6,7,12,12b-hexahydroindolo[2,3-a]quinolizine-3-carboxylic acid hydrochloride 2.51 g, yield 76%.

[0053] 2. Preparation of tert-butyl (2-(1,4,6,7,12,12b-hexahydroindolo[2,3-a]quinolin-3- formamido)phenyl)carbamate

[0054] In a 100 mL round bottom flask, 1,4,6,7,12,12b-hexahydroindolo[2,3-a]quinolizine-3-carboxylic acid (3.21 g, 10.8 mmol, 1.0 eq) was added, 11 mL THF and 22 mL water were added, LiOH-H2O (2.27 g, 54 mmol, 5.0 eq) was added, stirred at room temperature for 12 h, after TLC detection of the reaction was complete, 2N dilute hydrochloric acid was adjusted to pH about 3, suction filtration and washed with 10 mL water three times, and dried to give 1,4,6,7,12,12b-hexahydroindolo[2,3-a]quinolizine-3-carboxylic acid hydrochloride 2.51 g, yield 76%.

[0055] 3. Preparation of N-(2-aminophenyl)-l,4,6,7,12,12b-hexahydroindolo[2,3- a]quinoline-3-carboxamide

[0056] In a 100 mL single neck flask, tert-butyl (2-(l,4,6,7,12,12b-hexahydroindolo[2,3- a]quinoline-3-carboxamide)phenyl)carbamate (350 mg, 0.98 mmol, 1.0 eq) was added, 4 mL of trifluoroacetic acid was added, stirred at room temperature for 30 min. After TLC detection of complete, saturated NaHCO3 solution was added to the reaction solution to adjust the pH to about 8, saturated NaHCO3 solution and ethyl acetate were extracted three times, the organic phase was collected, dried over anhydrous magnesium sulfate, and column chromatography (DCM:MeOH=20:1) to obtain the target product 216 mg, yield 85%.

[0057] 4. Preparation of N-(2-(3-methoxy-4-methylbenzamido)phenyl)-l,4,6,7,12,12b- hexahydroindolo[2,3-a]quinoline-3-carboxamide (Compound I-1)

[0058] In a 100 mL single neck flask, 3-methoxy-4-methylbenzoic acid (111 mg, 0.67 mmol, 1.2 eq) and HATU (255 mg, 0.67 mmol, 1.2 eq) were added, followed by the addition of DMF 10 mL and DIPEA (292 μL, 1.68 mmol, 3.0 eq), after stirring at room temperature for 10 min, N-(2-aminophenyl)-l,4,6,7,12,12b-hexahydroindolo[2,3- a]quinoline-3-carboxamide (200 mg, 0.56 mmol, 1.0 eq) was finally added, stirred at room temperature overnight. After TLC detection of complete reaction, about 100 mL of water was added to the reaction solution, extracted with ethyl acetate three times, washed with saturated NaCl once, column chromatography (DCM:MeOH=25:1) to obtain white solid 180 mg, yield: 64%. 1H NMR (400 MHz, DMSO-d6) δ 10.79 (s, 1H), 9.92 (s, 1H), 9.58 (s, 1H), 7.67 - 7.60 (m, 1H), 7.57 - 7.53 (m, 1H), 7.49 - 7.42 (m, 2H), 7.36 (d, J = 7.7 Hz, 1H), 7.30 - 7.21 (m, 4H), 7.00 (ddd, J = 8.2, 7.0, 1.3 Hz, 1H), 6.93 (ddd, J = 8.0, 7.0, 1.1 Hz, 1H), 6.83 (br, 1H), 3.85 (s, 3H), 3.70 (d, J = 16.9 Hz, 1H), 3.46 - 3.41 (m, 1H), 3.17 - 3.08 (m, 2H), 2.92 - 2.84 (m, 1H), 2.81 - 2.73 (m, 1H), 2.67 - 2.61 (m, 1H), 2.54 (td, J = 11.2, 3.9 Hz, 1H), 2.19 (s, 4H). 13 CNMR (100 MHz, DMSO-d6) δ 165.66, 165.55, 157.80, 136.75, 135.48, 133.60, 133.04, 132.06, 131.71, 131.65, 130.97, 130.70, 126.99, 126.19, 126.11, 126.02, 125.96, 121.13, 120.09, 118.94, 118.18, 111.52, 109.56, 107.12, 55.91, 55.10, 53.73, 52.20, 31.67, 21.76, 16.64. HRMS (ESI) m / z: calcd for (C 31 H 31 O3N4+H) + ,507.2391; found: 507.2375.

[0059]

[0060] 5. Preparation of N-(2-(3-methylbenzamido)phenyl)-l,4,6,7,12,12b-hexahydroindolo[2,3- a]quinoline-3-carboxamide (Compound I-2)

[0061] Prepared in the same manner as described in Example 2, steps 1-4, except that one of the starting materials was 3-methylbenzoic acid, to give 173 mg of a white solid in 65% yield. 1H NMR (400 MHz, DMSO-c / 6) δ 10.85 (s, 1H), 10.00 (s, 1H), 9.60 (s, 1H), 7.82 - 7.74 (m, 2H), 7.68 - 7.63 (m, 1H), 7.62 - 7.57 (m, 1H), 7.47 - 7.42 (m, 2H), 7.40 (d, J = 7.8 Hz, 1H), 7.31 (d, J = 8.0 Hz, 1H), 7.29 - 7.25 (m, 2H), 7.05 (ddd, J = 8.2, 7.0, 1.3 Hz, 1H), 6.97 (td, J = 7.4, 1.1 Hz, 1H), 6.88 (br, 1H), 3.76 (d, J = 16.3 Hz, 1H), 3.51 (d, J = 10.7 Hz, 1H), 3.26 - 3.13 (m, 2H), 3.00 - 2.88 (m, 1H), 2.86 - 2.77 (m, 1H), 2.72 - 2.65 (m, 1H), 2.64 - 2.56 (m, 1H), 2.41 (s, 3H), 2.30 - 2.19 (m, 1H). 13 C NMR (100 MHz, DMSO-c / 6) δ 165.98, 165.37, 138.42, 136.70, 135.26, 134.64, 133.00, 132.89, 131.92, 131.64, 129.04, 128.52, 126.91, 126.20, 126.08, 126.02, 125.89, 125.17, 121.10, 118.89, 118.13, 111.47, 107.04, 55.05, 53.63, 52.14, 31.53, 21.64, 21.47. HRMS (ESI) m / z: calcd for (C 30 H 29 O2N4+H) + , 477.2285; found: 477.2272.

[0062]

[0063] 6. Preparation of N-(2-(3,4-dimethoxybenzamido)phenyl)-l,4,6,7,12,12b- hexahydroindolo[2,3-a]quinoline-3-carboxamide (Compound I-3)

[0064] Prepared in the same manner as in Example 2, steps 1-4, except that one of the starting materials was 3,4-dimethoxybenzoic acid, to give 219 mg of a white solid in 75% yield. 1H NMR (400 MHz, DMSO-d6) δ 10.84 (s, 1H), 9.93 (s, 1H), 9.61 (s, 1H), 7.67 - 7.54 (m, 4H), 7.39 (d, J = 7.7 Hz, 1H), 7.31 (d, J = 8.0 Hz, 1H), 7.29 - 7.23 (m, 2H), 7.12 (d, J = 8.5 Hz, 1H), 7.04 (ddd, J = 8.2, 7.0, 1.3 Hz, 1H), 6.96 (t, J = 6.9 Hz, 1H), 6.87 (br, 1H), 3.86 (s, 3H), 3.84 (s, 3H), 3.74 (d, J = 17.0 Hz, 1H), 3.47 (d, J = 13.0 Hz, 1H), 3.23 - 3.10 (m, 2H), 2.98 - 2.85 (m, 1H), 2.85 - 2.77 (m, 1H), 2.73 - 2.58 (m, 2H), 2.27 - 2.18 (m, 1H). 13 C NMR (100 MHz, DMSO-d6) δ 165.35, 165.30, 152.36, 148.87, 136.69, 135.18, 132.82, 131.94, 131.71, 131.58, 126.89, 126.62, 126.14, 126.02, 125.85, 121.48, 121.12, 118.91, 118.13, 111.62, 111.48, 111.13, 107.02, 56.17, 56.04, 55.05, 53.56, 52.12, 31.49, 21.59. HRMS (ESI) m / z: calcd for (C 31 H 31 O4N4+H) + ,523.2340; found: 523.2330.

[0065]

[0066] 7. Preparation of N-(2-(3-methoxybenzamido)phenyl)-l,4,6,7,12,12b- hexahydroindolo[2,3-a]quinoline-3-carboxamide (Compound I-4)

[0067] Prepared in the same manner as described in Example 2, steps 1-4, except that one of the starting materials was 3-methoxybenzoic acid, to give a white solid 157 mg, yield: 57%. 1H NMR (400 MHz, DMSO-c / 6) δ 10.85 (s, 1H), 10.02 (s, 1H), 9.61 (s, 1H), 7.69 - 7.64 (m, 1H), 7.56 (dt, J = 7.8, 1.3 Hz, 1H), 7.52 (t, J = 2.1 Hz, 1H), 7.48 (t, J = 7.9 Hz, 1H), 7.40 (d, J = 7.7 Hz, 1H), 7.31 (d, J = 8.0 Hz, 1H), 7.30 - 7.25 (m, 2H), 7.19 (ddd, J = 8.2, 2.6, 1.0 Hz, 1H), 7.05 (ddd, J = 8.1, 7.0, 1.3 Hz, 1H), 6.97 (td, J = 7.4, 1.1 Hz, 1H), 6.87 (br, 1H), 3.85 (s, 3H), 3.76 (d, J = 16.3 Hz, 1H), 3.50 (d, J = 10.7 Hz, 1H), 3.25 - 3.12 (m, 2H), 2.97 - 2.89 (m, 1H), 2.86 - 2.76 (m, 1H), 2.72 - 2.64 (m, 1H), 2.60 (td, J = 11.2, 3.9 Hz, 1H), 2.31 - 2.17 (m, 1H). 13 C NMR (100 MHz, DMSO-c / 6) δ 165.65, 165.53, 159.86, 136.77, 136.14, 135.39, 133.02, 131.97, 131.75, 131.61, 130.37, 126.99, 126.31, 126.14, 125.94, 121.16, 120.29, 118.96, 118.37, 118.19, 113.14, 111.54, 107.11, 55.88, 55.11, 53.70, 52.19, 31.61, 21.72. HRMS (ESI) m / z: calcd for (C 30 H 29 O3N4+H) + , 493.2234; found: 493.2221.

