Indole fused hydrogenated cinnoline compound as well as preparation method and application thereof

Abstract

The invention discloses an indole fused hydrogenated cinnoline compound and a preparation method and application thereof, and belongs to the technical field of fluorescent materials and anti-tumor compounds, enaminone compounds and amine compounds are used as reaction substrates, and the indole fused hydrogenated cinnoline compound is obtained through a stork enamine reaction and an intramolecular acylation reaction in a high-polarity solvent. A series of indole fused hydrogenated cinnoline compounds containing different substituent groups are synthesized, the fluorescence wavelength of the compounds ranges from 430 nm to 700 nm, the compounds show certain inhibitory activity on lung cancer A549, breast cancer MDA-MB-231, colon cancer HCT116, liver cancer HepG2 and prostate cancer DU145, and more choices are provided for preparation of tumor drugs.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of fluorescent materials and antitumor compounds, and particularly relates to an indole-fused hydrogenated cinnoline compound and a preparation method and application thereof. BACKGROUND

[0002] Cinnoline compounds are six-membered heterocyclic compounds containing two nitrogen atoms. Their core skeletons are widely present in natural products and pharmaceutically active molecules. As important organic