[0068]

[0069] 8. Preparation of N-(2-(3,4-dimethylbenzamido)phenyl)-l,4,6,7,12,12b- hexahydroindolo[2,3-a]quinoline-3-carboxamide (Compound 1-5)

[0070] Prepared in the same manner as described in Example 2, steps 1-4, except that one of the starting materials was 3,4-dimethylbenzoic acid, to give a white solid 201 mg, yield: 73%.1 H NMR (400 MHz, DMSO-c / 6) δ 10.81 (s, 1H), 9.90 (s, 1H), 9.57 (s, 1H), 7.72 (s, 1H), 7.67 (dd, J = 7.8, 2.0 Hz, 1H), 7.63 - 7.58 (m, 1H), 7.58 - 7.53 (m, 1H), 7.36 (d, J = 7.8 Hz, 1H), 7.28 (dd, J = 8.0, 3.2 Hz, 2H), 7.25 - 7.20 (m, 2H), 7.01 (ddd, J = 8.1, 7.0, 1.3 Hz, 1H), 6.93 (ddd, J = 8.0, 7.1, 1.1 Hz, 1H), 6.83 (br, 1H), 3.71 (d, J = 16.3 Hz, 1H), 3.49 - 3.40 (m, 1H), 3.21 - 3.08 (m, 2H), 2.94 - 2.85 (m, 1H), 2.82 - 2.72 (m, 1H), 2.68 - 2.61 (m, 1H), 2.55 (td, J = 11.3, 3.9 Hz, 1H), 2.28 (s, 3H), 2.26 (s, 3H), 2.24 - 2.13 (m, 1H). 13 C NMR (100 MHz, DMSO-c / 6) δ 165.86, 165.37, 141.30, 137.06, 136.69, 135.40, 132.98, 132.09, 131.96, 131.71, 131.57, 130.15, 128.99, 126.93, 126.13, 126.04, 125.92, 125.88, 125.52, 121.07, 118.87, 118.12, 111.46, 107.06, 55.04, 53.69, 52.15, 31.61, 21.70, 19.96, 19.91. HRMS (ESI) m / z: calcd for (C 31 H 31 O2N4+H) + , 491.2442; found: 491.2429.

[0071]

[0072] 9. Preparation of N-(2-benzoylamino phenyl)-l,4,6,7,12,12b-hexahydroindolo[2,3- a]quinoline-3-carboxamide (Compound 1-6)

[0073] Prepared in the same manner as in Example 2, steps 1-4, except that one of the starting materials was benzoic acid, to give 189 mg of a white solid in 73% yield. 1H NMR (400 MHz, DMSO-d6) δ 10.83 (s, 1H), 10.05 (s, 1H), 9.56 (s, 1H), 7.99 (dt, J = 7.0, 1.4 Hz, 2H), 7.66 - 7.54 (m, 5H), 7.39 (d, J = 7.7 Hz, 1H), 7.32 - 7.25 (m, 3H), 7.04 (t, J = 7.6 Hz, 1H), 6.96 (t, J = 7.4 Hz, 1H), 6.85 (br, 1H), 3.73 (d, J = 15.5 Hz, 1H), 3.47 (d, J = 9.4 Hz, 1H), 3.23 - 3.10 (m, 2H), 2.98 - 2.86 (m, 1H), 2.85 - 2.76 (m, 1H), 2.70 - 2.63 (m, 1H), 2.58 (td, J = 11.3, 3.9 Hz, 1H), 2.28 - 2.17 (m, 1H). 13 C NMR (100 MHz, DMSO-d6) δ 165.87, 165.44, 136.68, 135.42, 134.59, 133.05, 132.43, 131.84, 131.77, 131.51, 130.12, 129.12, 128.01, 126.92, 126.33, 126.08, 126.04, 125.82, 121.06, 118.86, 118.10, 111.45, 107.05, 55.02, 53.67, 52.11, 31.59, 21.69. HRMS (ESI) m / z: calcd for (C 29 H 27 O2N4+H) + ,463.2129; found: 463.2119.

[0074]

[0075] 10. Preparation of N-(3-benzoylamino phenyl)-l,4,6,7,12,12b-hexahydroindolo[2,3- a] quinoline-3-carboxamide (Compound I-7)

[0076] Prepared in the same manner as described in Example 2, steps 1-4, except that in step 2 one of the starting materials was N-Boc-1,3-phenylenediamine and in step 4 one of the starting materials was benzoic acid, to give 180 mg of a white solid in 61% yield. 1H NMR (400 MHz, DMSO-d6) δ 10.86 (s, 1H), 10.29 (s, 1H), 9.86 (s, 1H), 8.24 (t, J = 2.1 Hz, 1H), 8.01 - 7.94 (m, 2H), 7.61 - 7.51 (m, 3H), 7.47 - 7.39 (m, 3H), 7.32 (d, J = 8.1 Hz, 1H), 7.28 (t, J = 8.2 Hz, 1H), 7.05 (ddd, J = 8.1, 7.0, 1.3 Hz, 1H), 6.97 (td, J = 7.4, 1.1 Hz, 1H), 6.84 (br, 1H), 3.76 (d, J = 18.5 Hz, 1H), 3.49 (d, J = 12.4 Hz, 1H), 3.23 - 3.12 (m, 2H), 2.99 - 2.91 (m, 1H), 2.88 - 2.78 (m, 1H), 2.72 - 2.65 (m, 1H), 2.60 (td, J = 11.3, 3.9 Hz, 1H), 2.29 - 2.21 (m, 1H). 13 CNMR (100 MHz, DMSO-d6) δ 165.99, 165.62, 139.80, 139.76, 136.69, 135.55, 135.44, 133.53, 132.01, 130.95, 128.98, 128.83, 128.17, 126.96, 121.04, 118.86, 118.11, 116.26, 113.15, 111.45, 107.08, 55.19, 53.76, 52.16, 31.49, 21.74. HRMS (ESI) m / z: calcd for (C 29 H 27 O2N4+H) + ,463.2129; found: 463.2118.

[0077]

[0078] 11. N-(4-benzoylamino phenyl)-l,4,6,7,12,12b-hexahydroindolo[2,3- a]quinoline-3-carboxamide (Compound I-8) was prepared according to the same procedure as in Example 2, steps 1-4, except that in step 2 one of the starting materials was N-Boc-1,4-phenylenediamine and in step 4 one of the starting materials was benzoic acid, to give 130 mg of a white solid in 59% yield. 1HNMR (400 MHz, DMSO-d6) δ 10.90 (s, 1H), 10.24 (s, 1H), 9.84 (s, 1H), 7.96 (dt, J = 6.9, 1.5 Hz, 2H), 7.76 - 7.65 (m, 4H), 7.62 - 7.49 (m, 3H), 7.41 (d, J = 7.7 Hz, 1H), 7.33 (d, J = 8.0 Hz, 1H), 7.06 (t, J = 8.2 Hz, 1H), 6.98 (t, J = 8.0 Hz, 1H), 6.85 (br, 1H), 3.81 (d, J = 16.4 Hz, 1H), 3.59 (s, 1H), 3.30 - 3.19 (m, 2H), 2.98 (d, J = 17.6 Hz, 1H), 2.91 - 2.78 (m, 1H), 2.76 - 2.60 (m, 2H), 2.28 (t, J = 13.9 Hz, 1H). 13 CNMR (100 MHz, DMSO-d6) δ 165.76, 165.27, 136.72, 135.46, 135.38, 135.18, 131.95, 130.67, 128.85, 128.07, 126.88, 121.12, 120.83, 118.93, 118.16, 111.50, 107.01, 55.22, 53.56, 52.12, 31.25, 21.53. HRMS (ESI) m / z: calcd for (C 29 H 27 O2N4+H) + ,463.2129; found: 463.2116.

[0079]

[0080] 12. Preparation of N-(2-(2,3-dihydrobenzofuran-5-carboxamido)phenyl)- 1,4,6,7,12,12b-hexahydroindolo[2,3-a]quinoline-3-carboxamide (Compound I-9)

[0081] Prepared in the same manner as in Example 2, steps 1-4, except that one of the starting materials was 2,3-dihydrobenzofuran-5-carboxylic acid, to give a white solid 172 mg, yield: 61%. 1 H NMR (400 MHz, DMSO-d6) δ 10.86 (s, 1H), 9.96 (s, 1H), 9.45 (s, 1H), 7.87 (dd, J = 8.6, 1.6 Hz, 1H), 7.82 (d, J = 8.0 Hz, 1H), 7.76 - 7.65 (m, 4H), 7.62 - 7.49 (m, 3H), 7.41 (d, J = 7.7 Hz, 1H), 7.33 (d, J = 8.0 Hz, 1H), 7.06 (t, J = 8.2 Hz, 1H), 6.98 (t, J = 8.0 Hz, 1H), 6.85 (br, 1H), 3.81 (d, J = 16.4 Hz, 1H), 3.59 (s, 1H), 3.30 - 3.19 (m, 2H), 2.98 (d, J = 17.6 Hz, 1H), 2.91 - 2.78 (m, 1H), 2.76 - 2.60 (m, 2H), 2.28 (t, J = 13.9 Hz, 1H).

[0082] 2.0, 0.9 Hz, 1 H), 7.81 (dd, J = 8.4, 2.0 Hz, 1 H), 7.75 - 7.66 (m, 2 H), 7.58 (dd, J = 6.9, 2.4 Hz, 1 H), 7.35 - 7.27 (m, 2 H), 7.25 (td, J = 7.2, 2.6 Hz, 1 H), 7.16 - 7.08 (m, 2 H), 6.85 (d, J = 8.4 Hz, 1 H), 4.49 (td, J = 4.9,

[0083] 1.7 Hz, 1 H), 4.41 (t, J = 4.2 Hz, 2 H), 3.98 (dq, J = 13.5, 1.0 Hz, 1 H), 3.85 (dq, J = 13.7, 1.1 Hz, 1 H), 3.29 - 3.13 (m, 4 H), 2.99 - 2.89 (m, 2 H), 2.73-2.48 (m, 2 H). 13 C NMR (100 MHz, DMSO-d6) δ 169.40, 167.01, 163.55, 137.59, 136.45, 133.28, 132.39, 131.42, 128.77, 128.75, 127.90, 126.80, 126.74, 124.17, 123.89, 123.82, 122.15, 121.85, 121.27, 120.39, 119.08, 111.52, 109.96, 108.04, 71.56, 55.21, 53.56, 52.11, 31.63, 28.80, 21.67.

[0084] 13. Preparation of N-(2-benzamidinoethyl)-l,4,6,7,12,12-hexahydroindolo[2,3- a]quinoline-3-carboxamide (Compound I-10)

[0085] Prepared in the same manner as described in Example 2, steps 1-4, except that in step 2 one of the starting materials was N-Boc-1,2-ethanediamine and in step 4 one of the starting materials was benzoic acid, to give 200 mg of a white solid in 57% yield. 1H NMR (400 MHz, DMSO-d6) δ 10.83 (s, 1H), 8.57 (t, J = 5.4 Hz, 1H), 8.08 (t, J = 5.5 Hz, 1H), 7.90 - 7.82 (m, 2H), 7.54 - 7.44 (m, 3H), 7.39 (d, J = 7.8 Hz, 1H), 7.30 (d, J = 8.0 Hz, 1H), 7.03 (t, J = 7.5 Hz, 1H), 6.96 (t, J = 7.4 Hz, 1H), 6.64 (s, 1H), 3.66 (d, J = 16.4 Hz, 1H), 3.42 (d, J = 9.2 Hz, 1H), 3.35 - 3.30 (m, 4H), 3.15 (dd, J = 11.2, 5.0 Hz, 1H), 3.09 - 3.03 (m, 1H), 2.89 - 2.77 (m, 2H), 2.69 - 2.64 (m, 1H), 2.62 - 2.50 (m, 1H), 2.23 - 2.11 (m, 1H). 13 C NMR (100 MHz, DMSO-d6) δ 166.95, 166.69, 136.66, 135.55, 134.98, 133.13, 131.61, 129.61, 128.74, 127.67, 126.92, 121.02, 118.84, 118.08, 111.44, 107.01, 55.19, 53.81, 52.18, 39.63, 39.14, 31.38, 21.69. HRMS (ESI) m / z: calcd for (C 25 H 27 O2N4+H) + , 415.2129; found: 415.2116.

[0086]

[0087] 14. Preparation of N-(3-benzamidopropyl)-l,4,6,7,12,12-hexahydroindolo[2,3- a]quinoline-3-carboxamide (Compound I-11)

[0088] Prepared in the same manner as described in Example 2, steps 1-4, except that in step 2 one of the starting materials was N-Boc-1,3-propanediamine and in step 4 one of the starting materials was benzoic acid, to give 200 mg of a white solid in 66% yield. 1H NMR (400 MHz, DMSO-d6) δ 11.31 (s, 1H), 8.53 (t, J = 5.7 Hz, 1H), 8.39 (t, J = 5.8 Hz, 1H), 7.85 (dt, J = 6.9, 1.5 Hz, 2H), 7.55 - 7.44 (m, 4H), 7.40 (d, J = 8.1 Hz, 1H), 7.15 (t, J = 7.0 Hz, 1H), 7.05 (t, J = 7.5 Hz, 1H), 6.87 (br, 1H), 4.83 (s, 1H), 4.36 (d, J = 16.4 Hz, 1H), 4.09 - 3.87 (m, 2H), 3.62 - 3.41 (m, 2H), 3.36 - 3.20 (m, 4H), 3.08 - 3.00 (m, 2H), 2.63 - 2.52 (m, 1H), 1.80 - 1.67 (m, 2H). 13 C NMR (100 MHz, DMSO-d6) δ 166.71, 164.81, 137.04, 135.03, 131.59, 129.05, 128.76, 127.59, 125.99, 122.53, 119.70, 118.75, 112.01, 106.19, 55.77, 51.76, 51.22, 37.46, 37.20, 29.60, 28.24, 19.04. HRMS (ESI) m / z: calcd for (C 26 H 29 O2N4+H) + , 429.2285; found: 429.2274.

[0089]

[0090] 15. Preparation of (4-benzoylpiperazin-l-yl)(l,4,6,7,12,12b-hexahydroindolo[2,3- a]quinolin-3-yl)methanone (Compound I-12)

[0091] Prepared in the same manner as described in Example 2, steps 1-4, except that in step 2 one of the starting materials was N-Boc-piperazine and in step 4 one of the starting materials was benzoic acid, to give 149 mg of a white solid in 43% yield. 1H NMR (400 MHz, DMSO-d6) δ 10.83 (s, 1H), 7.48 - 7.41 (m, 5H), 7.39 (d, J = 7.8 Hz, 1H), 7.31 (d, J = 8.0 Hz, 1H), 7.04 (t, J = 7.6 Hz, 1H), 6.96 (t, J = 7.9 Hz, 1H), 5.98 (s, 1H), 3.72 - 3.43 (m, 10H), 3.20 - 3.08 (m, 2H), 2.87 - 2.74 (m, 2H), 2.70 - 2.61 (m, 1H), 2.59 - 2.52 (m, 1H), 2.21 - 2.09 (m, 1H). 13 C NMR (100 MHz, DMSO-d6) δ 169.70, 168.94, 136.67, 136.12, 135.54, 132.73, 130.15, 128.94, 127.50, 126.95, 126.08, 121.03, 118.86, 118.09, 111.47, 106.98, 55.19, 54.65, 51.94, 47.64, 42.23, 30.80, 21.63. HRMS (ESI) m / z: calcd for (C 27 H 29 O2N4+H) + , 441.2285; found: 441.2273.

[0092]

[0093] 16. Preparation of N-(2-benzamidinophenyl)-9-methyl-1,4,6,7,12,12b- hexahydroindolo[2,3-a]quinoline-3-carboxamide (Compound 1-13)

[0094] Prepared in the same manner as described in Example 2, steps 1-4, except that in step 1 the starting material was replaced by compound IV-2 and in step 4 the starting material was replaced by benzoic acid, to give 192 mg of a white solid, yield: 72%. 1H NMR (400 MHz, DMSO-d6) δ 10.79 (s, 1H), 10.16 (s, 1H), 9.47 (s, 1H), 7.98 - 7.92 (m, 2H), 7.75 - 7.63 (m, 2H), 7.55 - 7.43 (m, 3H), 7.39 - 7.22 (m, 3H), 7.10-7.03 (m, 2H), 4.44 (td, J=5.0, 1.9 Hz, 1H), 3.95 (dq, J=13.6, 1.1 Hz, 1H), 3.87 (dq, J=13.8, 1.3 Hz, 1H), 3.26-3.19 (m, 2H), 3.00 - 2.95 (m, 2H), 2.76 (dt, J=14.1, 4.9 Hz, 1H), 2.54 (dt, J=14.1, 4.9 Hz, 1H). 13 C NMR (101 MHz, DMSO-d6) δ 169.34, 166.87, 137.58, 136.09, 134.59, 133.14, 132.88, 132.37, 131.41, 130.19, 129.13, 127.64, 126.31, 125.87, 124.18, 123.82, 122.13, 121.85, 121.27, 119.59, 109.78, 107.51, 55.13, 53.65, 52.11, 31.66, 21.48, 20.29.

[0095]

[0096] 17. Preparation of N-(2-(3-methoxy-4-methylbenzamido)phenyl)-9-methyl- 1,4,6,7,12,12b-hexahydroindolo[2,3-a]quinoline-3-carboxamide (Compound 1-14)

[0097] Prepared in the same manner as in Example 2, steps 1-4, except that in step 1 the starting material was replaced by compound IV-2 and in step 4 the starting material was replaced by 3-methoxy-4-methylbenzoic acid, to give 210 mg of a white solid in 72% yield. 1H NMR (400 MHz, DMSO-d6) δ 10.77 (s, 1H), 9.67 (s, 1H), 7.75 - 7.67 (m, 1H), 7.71 - 7.63 (m, 2H), 7.39 - 7.18 (m, 5H), 7.10-7.03 (m, 2H), 4.49 (td, J=4.9, 1.7 Hz, 1H), 3.97 (dq, J=13.5, 1.0 Hz, 1H), 3.87 - 3.81 (m, 1H), 3.82 (s, 2H), 3.30-3.17 (m, 2H), 3.00 - 2.87 (m, 2H), 2.73 (dt, J=14.1, 4.9 Hz, 1H), 2.48 (dt, J=14.1, 4.9 Hz, 1H), 2.40 (d, J=0.9 Hz, 3H), 2.16 (s, 1H). 13 C NMR (100 MHz, DMSO-d6) δ 165.90, 165.45, 136.68, 135.42, 134.60, 133.35, 132.45, 131.85, 131.77, 131.51, 130.14, 129.15, 128.00, 126.90, 126.35, 126.05, 126.01, 125.82, 121.04, 118.86, 118.11, 111.45, 107.07, 55.01, 53.67, 52.11, 31.60, 21.68, 16.46.

[0098]

[0099] 18. Preparation of N-(2-(3-fluorobenzamido)phenyl)-l,4,6,7,12,12b-hexahydroindolo[2,3- a]quinoline-3-carboxamide (Compound 1-15)

[0100] In a 100 mL flask was added 3-fluorobenzoic acid (55 mg, 0.39 mmol), N2-protected, to the flask was added DCM 5 mL and a drop of DMF, oxalyl chloride (67 μL, 0.78 mmol) was added dropwise slowly under ice bath, the reaction was allowed to proceed for 1 h at room temperature, after TLC indicated the reaction was complete, the reaction solvent was evaporated under reduced pressure, then compound N-(2-aminophenyl)-l,4,6,7,12,12b-hexahydroindolo[2,3- a]quinoline-3-carboxamide (154 mg, 0.43 mmol) prepared in the same manner as in Example 2, steps 1-3, and 10 mL DCM were added, TEA (150 μL, 1.08 mmol) was added dropwise slowly under ice bath, the reaction was allowed to proceed for 12 h at room temperature, after the reaction was complete, the reaction solvent was evaporated under reduced pressure, the reaction mixture was extracted with saturated NaHCO3solution and EA three times, washed with saturated NaCl once, the organic phase was collected, dried over anhydrous MgSO4, and finally purified by column chromatography (DCM:MeOH = 50: 1) to give 125 mg of white solid in 66% yield. 1 H NMR (400 MHz, DMSO-d6) δ 10.84 (s, 1H), 10.12 (s, 1H), 9.53 (s, 1H), 7.84 (d, J = 7.7 Hz, 1H), 7.79 (dt, J = 9.6, 2.1 Hz, 1H), 7.67 - 7.57 (m, 3H), 7.48 (td, J = 8.3, 2.2 Hz, 1H), 7.40 (d, J = 7.8 Hz, 1H), 7.31 (d, J = 7.9 Hz, 1H), 7.30 - 7.25 (m, 2H), 7.05 (ddd, J = 8.2, 7.0, 1.3 Hz, 1H), 6.97 (t, J = 6.9 Hz, 1H), 6.86 (br, 1H), 3.75 (d, J = 16.3 Hz, 1H), 3.48 (d, J = 12.2 Hz, 1H), 3.23 - 3.11 (m, 2H), 2.98 - 2.89 (m, 1H), 2.86 - 2.77 (m, 1H), 2.71 - 2.65 (m, 1H), 2.59 (td, J = 11.3, 3.9 Hz, 1H), 2.28 - 2.18 (m, 1H). 13C NMR (100 MHz, DMSO-d6) δ 165.46, 164.60, 163.70, 161.27, 137.12, 137.05, 136.69, 135.38, 133.05, 131.96, 131.79, 131.33, 131.26, 126.93, 126.45, 126.26, 126.10, 125.79, 124.22, 124.19, 121.07, 119.37, 119.16, 118.88, 118.11, 115.11, 114.88, 111.47, 107.05, 55.05, 53.68, 52.13, 31.57, 21.68. HRMS (ESI) m / z: calcd for (C 29 H 26 O2N4F+H) + ,481.2034; found: 481.2021.

[0101]

[0102] 19. Preparation of N-(2-(3,4-difluorobenzamido)phenyl)-l,4,6,7,12,12b- hexahydroindolo[2,3-a]quinoline-3-carboxamide (Compound 1-17)

[0103] Prepared in the same manner as described in Example 2 for 18, except that the starting material was replaced by 3,4-difluorobenzoic acid, to give 103 mg of white solid, yield 53%. 1 H NMR (400 MHz, DMSO-d6) δ 10.84 (s, 1H), 10.13 (s, 1H), 9.49 (s, 1H), 8.04 (ddd, J = 11.4, 7.7, 2.2 Hz, 1H), 7.92 - 7.83 (m, 1H), 7.71 - 7.54 (m, 3H), 7.39 (d, J = 7.8 Hz, 1H), 7.31 (d, J = 7.9 Hz, 1H), 7.29 - 7.25 (m, 2H), 7.04 (ddd, J = 8.1, 7.0, 1.3 Hz, 1H), 6.96 (ddd, J = 8.1, 7.0, 1.1 Hz, 1H), 6.84 (br, 1H), 3.78 - 3.68 (m, 1H), 3.51 - 3.43 (m, 1H), 3.23 - 3.11 (m, 2H), 2.97 - 2.88 (m, 1H), 2.86 - 2.76 (m, 1H), 2.72 - 2.64 (m, 1H), 2.58 (td, J = 11.3, 4.0 Hz, 1H), 2.28 - 2.18 (m, 1H). 13C NMR (100 MHz, DMSO-d6) δ 165.47, 163.78, 153.48, 153.35, 150.98, 150.85, 148.52, 148.39, 136.69, 135.42, 133.09, 132.23, 132.19, 132.14, 132.05, 131.75, 131.15, 126.93, 126.50, 126.30, 126.10, 125.74, 125.65, 125.62, 125.58, 125.55, 121.05, 118.86, 118.43, 118.25, 118.10, 117.75, 117.57, 111.45, 107.05, 55.05, 53.70, 52.12, 31.58, 21.70. HRMS (ESI) m / z: calcd for (C 29 H 25 O2N4F2+H) + , 499.1940; found: 499.1928.

[0104]

[0105] 20. Preparation of N-(2-(3-fluoro-4-methoxybenzamido)phenyl)-1,4,6,7,12,12b- hexahydroindolo[2,3-a]quinoline-3-carboxamide (Compound 1-18)

[0106] Prepared in the same manner as described in Example 2 for 18, except that the starting material was replaced by 3-fluoro-4-methoxybenzoic acid, to give 124 mg of white solid in 62% yield. 1 H NMR (400 MHz, DMSO-d6) δ 10.81 (s, 1H), 9.98 (s, 1H), 9.49 (s, 1H), 7.85 - 7.77 (m, 2H), 7.61 - 7.53 (m, 2H), 7.37 - 7.20 (m, 5H), 7.01 (ddd, J = 8.1, 7.0, 1.3 Hz, 1H), 6.93 (td, J = 7.4, 1.1 Hz, 1H), 6.80 (br, 1H), 3.89 (s, 3H), 3.70 (d, J = 16.3 Hz, 1H), 3.44 (d, J = 13.0 Hz, 1H), 3.20 - 3.08 (m, 2H), 2.93 - 2.84 (m, 1H), 2.82 - 2.73 (m, 1H), 2.67 - 2.61 (m, 1H), 2.55 (td, J = 11.2, 3.9 Hz, 1H), 2.25 - 2.14 (m, 1H). 13C NMR (100 MHz, DMSO-d6) δ 165.38, 164.33, 152.61, 150.74, 150.63, 150.18, 136.69, 135.40, 133.06, 131.87, 131.79, 131.37, 127.01, 126.96, 126.93, 126.38, 126.08, 126.02, 125.75, 125.28, 125.24, 121.07, 118.87, 118.11, 115.79, 115.59, 113.95, 111.46, 107.04, 56.76, 55.04, 53.67, 52.13, 31.59, 21.69. HRMS (ESI) m / z: calcd for (C 30 H 28 O3N4F+H) + , 511.2140; found: 511.2127.

[0107]

[0108] 21. Preparation of N-(2-(3-fluoro-4-methoxybenzamido)phenyl)-1,4,6,7,12,12b- hexahydroindolo[2,3-a]quinoline-3-carboxamide (Compound 1-19)

[0109] Prepared in the same manner as described in Example 2 for 18, except that the starting material was replaced by 3-trifluoromethylbenzoic acid, to give 145 mg of white solid in 70% yield. 1 H NMR (400 MHz, DMSO-d6) δ 10.80 (s, 1H), 10.23 (s, 1H), 9.50 (s, 1H), 8.28 (s, 1H), 8.24 (d, J = 7.9 Hz, 1H), 7.95 (d, J = 7.8 Hz, 1H), 7.77 (t, J = 7.8 Hz, 1H), 7.66 - 7.54 (m, 2H), 7.36 (d, J = 7.7 Hz, 1H), 7.29 - 7.22 (m, 3H), 7.01 (ddd, J = 8.1, 7.0, 1.3 Hz, 1H), 6.93 (td, J = 7.4, 1.1 Hz, 1H), 6.83 (br, 1H), 3.70 (d, J = 14.7 Hz, 1H), 3.43 (d, J = 13.2 Hz, 1H), 3.19 - 3.05 (m, 2H), 2.92 - 2.84 (m, 1H), 2.82 - 2.72 (m, 1H), 2.67 - 2.60 (m, 1H), 2.53 (td, J = 11.3, 4.0 Hz, 1H), 2.24 - 2.12 (m, 1H). 13C NMR (100 MHz, DMSO-d6) δ 165.47, 164.46, 136.69, 135.75, 135.39, 132.98, 132.25, 132.01, 131.87, 131.28, 130.40, 130.25, 129.92, 129.60, 129.28, 128.86, 128.82, 128.50, 126.92, 126.45, 126.31, 126.16, 125.82, 124.73, 124.69, 124.66, 124.62, 123.09, 121.07, 118.87, 118.10, 111.46, 107.05, 55.04, 53.69, 52.13, 31.55, 21.68. HRMS (ESI) m / z: calcd for (C 30 H 26 O2N4F3+H) + , 531.2002; found: 531.1990.

[0110]

[0111] 22. Preparation of N-(2-(3-methoxy-4-nitrobenzamido)phenyl)-1,4,6,7,12,12b- hexahydroindolo[2,3-a]quinoline-3-carboxamide (Compound I-20)

[0112] Prepared in the same manner as described in Example 2 for 18, except that the starting material was replaced by 3-methoxy-4-nitrobenzoic acid, to give 138 mg of white solid in 66% yield. 1 H NMR (400 MHz, DMSO-d6) δ 10.80 (s, 1H), 10.18 (s, 1H), 9.52 (s, 1H), 8.02 (d, J = 8.3 Hz, 1H), 7.82 (s, 1H), 7.66 - 7.53 (m, 3H), 7.36 (d, J = 7.7 Hz, 1H), 7.29 - 7.23 (m, 3H), 7.00 (t, J = 7.4 Hz, 1H), 6.93 (t, J = 7.4 Hz, 1H), 6.83 (br, 1H), 3.99 (s, 3H), 3.71 (d, J = 16.0 Hz, 1H), 3.43 (dd, J = 10.7, 3.5 Hz, 1H), 3.19 - 3.07 (m, 2H), 2.92 - 2.83 (m, 1H), 2.81 - 2.72 (m, 1H), 2.63 (dd, J = 15.0, 3.8 Hz, 1H), 2.54 (td, J = 11.3, 4.0 Hz, 1H), 2.24 - 2.13 (m, 1H). 13C NMR (100 MHz, DMSO-d6) δ 165.57, 164.23, 152.08, 141.46, 139.96, 136.67, 135.44, 133.05, 131.96, 131.87, 131.14, 126.93, 126.36, 126.19, 125.82, 125.59, 121.06, 120.16, 118.86, 118.10, 113.88, 111.45, 107.06, 57.35, 55.06, 53.70, 52.12, 31.59, 21.71. HRMS (ESI) m / z: calcd for (C 30 H 28 O5N5F+H) + , 538.2085; found: 538.2072.

[0113]

[0114] 23. Preparation of N-(2-(3-nitrobenzamido)phenyl)-l,4,6,7,12,12b-hexahydroindolo[2,3- a]quinoline-3-carboxamide (Compound 1-21)

[0115] Prepared in the same manner as described in Example 2 for 18, except that the starting material was replaced with m-nitrobenzoic acid, to give 109 mg of white solid in 55% yield. 1 H NMR (400 MHz, DMSO-d6) δ 10.84 (s, 1H), 10.36 (s, 1H), 9.56 (s, 1H), 8.81 (t, J = 2.0 Hz, 1H), 8.46 (ddd, J = 8.2, 2.4, 1.0 Hz, 1H), 8.41 (dt, J = 7.8, 1.3 Hz, 1H), 7.86 (t, J = 8.0 Hz, 1H), 7.69 - 7.59 (m, 2H), 7.39 (d, J = 7.8 Hz, 1H), 7.33 - 7.26 (m, 3H), 7.04 (ddd, J = 8.1, 7.1, 1.2 Hz, 1H), 6.96 (ddd, J = 8.0, 7.4, 1.1 Hz, 1H), 6.85 (br, 1H), 3.76 (dd, J = 16.5, 2.5 Hz, 1H), 3.48 (dd, J = 9.8, 3.5 Hz, 1H), 3.23 - 3.12 (m, 2H), 2.97 - 2.89 (m, 1H), 2.85 - 2.77 (m, 1H), 2.70 - 2.64 (m, 1H), 2.58 (td, J = 11.2, 3.9 Hz, 1H), 2.30 - 2.16 (m, 1H). 13C NMR (100 MHz, DMSO-d6) δ 165.60, 163.94, 148.31, 136.75, 136.40, 135.51, 134.65, 133.10, 132.15, 131.93, 131.23, 130.90, 126.99, 126.89, 126.61, 126.44, 126.27, 125.86, 122.93, 121.12, 118.93, 118.17, 111.53, 107.12, 55.12, 53.79, 52.20, 31.65, 21.77. HRMS (ESI) m / z: calcd for (C 29 H 26 O4N5+H) + ,508.1979; found: 508.1967.

[0116]

[0117] 24. Preparation of N-(2-(3-methoxy-4-trifluoromethylbenzamido)phenyl)- 1,4,6,7,12,12b-hexahydroindolo[2,3-a]quinoline-3-carboxamide (Compound I-22)

[0118] Prepared in the same manner as described in Example 2 for 18, except that the starting material was replaced by 3-methoxy-4-trifluoromethylbenzoic acid, to give 109 mg of white solid in 50% yield. 1 H NMR (400 MHz, DMSO-d6) δ 10.81 (s, 1H), 10.15 (s, 1H), 9.54 (s, 1H), 7.78 (d, J = 8.1 Hz, 1H), 7.73 (s, 1H), 7.65 - 7.60 (m, 2H), 7.60 - 7.54 (m, 1H), 7.36 (d, J = 7.7 Hz, 1H), 7.29 - 7.23 (m, 3H), 7.00 (t, J = 8.1 Hz, 1H), 6.93 (t, J = 6.9 Hz, 1H), 6.83 (br, 1H), 3.97 (s, 3H), 3.71 (d, J = 16.3 Hz, 1H), 3.50 - 3.35 (m, 1H), 3.18 - 3.09 (m, 2H), 2.92 - 2.85 (m, 1H), 2.81 - 2.73 (m, 1H), 2.67 - 2.60 (m, 1H), 2.59 - 2.50 (m, 1H), 2.24 - 2.12 (m, 1H). 13CNMR (100 MHz, DMSO-d6) δ 165.62, 164.74, 157.59, 140.44, 136.74, 136.59, 135.46, 135.32, 133.08, 131.96, 131.30, 127.82, 127.77, 127.72, 126.99, 126.95, 126.37, 126.25, 126.16, 125.91, 121.14, 119.95, 118.94, 118.18, 112.33, 111.52, 107.12, 56.90, 55.12, 53.73, 52.18, 31.63, 21.74. HRMS (ESI) m / z: calcd for (C 31 H 28 O3N4F3+H) + ,561.2108; found: 561.2094.

[0119]

[0120] 25. Preparation of N-(2-(3-methyl-4-nitrobenzamido)phenyl)-1,4,6,7,12,12b- hexahydroindolo[2,3-a]quinoline-3-carboxamide (Compound 1-23)

[0121] Prepared in the same manner as described in Example 2 for 18, except that the starting material was replaced by 3-methyl-4-nitrobenzoic acid, to give 108 mg of white solid in 54% yield. 1 H NMR (400 MHz, DMSO-d6) δ 10.81 (s, 1H), 10.19 (s, 1H), 9.50 (s, 1H), 8.10 (d, J = 8.5 Hz, 1H), 8.00 (s, 1H), 7.93 (dd, J = 8.4, 2.0 Hz, 1H), 7.63 - 7.53 (m, 2H), 7.35 (dd, J = 7.8, 1.2 Hz, 1H), 7.29 - 7.22 (m, 3H), 7.00 (ddd, J = 8.2, 7.0, 1.3 Hz, 1H), 6.92 (ddd, J = 8.0, 7.0, 1.1 Hz, 1H), 6.82 (br, 1H), 3.70 (d, J = 16.3 Hz, 1H), 3.45 (d, J = 7.0 Hz, 1H), 3.17 - 3.08 (m, 2H), 2.93 - 2.84 (m, 1H), 2.80 - 2.73 (m, 1H), 2.67 - 2.58 (m, 2H), 2.55 (s, 3H), 2.24 - 2.14 (m, 1H). 13C NMR (100 MHz, DMSO-d6) δ 165.54, 164.48, 151.15, 138.79, 136.72, 135.42, 133.44, 133.03, 132.56, 132.01, 131.95, 131.26, 127.03, 126.96, 126.48, 126.29, 125.93, 125.25, 121.15, 118.95, 118.19, 111.53, 107.11, 55.10, 53.73, 52.18, 31.60, 21.71, 20.00. HRMS (ESI) m / z: calcd for (C 30 H 28 O4N5+H) + ,522.2136; found: 522.2120.

[0122]

[0123] 26. Preparation of N-(2-(4-nitrobenzamido)phenyl)-l,4,6,7,12,12b- hexahydroindolo[2,3-a]quinoline-3-carboxamide (Compound 1-24)

[0124] Prepared in the same manner as Example 2,18except that the starting material was replaced with 4-nitrobenzoic acid to give 130 mg of white solid in 67% yield. 1 H NMR (400 MHz, DMSO-d6) δ 10.79 (s, 1H), 10.27 (s, 1H), 9.50 (s, 1H), 8.39 - 8.34 (m, 2H), 8.19 - 8.14 (m, 2H), 7.63 - 7.57 (m, 2H), 7.35 (d, J = 7.8 Hz, 1H), 7.30 - 7.22 (m, 3H), 7.00 (ddd, J = 8.1, 7.0, 1.3 Hz, 1H), 6.92 (td, J = 7.4, 1.1 Hz, 1H), 6.82 (br, 1H), 3.70 (d, J = 16.3 Hz, 1H), 3.44 (dd, J = 10.8, 3.4 Hz, 1H), 3.20 - 3.07 (m, 2H), 2.92 - 2.84 (m, 1H), 2.83 - 2.72 (m, 1H), 2.67 - 2.60 (m, 1H), 2.55 (td, J = 11.2, 3.9 Hz, 1H), 2.24 - 2.15 (m, 1H). 13CNMR (100 MHz, DMSO-d6) δ 165.64, 164.38, 149.84, 140.55, 136.75, 135.50, 133.14, 132.15, 131.90, 131.16, 129.66, 127.00, 126.63, 126.50, 126.25, 125.85, 124.27, 121.12, 118.93, 118.17, 111.52, 107.12, 55.11, 53.75, 52.18, 31.65, 21.76. HRMS (ESI) m / z: calcd for (C 29 H 26 O4N5+H) + ,508.1979; found: 508.1967.

[0125]

[0126] 27. Preparation of N-(2-(4-trifluoromethylcarboxamido)phenyl)-l,4,6,7,12,12b- hexahydroindolo[2,3-a]quinoline-3-carboxamide (Compound I-25)

[0127] Prepared in the same manner as described in Example 2,18except that the starting material was replaced by 4-trifluoromethylbenzoic acid to give 120 mg of white solid in 60% yield. 1 H NMR (400 MHz, DMSO-d6) δ 10.80 (s, 1H), 10.20 (s, 1H), 9.50 (s, 1H), 8.14 (d, J = 8.1 Hz, 2H), 7.92 (d, J = 8.3 Hz, 2H), 7.65 - 7.53 (m, 2H), 7.36 (d, J = 7.7 Hz, 1H), 7.30 - 7.17 (m, 3H), 7.01 (ddd, J = 8.2, 7.1, 1.3 Hz, 1H), 6.93 (ddd, J = 8.0, 7.0, 1.1 Hz, 1H), 6.82 (br, 1H), 3.70 (d, J = 16.3 Hz, 1H), 3.49 - 3.41 (m, 1H), 3.19 - 3.09 (m, 2H), 2.93 - 2.85 (m, 1H), 2.82 - 2.73 (m, 1H), 2.67 - 2.60 (m, 1H), 2.55 (td, J = 10.0, 2.8 Hz, 1H), 2.25 - 2.15 (m, 1H). 13CNMR (100 MHz, DMSO-d6) δ 165.59, 164.84, 162.85, 138.64, 136.76, 135.45, 133.09, 132.32, 132.07, 132.00, 131.88, 131.27, 129.07, 126.99, 126.58, 126.40, 126.21, 126.17, 126.13, 126.09, 125.86, 125.80, 123.09, 121.14, 118.94, 118.17, 111.53, 107.11, 55.10, 53.72, 52.16, 31.61, 21.72. HRMS (ESI) m / z: calcd for (C 30 H 26 O2N4F3+H) + , 531.2002; found: 531.1987.

[0128]

[0129] 28. Preparation of N-(2-([l,l'-biphenyl]-3-carboxamide)phenyl)-l,4,6,7,12,12b- hexahydroindolo[2,3-a]quinoline-3-carboxamide (Compound I-26)

[0130] Prepared in the same manner as described in Example 2 for 18, except substituting [l,l'-biphenyl]-3-carboxylic acid as the starting material, to give 104 mg of a white solid in 51% yield. 1H NMR (400 MHz, DMSO-d6) δ 10.80 (s, 1H), 10.13 (s, 1H), 9.61 (s, 1H), 8.23 (t, J = 1.9 Hz, 1H), 7.95 (dt, J = 7.8, 1.3 Hz, 1H), 7.88 (ddt, J = 7.7, 1.9, 0.9 Hz, 1H), 7.76 - 7.72 (m, 2H), 7.71 - 7.66 (m, 1H), 7.62 (t, J = 7.8 Hz, 1H), 7.59 - 7.55 (m, 1H), 7.49 (t, J = 7.6 Hz, 2H), 7.37 (t, J = 7.4 Hz, 2H), 7.30 - 7.23 (m, 3H), 7.01 (ddd, J = 8.1, 7.1, 1.2 Hz, 1H), 6.93 (t, J = 7.4 Hz, 1H), 6.86 (br, 1H), 3.71 (d, J = 16.3 Hz, 1H), 3.45 - 3.39 (m, 1H), 3.19 - 3.10 (m, 1H), 3.07 - 3.01 (m, 1H), 2.93 - 2.85 (m, 1H), 2.80 - 2.70 (m, 1H), 2.65 - 2.57 (m, 1H), 2.55 - 2.48 (m, 1H), 2.25 - 2.13 (m, 1H). 13 C NMR (100 MHz, DMSO-d6) δ 165.69, 165.62, 141.01, 139.94, 136.76, 135.46, 135.39, 133.01, 132.13, 131.77, 131.69, 130.66, 129.95, 129.64, 128.47, 127.47, 127.42, 126.99, 126.28, 126.16, 126.13, 126.08, 125.98, 121.14, 118.95, 118.18, 111.54, 107.11, 55.09, 53.75, 52.19, 31.66, 21.74. HRMS (ESI) m / z: calcd for (C 35 H 31 O2N4+H) + , 539.2442; found: 539.2433.

[0131]

[0132] 29. Preparation of N-(2-benzylcarbamimidoyl-ethyl)-1,4,6,7,12,12b- hexahydroindolo[2,3-a]quinoline-3-carboxamide (Compound 1-27)

[0133] Prepared in the same manner as in example 2, steps 1-4, except that in step 2 one of the starting materials was tert-butyl (2-aminopropyl)carbamate and in step 4 one of the starting materials was 3- methoxybenzoic acid, to give a white solid, 169 mg, yield for the last step: 68%. 1 H NMR (400 MHz, DMSO-d6) δ 10.83 (s, 1H), 8.57 (t, J = 5.4 Hz, 1H), 8.08 (t, J = 5.5 Hz, 1H), 7.90 - 7.82 (m, 2H), 7.54 - 7.44 (m, 3H), 7.39 (d, J = 7.8 Hz, 1H), 7.30 (d, J = 8.0 Hz, 1H), 7.03 (t, J = 7.5 Hz, 1H), 6.96 (t, J = 7.4 Hz, 1H), 6.64 (s, 1H), 3.66 (d, J = 16.4 Hz, 1H), 3.42 (d, J = 9.2 Hz, 1H), 3.35 - 3.30 (m, 4H), 3.15 (dd, J = 11.2, 5.0 Hz, 1H), 3.09 - 3.03 (m, 1H), 2.89 - 2.77 (m, 2H), 2.69 - 2.64 (m, 1H), 2.62 - 2.50 (m, 1H), 2.23 - 2.11 (m, 1H). 13 C NMR (100 MHz, DMSO-d6) δ 166.95, 166.69, 136.66, 135.55, 134.98, 133.13, 131.61, 129.61, 128.74, 127.67, 126.92, 121.02, 118.84, 118.08, 111.44, 107.01, 55.19, 53.81, 52.18, 39.63, 39.14, 31.38, 21.69. HRMS (ESI) m / z: calcd for (C 25 H 27 O2N4+H) + ,415.2129; found: 415.2116.

[0134]

[0135] 30. Preparation of N-(2-(3-methoxybenzamido)ethyl)-l,4,6,7,12,12b-hexahydroindolo[2,3- a]quinoline-3-carboxamide (Compound I-28)

[0136] Prepared in the same manner as in example 2, steps 1-4, except that in step 2 one of the starting materials was tert-butyl (2-aminopropyl)carbamate and in step 4 one of the starting materials was 3- methoxybenzoic acid, to give a white solid, 169 mg, yield for the last step: 68%.1 H NMR (400 MHz, Chloroform-d) δ 8.45 (s, 1H), 7.67 (dd, J = 8.1, 2.1 Hz, 1H), 7.54 (dd, J = 6.9, 2.4 Hz, 1H), 7.40 - 7.32 (m, 3H), 7.32 - 7.27 (m, 1H), 7.23 (td, J = 7.2, 2.6 Hz, 1H), 7.13 (td, J = 7.0, 2.0 Hz, 1H), 7.03 - 6.90 (m, 3H), 4.49 (td, J = 4.9, 1.7 Hz, 1H), 3.91 (dq, J = 13.6, 1.0 Hz, 1H), 3.82 - 3.75 (m, 3H), 3.44 (dt, J = 5.3, 4.4 Hz, 2H), 3.35 (dt, J = 5.4, 4.4 Hz, 2H), 3.30-3.17 (m, 2H), 2.99 - 2.87 (m, 2H), 2.73 (dt, J = 13.9, 4.8 Hz, 1H), 2.48 (dt, J = 13.9, 4.8 Hz, 1H). 13 C NMR (100 MHz, Chloroform-d) δ 171.55, 167.00, 160.27, 136.84, 136.40, 135.11, 133.27, 129.27, 126.79, 123.86, 122.37, 121.18, 120.39, 119.08, 117.86, 112.33, 111.52, 108.04, 55.03, 53.66, 52.11, 49.93, 39.56, 39.55, 31.61, 21.43.

[0137]

[0138] 31. Preparation of N-(2-(3-methoxy-4-methylbenzoylamino)ethyl)-l,4,6,7,12,12b- hexahydroindolo[2,3-a]quinoline-3-carboxamide (Compound 1-29)

[0139] Prepared in the same manner as in Example 2, steps 1-4, except that in step 2 one of the starting materials was tert-butyl (2-aminoethyl)carbamate and in step 4 one of the starting materials was 3-methoxy-4-methylbenzoic acid, to give a white solid 162 mg, yield for the last step: 63%. 1H NMR (400 MHz, Chloroform-d) δ 8.55 (s, 1H), 7.79 (dd, J = 8.2, 2.0 Hz, 1H), 7.56 (dd, J = 6.9, 2.4 Hz, 1H), 7.45 (t, J = 5.2 Hz, 1H), 7.32 - 7.28 (m, 1H), 7.27 - 7.21 (m, 2H), 7.20 - 7.10 (m, 2H), 7.07 (dt, J = 4.9, 1.8 Hz, 1H), 6.98 (t, J = 5.6 Hz, 1H), 4.50 (td, J = 4.9, 1.7 Hz, 1H), 3.99 (dq, J = 13.5, 1.0 Hz, 1H), 3.84 - 3.70 (m, 3H), 3.44 (dt, J = 5.5, 4.5 Hz, 2H), 3.36 (dt, J = 5.4, 4.4 Hz, 2H), 3.33-3.17 (m, 2H), 2.95 - 2.84 (m, 2H), 2.73 (dt, J = 13.9, 5.0 Hz, 1H), 2.50 (dt, J = 13.9, 4.8 Hz, 1H), 2.20 (s, 1H). 13 C NMR (100 MHz, Chloroform-d) δ 171.45, 167.04, 157.92, 136.94, 136.45, 133.29, 132.28, 130.94, 130.06, 126.88, 123.80, 122.47, 121.85, 120.55, 119.48, 111.62, 109.57, 108.24, 55.17, 53.66, 52.42, 51.93, 39.56, 39.55, 31.68, 21.76, 16.69.

[0140]

[0141] 32. Preparation of N-(2-(3,4-difluorobenzamido)ethyl)-l,4,6,7,12,12b- hexahydroindolo[2,3-a]quinoline-3-carboxamide (Compound I-30)

[0142] 3243. Prepared in the same manner as in Example 2, 18, except that the starting materials were replaced with 3,4-difluorobenzoic acid and Compound N-(2- aminoethyl)-l,4,6,7,12,12b-hexahydroindolo[2,3-a]quinoline-3-carboxamide prepared in the same manner as in Example 2, Steps 1-3, to give 74 mg of white solid in 42% yield. 1H NMR (400 MHz, Chloroform-d) δ 8.65 (s, 1H), 7.92 - 7.77 (m, 2H), 7.64 (dd, J = 6.9, 2.4 Hz, 1H), 7.50 (t, J = 5.5 Hz, 1H), 7.47 (ddd, J = 10.3, 9.3, 4.8 Hz, 1H), 7.32 - 7.29 (m, 1H), 7.23 (td, J = 7.5, 2.8 Hz, 1H), 7.16 (td, J = 7.5, 2.8 Hz, 1H), 7.13 (td, J = 5.0, 1.8 Hz, 1H), 7.06 (t, J = 5.5 Hz, 1H), 4.50 (td, J = 5.2, 1.7 Hz, 1H), 3.95 (dq, J = 13.5, 1.5 Hz, 1H), 3.80 (dq, J = 13.6, 1.3 Hz, 1H), 3.47 (dt, J = 5.3, 4.8 Hz, 2H), 3.38 (dt, J = 5.3, 4.7 Hz, 2H), 3.33 - 3.17 (m, 2H), 3.09 - 2.99 (m, 2H), 2.83 (dtt, J = 13.9, 4.8, 0.9 Hz, 1H), 2.58 (dt, J = 13.9, 4.8 Hz, 1H). 13 C NMR (100 MHz, Chloroform-d) δ 171.50, 166.50, 166.41, 156.02, 155.90, 154.08, 153.90, 151.60, 151.43, 149.67, 149.52, 136.85, 136.40, 133.25, 130.40, 130.35, 130.38, 130.35, 126.79, 125.25, 125.22, 125.20, 125.18, 123.84, 122.38, 120.39, 119.08, 118.21, 118.18, 118.02, 117.99, 117.85, 117.81, 117.68, 117.64, 111.50, 108.04, 55.15, 53.42, 52.03, 39.56, 39.55, 31.71, 21.73.

[0143]

[0144] 33. Preparation of N-(2-(3-trifluoromethylbenzamido)ethyl)-l,4,6,7,12,12b- hexahydroindolo[2,3-a]quinoline-3-carboxamide (Compound I-31)

[0145] Prepared in the same manner as in Example 2, 18 except substituting 3- trifluoromethylbenzoic acid and compound N-(2-aminoethyl)-l,4,6,7,12,12b- hexahydroindolo[2,3-a]quinoline-3-carboxamide prepared in the same manner as in Example 2, steps 1-3, in the last step, in white solid 77 mg, yield: 41%. 1 H NMR (400 MHz, Chloroform-d) δ 8.49 (s, 1H), 8.14 (t, J = 2.1 Hz, 1H), 7.71 (ddd, J = 7.5, 2.2, 1.3 Hz, 1H), 7.68 (ddd, J = 10.5, 2.2, 1.2 Hz, 1H), 7.60 (dd, J = 10.7, 7.6 Hz, 1H), 7.50 (dd, J = 6.9, 2.5 Hz, 1H), 7.46 - 7.20 (m, 3H), 7.03 (td, J = 7.3, 2.1 Hz, 1H), 7.01 (td, J = 4.9, 1.8 Hz, 1H), 6.93 (t, J = 5.6 Hz, 1H), 4.41 (td, J = 4.9, 1.5 Hz, 1H), 3.71 (dq, J = 13.5, 1.0 Hz, 1H), 3.69 (dq, J = 13.6, 1.1 Hz, 1H), 3.40 (dt, J = 5.3, 4.2 Hz, 2H), 3.38 - 3.20 (m, 3H), 3.17 (dd, J = 11.5, 6.0 Hz, 1H), 2.99 - 2.89 (m, 2H), 2.74 (dt, J = 13.9, 4.8 Hz, 1H), 2.51 (dt, J = 13.9, 5.0 Hz, 1H). 13 C NMR (100 MHz, Chloroform-d) δ 171.33, 167.24, 136.54, 136.44, 134.03, 134.01, 133.97, 133.91, 133.27, 131.71, 131.35, 131.11, 130.93, 129.89, 129.80, 129.77, 129.70, 129.29, 129.24, 129.22, 129.19, 128.67, 127.40, 126.80, 126.30, 126.28, 126.25, 126.20, 125.23, 123.96, 123.26, 122.57, 121.88, 121.39, 119.68, 111.42, 108.14, 55.06, 53.62, 52.13, 39.59, 39.41, 31.48, 21.49.

[0146]

[0147] Example 3, Preparation of Hirsutine alkaloid derivatives modified with benzyl or heterocycle-fused benzyl groups containing different substituents

[0148] This reaction example uses compound II as starting material, hydrolysis is carried out under the condition of lithium hydroxide, pH is adjusted to about 3 with 2N dilute hydrochloric acid, then condensation with mono-Boc protected aromatic diamine or aliphatic diamine, followed by de-Boc, the obtained intermediate undergoes nucleophilic substitution reaction with bromobenzyl or heterocycle-fused bromobenzyl containing different substituents to obtain the Hirsutine alkaloid derivatives modified with benzyl or heterocycle-fused benzyl groups containing different substituents, the preparation route is shown in the following figure: Figure 3

[0149] The Hirsutine alkaloid derivatives modified with benzyl or heterocycle-fused benzyl groups containing different substituents described in this example have the following structure:

[0150]

[0151] In the formula, R is selected from one of the following groups: wherein R5 is o-NHCH2, m-NHCH2, p-NHCH2; or n = 1, 2, 3, 4, 5, R6 is H or CH3; or In the formula, R1 is substituted at any position on the benzene ring and is selected from one of the following groups: CH3, CH2CH3, F, Cl, Br, CN. In the formula, one of R2, R3, R4 is H, and the other two substituents can be the same or different, selected from H, CH3, OCH3, F, NO2, CF3, NH2, OH; or R4 is H, and R2 and R3 are one of the following groups forming a saturated or unsaturated five- or six-membered ring through C, O, N, S atoms:

[0152] 1. Preparation of N-(2-(benzylamino)phenyl)-1,4,6,7,12,12b-hexahydroindolo[2,3- a]quinoline-3-carboxamide (compound I-32)

[0153] ​In a 100 mL single necked flask, N-(2-aminophenyl)-l,4,6,7,12,12b- hexahydroindolo[2,3-a]quinoline-3-carboxamide (151 mg, 0.42 mmol, 1.5 eq) prepared by the same procedure as in step 1-3 of Example 2, benzyl bromide (48 mg, 0.28 mmol, 1.0 eq), anhydrous potassium carbonate (116 mg, 0.84 mmol, 3.0 eq) were added, followed by DMF 5 mL, stirred at room temperature for 4 h. After TLC test showed the reaction was complete, about 100 mL of water was added to the reaction, extracted with ethyl acetate for three times, washed with saturated NaCl once, and column chromatography (DCM:MeOH=25:1) to get 90 mg of white solid, with a yield of 72%. 1 H NMR (400 MHz, DMSO-d6) δ 10.54 (s, 1H), 9.45 (s, 1H), 7.56 - 7.50 (m, 2H), 7.39 - 7.20 (m, 8H), 7.16 - 7.07 (m, 2H), 7.03 - 6.92 (m, 2H), 6.22 (t, J = 5.3 Hz, 1H), 4.54 (dt, J = 5.3, 0.9 Hz, 2H), 4.49 (td, J = 4.9, 1.7 Hz, 1H), 3.97 (dq, J = 13.5, 1.0 Hz, 1H), 3.84 (dq, J = 13.6, 1.1 Hz, 1H), 3.30 - 3.17 (m, 2H), 2.99 - 2.87 (m, 2H), 2.73 (dt, J = 14.1, 4.9 Hz, 1H), 2.48 (dt, J = 14.1, 4.9 Hz, 1H). 13 C NMR (100 MHz, DMSO-d6) δ 169.35, 139.58, 138.10, 137.54, 136.40, 133.27, 130.06, 128.65, 127.97, 127.82, 126.79, 123.86, 123.85, 123.72, 121.98, 121.44, 120.39, 119.08, 115.91, 111.52, 108.04, 55.03, 53.48, 52.18, 49.61, 31.77, 21.63.

[0154]

[0155] 2. Preparation of N-(2-((3-methylbenzyl)amino)phenyl)-l,4,6,7,12,12b- hexahydroindolo[2,3-a]quinoline-3-carboxamide (Compound I-33)

[0156] Prepared in the same manner as for Example 3, 1, except substituting 3- methylbromobenzyl for the starting material, to give a white solid, 92 mg, 71% yield. 1 H NMR (400 MHz, DMSO-d6) δ 10.52 (s, 1H), 9.47 (s, 1H), 7.56 - 7.49 (m, 2H), 7.34 - 7.25 (m, 2H), 7.24 - 7.19 (m, 2H), 7.16 - 7.08 (m, 3H), 7.06 - 7.01 (m, 2H), 6.99 - 6.92 (m, 2H), 6.57 (t, J = 5.5 Hz, 1H), 4.60 (dt, J = 5.5, 0.9 Hz, 2H), 4.45 (td, J = 5.0, 1.8 Hz, 1H), 3.96 (dq, J = 13.7, 1.2 Hz, 1H), 3.80 (dq, J = 13.5, 1.0 Hz, 1H), 3.25 - 3.17 (m, 2H), 2.99 - 2.85 (m, 2H), 2.70 (dt, J = 14.1, 4.9 Hz, 1H), 2.54 (dt, J = 14.1, 4.9 Hz, 1H). 13 CNMR (100 MHz, DMSO-d6) δ 169.45, 138.85, 138.29, 138.22, 137.44, 136.50, 133.29, 130.01, 129.82, 129.02, 128.37, 126.87, 125.89, 123.91, 123.84, 123.70, 121.98, 121.44, 120.44, 119.28, 115.91, 111.62, 108.54, 55.18, 53.46, 52.11, 49.62, 31.75, 21.64, 19.63.

[0157]

[0158] 3. Preparation of N-(2-((3,4-dimethylbenzyl)amino)phenyl)-1,4,6,7,12,12b- hexahydroindolo[2,3-a]quinoline-3-carboxamide (Compound I-34)

[0159] Prepared in the same manner as for Example 3, 1, except substituting 3,4- dimethylbromobenzyl for the starting material, to give a white solid, 93 mg, 70% yield. 1H NMR (400 MHz, DMSO-d6) δ 10.64 (s, 1H), 9.55 (s, 1H), 7.56 - 7.51 (m, 2H), 7.34 - 7.29 (m, 1H), 7.24 (td, J=7.4, 2.7 Hz, 1H), 7.16 - 6.90 (m, 7H), 6.47 (t, J=5.4 Hz, 1H), 4.67 (dt, J=5.5, 0.9 Hz, 2H), 4.54 (td, J=5.1, 1.6 Hz, 1H), 3.95 (dq, J=13.5, 1.0 Hz, 1H), 3.88 (dq, J=13.6, 1.1 Hz, 1H), 3.26 - 3.17 (m, 2H), 2.99 - 2.90 (m, 2H), 2.63 (dt, J=14.1, 4.9 Hz, 1H), 2.55 (dt, J=14.1, 4.9 Hz, 1H), 2.29 (d, J=1.0 Hz, 2H). 13 C NMR (100 MHz, DMSO-d6) δ 169.49, 138.72, 137.44, 137.47, 136.76, 136.56, 136.50, 133.27, 130.16, 129.96, 129.43, 126.89, 126.61, 123.86, 123.85, 123.72, 122.06, 121.43, 120.35, 119.11, 115.96, 111.55, 108.09, 55.04, 53.55, 52.14, 49.13, 31.73, 21.57, 19.85, 19.59.

[0160]

[0161] 4. Preparation of N-(2-((4-nitrobenzyl)amino)phenyl)-1,4,6,7,12,12b- hexahydroindolo[2,3-a]quinoline-3-carboxamide (Compound 1-35)

[0162] Prepared in the same manner as described in step 1 of Example 3, except that the starting material was replaced by 4-nitrobenzyl bromide, to give 62 mg of white solid, yield 45%. 1H NMR (400 MHz, DMSO-d6) δ 10.58 (s, 1H), 9.67 (s, 1H), 8.13 - 8.08 (m, 2H), 7.60 - 7.48 (m, 4H), 7.34 - 7.28 (m, 2H), 7.26 (td, J=7.2, 2.6 Hz, 1H), 7.14 - 7.07 (m, 2H), 7.03 - 6.92 (m, 2H), 6.20 (t, J=5.3 Hz, 1H), 4.59 (dt, J=5.6, 0.9 Hz, 2H), 4.49 (td, J=4.9, 1.5 Hz, 1H), 3.97 (dq, J=13.3, 1.1 Hz, 1H), 3.84 (dq, J=13.5, 1.2 Hz, 1H), 3.35 - 3.17 (m, 2H), 2.99 - 2.87 (m, 2H), 2.73 (dt, J=14.0, 4.9 Hz, 1H), 2.48 (dt, J=14.2, 4.9 Hz, 1H). 13 CNMR (100 MHz, DMSO-d6) δ 169.35, 146.10, 144.96, 138.33, 137.54, 136.40, 133.27, 130.06, 129.31, 126.79, 123.86, 123.85, 123.72, 123.52, 121.98, 121.44, 120.39, 119.08, 115.91, 111.52, 108.04, 55.17, 53.68, 52.21, 48.89, 31.60, 21.88.

[0163]

[0164] 5. Preparation of N-(2-((4-fluorobenzyl)amino)phenyl)-l,4,6,7,12,12b- hexahydroindolo[2,3-a]quinoline-3-carboxamide (Compound I-36)

[0165] Prepared in the same manner as described in step 1 of Example 3, except that the starting material was replaced by 4-fluorobromobenzene to give 67 mg of white solid in 51% yield. 1H NMR (400 MHz, DMSO-d6) δ 10.43 (s, 1H), 9.37 (s, 1H), 7.54 - 7.48 (m, 2H), 7.39 - 7.29 (m, 4H), 7.21 (td, J=7.3, 2.4 Hz, 1H), 7.15 - 7.09 (m, 2H), 7.04 - 6.96 (m, 3H), 6.93 (dd, J=8.2, 1.4 Hz, 1H), 6.25 (t, J=5.5 Hz, 1H), 4.56 (dt, J=5.4, 0.9 Hz, 2H), 4.53 (td, J=4.9, 1.8 Hz, 1H), 3.94 (dq, J=13.7, 1.0 Hz, 1H), 3.81 (dq, J=13.4, 1.1 Hz, 1H), 3.35 (ddd, J=12.0, 6.0, 4.1 Hz, 1H), 3.19 (ddd, J=11.9, 6.3, 4.3 Hz, 1H), 2.95 - 2.85 (m, 2H), 2.71 (dt, J=14.1, 4.9 Hz, 1H), 2.54 (dt, J=14.1, 4.9 Hz, 1H). 13 C NMR (101 MHz, DMSO-d6) δ 169.37, 162.93, 160.91, 138.38, 137.52, 136.45, 135.93, 135.85, 133.28, 130.46, 130.39, 130.01, 126.83, 123.84, 123.86, 123.73, 121.94, 121.47, 120.44, 119.18, 115.94, 115.43, 115.25, 111.50, 108.01, 55.13, 53.45, 52.12, 48.41, 31.67, 21.73.

[0166]

[0167] 6. Preparation of N-(2-((3-methoxy-4-nitrobenzyl)amino)phenyl)-l,4,6,7,12,12b- hexahydroindolo[2,3-a]quinoline-3-carboxamide (Compound 1-37)

[0168] Prepared in the same manner as described in step 1 of Example 3, except substituting 3-methoxy-4-nitrobenzyl bromide as starting material, to give 67 mg of white solid in 49% yield. 1H NMR (400 MHz, DMSO-d6) δ 10.63 (s, 1H), 9.54 (s, 1H), 7.86 (d, J = 8.7 Hz, 1H), 7.56 - 7.48 (m, 2H), 7.38 (dd, J = 8.7, 1.7 Hz, 1H), 7.35 - 7.28 (m, 2H), 7.23 (td, J = 7.3, 2.8 Hz, 1H), 7.18 - 7.09 (m, 2H), 7.02 - 6.95 (m, 2H), 6.93 (dd, J = 8.1, 1.8 Hz, 1H), 6.71 (t, J = 5.5 Hz, 1H), 4.65 (dt, J = 5.7, 1.2 Hz, 2H), 4.46 (td, J = 5.2, 1.8 Hz, 1H), 3.95 (dq, J = 13.5, 1.3 Hz, 1H), 3.92 (s, 2H), 3.83 (dq, J = 13.7, 1.3 Hz, 1H), 3.27 - 3.17 (m, 2H), 2.97 - 2.88 (m, 2H), 2.72 (dt, J = 14.3, 4.9 Hz, 1H), 2.52 (dt, J = 14.0, 4.9 Hz, 1H). 13 CNMR (100 MHz, DMSO-d6) δ 169.38, 154.03, 144.14, 138.35, 137.56, 137.19, 136.42, 133.28, 130.07, 126.84, 125.79, 123.89, 123.83, 123.71, 121.94, 121.52, 121.44, 120.43, 119.12, 115.96, 113.43, 111.51, 108.06, 56.65, 55.08, 53.45, 52.13, 48.92, 31.53, 21.66.

[0169]

[0170] 7. Preparation of N-(3-((4-methoxybenzyl)amino)propyl)-l,4,6,7,12,12b- hexahydroindolo[2,3-a]quinoline-3-carboxamide (Compound I-38)

[0171] Prepared in the same manner as in Step 1 of Example 3, except that the starting material was replaced with 4-methoxybenzyl bromide and N-(3-aminopropyl)-l,4,6,7,12,12b- hexahydroindolo[2,3-a]quinoline-3-carboxamide prepared according to the same procedure as in Steps 1 to 3 of Example 2, to give a white solid 91 mg in 73% yield. 1H NMR (400 MHz, Chloroform-d) δ 8.42 (s, 1H), 7.51 (dd, J = 6.9, 2.4 Hz, 1H), 7.28 - 7.00 (m, 7H), 6.90 - 6.83 (m, 2H), 4.47 (td, J = 5.2, 1.9 Hz, 1H), 3.96 - 3.87 (m, 3H), 3.81 (s, 3H), 3.80 - 3.74 (m, 2H), 3.36 (ddd, J = 12.0, 6.4, 4.5 Hz, 1H), 3.20 - 3.14 (m, 3H), 2.98 - 2.88 (m, 2H), 2.85 (q, J = 5.4 Hz, 2H), 2.71 (dt, J = 14.2, 4.9 Hz, 1H), 2.55 (dt, J = 13.9, 5.1 Hz, 1H), 1.70 - 1.66 (m, 1H). 13 C NMR (100 MHz, Chloroform-d) δ 171.91, 159.14, 136.57, 136.45, 133.28, 132.79, 129.62, 126.83, 123.81, 121.51, 120.44, 119.18, 113.69, 111.52, 108.14, 55.15, 53.81, 53.66, 51.92, 49.93, 46.06, 39.09, 31.61, 28.40, 21.73.

[0172]

[0173] 8. Preparation of N-(3-((3-methyl-4-methoxybenzyl)amino)propyl)-1,4,6,7,12,12b- hexahydroindolo[2,3-a]quinoline-3-carboxamide (Compound 1-39)

[0174] Prepared in the same manner as in Step 1 of Example 3, except that the starting material was replaced with 3-methyl-4-methoxybenzyl bromide and N-(3-aminopropyl)-1,4,6,7,12,12b-hexahydroindolo[2,3-a]quinoline-3-carboxamide prepared in the same manner as in Steps 1-3 of Example 2, to give a white solid 95 mg, yield 74%. 1H NMR (400 MHz, Chloroform-d) δ 8.42 (s, 1H), 7.57 (dd, J = 7.1, 2.6 Hz, 1H), 7.33 (dd, J = 7.2, 2.3 Hz, 1H), 7.25 (td, J = 7.3, 2.4 Hz, 1H), 7.17 - 7.14 (m, 1H), 7.10 - 7.02 (m, 3H), 6.78 (d, J = 8.5 Hz, 1H), 4.64 (p, J = 5.5 Hz, 1H), 4.41 (td, J = 4.9, 1.5 Hz, 1H), 3.96 - 3.87 (m, 3H), 3.84 - 3.77 (m, 3H), 3.27 (ddd, J = 12.0, 6.2, 4.3 Hz, 1H), 3.21 - 3.11 (m, 3H), 2.99 - 2.91 (m, 2H), 2.86 (q, J = 5.4 Hz, 2H), 2.75 (dt, J = 14.2, 4.9 Hz, 1H), 2.50 (dt, J = 13.9, 4.8 Hz, 1H), 1.65 (d, J = 5.5 Hz, 2H). 13 C NMR (100 MHz, Chloroform-d) δ 171.81, 157.19, 136.60, 136.45, 133.67, 133.28, 130.18, 127.27, 126.96, 126.81, 123.87, 121.61, 120.44, 119.09, 111.54, 111.52, 108.14, 55.62, 54.07, 53.66, 52.12, 49.93, 46.06, 39.09, 31.61, 28.40, 21.43, 16.23.

[0175]

[0176] 9. Preparation of (1,4,6,7,12,12b-hexahydroindolo[2,3-a]quinolin-3-yl)(4-(3- methylbenzyl)piperazin-1-yl)methanone (Compound 1-40)

[0177] Prepared in the same manner as in Step 1 of Example 3, except that the starting material was replaced with 3-methylbenzyl bromide and (1,4,6,7,12,12b- hexahydroindolo[2,3-a]quinolin-3-yl)(piperazin-1-yl)methanone prepared according to the procedures of Steps 1-3 of Example 2, to give 96 mg of a white solid in 78% yield. 1H NMR (400 MHz, Chloroform-d) δ 8.55 (s, 1H), 7.59 (dd, J = 7.3, 2.6 Hz, 1H), 7.31 - 7.08 (m, 6H), 7.02 - 6.97 (m, 2H), 4.43 (td, J = 4.6, 1.9 Hz, 1H), 3.83 - 3.75 (m, 2H), 3.57 (t, J = 1.1 Hz, 2H), 3.50 - 3.43 (m, 4H), 3.38 - 3.21 (m, 2H), 2.96 - 2.89 (m, 2H), 2.88 - 2.78 (m, 4H), 2.65 (dt, J = 14.4, 5.3 Hz, 1H), 2.43 (dt, J = 14.3, 5.0 Hz, 1H). 13 C NMR (100 MHz, Chloroform-d) δ 169.46, 138.63, 137.77, 136.45, 136.12, 133.17, 129.47, 129.41, 128.49, 126.73, 126.13, 124.50, 123.85, 120.36, 119.03, 111.55, 108.06, 60.30, 55.26, 53.51, 52.16, 51.48, 44.78, 31.50, 21.68, 19.93.

[0178]

[0179] Example 4, Preparation of Hirsutine alkaloid derivatives modified with anilines or hetero-fused anilines containing different substituents

[0180] This reaction example uses compound II as the starting material, hydrolysis is carried out under the condition of lithium hydroxide, then the pH is adjusted to about 3 with 2N dilute hydrochloric acid, and then condensation reaction is carried out with anilines or hetero-fused anilines containing different substituents to obtain the Hirsutine alkaloid derivatives modified with anilines or hetero-fused anilines containing different substituents, and the preparation route is shown in the following figure: Figure 4

[0181] The Hirsutine alkaloid derivatives modified with anilines or hetero-fused anilines containing different substituents described in this example have the following structure:

[0182]

[0183] ​wherein R is NH. R1is substituted at any position on the phenyl ring and is selected from one of the following groups: CH3, CH2CH3, F, Cl, Br, CN. One of R2, R3, R4is H and the other two can be the same or different and are selected from H, CH3, OCH3, F, NO2, CF3, NH2, OH; or R4is H and R2and R3are one of the following groups forming a saturated or unsaturated five or six membered ring through a C, O, N, S atom:

[0184] 1. Preparation of N-(4-methoxyphenyl)-l,4,6,7,12,12b-hexahydroindolo[2,3- a]quinoline-3-carboxamide (Compound 1-41)

[0185] Prepared in the same manner as described in Example 2, steps 1-2, except that in step 2 one of the starting materials was replaced with 4-methoxyaniline to give 96 mg of white solid in 66% yield. 1 H NMR (400 MHz, Chloroform-d) δ 8.43 (d, J = 1.0 Hz, 2H), 7.58 (dd, J = 6.7, 2.3 Hz, 1H), 7.30 - 7.21 (m, 4H), 7.16 - 7.07 (m, 2H), 6.95 - 6.91 (m, 2H), 4.53 (td, J = 5.2, 1.9 Hz, 1H), 3.95 - 3.88 (m, 2H), 3.82 (s, 2H), 3.33 - 3.17 (m, 2H), 2.97 - 2.89 (m, 2H), 2.71 (dt, J = 13.8, 4.9 Hz, 1H), 2.51 (dt, J = 14.0, 4.7 Hz, 1H). 13 CNMR (100 MHz, Chloroform-d) δ 169.42, 157.25, 137.68, 136.40, 133.27, 132.98, 126.79, 123.86, 122.24, 121.41, 120.39, 119.08, 114.69, 111.52, 108.04, 55.35, 53.53, 52.05, 49.62, 31.43, 21.73.

[0186]

[0187] 2. Preparation of N-(4-methoxyphenyl)-l,4,6,7,12,12b-hexahydroindolo[2,3- a]quinoline-3-carboxamide (Compound 1-41)

[0188] Prepared by the same method as for example 2, steps 1-2, except that in step 2 one of the starting materials was replaced with 4-methylaniline to give a white solid 98 mg, 70% yield. 1 H NMR (400 MHz, Chloroform-d) δ 8.42 (s, 1H), 8.35 (s, 1H), 7.56 (dd, J = 6.6, 2.1 Hz, 1H), 7.45 - 7.41 (m, 2H), 7.30 - 7.27 (m, 1H), 7.25 (td, J = 7.5, 2.8 Hz, 1H), 7.16 - 7.07 (m, 4H), 4.45 (td, J = 4.7, 1.5 Hz, 1H), 3.99 - 3.87 (m, 2H), 3.31 - 3.19 (m, 2H), 2.96 - 2.88 (m, 2H), 2.71 (dt, J = 14.1, 4.9 Hz, 1H), 2.51 (dt, J = 14.1, 4.9 Hz, 1H), 2.37 (s, 2H). 13 C NMR (100 MHz, Chloroform-d) δ 169.51, 137.74, 137.50, 136.42, 133.29, 132.91, 129.57, 126.84, 123.83, 122.40, 120.43, 119.99, 119.09, 111.50, 108.04, 55.12, 53.45, 52.12, 31.69, 21.72, 19.93.

[0189]

[0190] 3. Preparation of N-(4-fluorophenyl)-l,4,6,7,12,12b-hexahydroindolo[2,3- a]quinoline-3-carboxamide (Compound I-43)

[0191] Prepared by the same method as for example 2, steps 1-2, except that in step 2 one of the starting materials was replaced with 4-fluoroaniline to give a white solid 100 mg, 71% yield. 1H NMR (400 MHz, Chloroform-d) δ 8.46 (s, 1H), 8.37 (s, 1H), 7.50 (dd, J = 7.0, 2.2 Hz, 1H), 7.49 - 7.45 (m, 2H), 7.31 (dd, J = 7.0, 1.9 Hz, 1H), 7.25 - 7.21 (m, 1H), 7.13 (td, J = 7.1, 1.8 Hz, 1H), 7.11 - 7.06 (m, 2H), 7.08 - 7.04 (m, 1H), 4.43 (td, J = 5.1, 1.8 Hz, 1H), 3.95 - 3.87 (m, 2H), 3.32 - 3.15 (m, 2H), 2.99 - 2.95 (m, 2H), 2.76 (dt, J = 14.2, 5.3 Hz, 1H), 2.55 (dt, J = 14.0, 4.7 Hz, 1H). 13 C NMR (100 MHz, Chloroform-d) δ 169.42, 159.40, 157.42, 137.68, 136.40, 136.21, 136.18, 133.27, 126.79, 123.86, 121.96, 121.89, 121.41, 120.39, 119.08, 115.89, 115.71, 111.52, 108.04, 55.03, 53.45, 52.18, 31.66, 21.78.

[0192]

[0193] 4. Preparation of N-(3-nitrophenyl)-l,4,6,7,12,12b-hexahydroindolo[2,3- a]quinoline-3-carboxamide (Compound I-44)

[0194] Prepared in the same manner as described in Example 2, Step 1-2, except that one of the starting materials in Step 2 was replaced with 3-nitroaniline, to give a white solid 79 mg, yield 52%. 1H NMR (400 MHz, Chloroform-d) δ 9.01 (s, 1H), 8.43 (s, 1H), 8.33 (t, J = 2.3 Hz, 1H), 8.21 (dt, J = 7.3, 2.0 Hz, 1H), 7.65 - 7.56 (m, 2H), 7.54 (dd, J = 6.9, 2.4 Hz, 1H), 7.29 (dd, J = 6.9, 2.1 Hz, 1H), 7.27 - 7.24 (m, 1H), 7.13 - 7.05 (m, 2H), 4.56 (td, J = 5.2, 1.8 Hz, 1H), 4.01 - 3.86 (m, 2H), 3.31 - 3.17 (m, 2H), 2.99 - 2.96 (m, 2H), 2.70 (dt, J = 14.1, 4.9 Hz, 1H), 2.52 (dt, J = 14.1, 4.9 Hz, 1H). 13 C NMR (100 MHz, Chloroform-d) δ 169.32, 148.82, 140.00, 137.68, 136.40, 133.27, 131.62, 126.96, 126.81, 123.86, 122.33, 120.43, 119.06, 117.29, 113.71, 111.52, 108.22, 55.18, 53.45, 52.02, 31.45, 21.81.

[0195]

[0196] Example 5, Anti-influenza virus activity of partial Hirsutine alkaloid derivatives

[0197] The partial Hirsutine derivatives synthesized above were selected in this example to evaluate the ability to resist influenza virus H1N1. The compound concentration was 30 μM, the cell strain was MDCK cells, the screening method was CPE inhibition experiment, and the evaluation standard was the inhibition rate of virus-induced cytopathic effect (CPE) or the protection rate of cells as the index. Ribavirin with broad-spectrum antiviral activity was selected as the positive control drug. The results are shown in Table 1. Most of the test compounds showed certain anti-influenza virus activity. Among them, Hirsutine derivatives I-1, I-2 and I-20 had 67.3%, 63.4% and 74% inhibition rates against the virus, respectively, which were slightly lower than the antiviral activity of the positive control drug ribavirin (77.9%) at a concentration of 25 μM.

[0198]

[0199] Table 1 Inhibition rate of Hirsutine derivatives on virus (H1N1)

[0200]

[0201]

[0202] Example 6, Cytotoxicity of Hirsutine alkaloid derivatives

[0203] We selected five compounds I-1, I-2, I-6, I-12, I-20 which had an inhibition rate of more than 50% at the initial screening concentration of 30 μM, and determined the cytotoxicity of the compounds on 293T, MDCK and A549 cells to evaluate the therapeutic index (SI), and the experimental results are shown in Tables 2, 3:

[0204] Table 2 Cytotoxicity of compounds on 293T human embryonic kidney cells, A549 cells

[0205]

[0206] Table 3 Cytotoxicity of compounds on MDCK cells

[0207]

[0208] From the above experimental results, it is found that the compounds have substantially no cytotoxicity (CC 50 > 500 μM) on MDCK cells; and the compound I-19 shows lower cytotoxicity on A549 cells and 239T cells. From the perspective of antiviral drugs, the Hirsutine alkaloid derivatives have potential research value.

Claims

1. Hirsutine alkaloid derivatives, characterized in that, having the structural formula shown below:

2. Use of the Hirsutine alkaloid derivative according to claim 1 for the preparation of a medicament for the prevention and / or treatment of a viral infection selected from the group consisting of an influenza A virus infection and an influenza B virus infection.

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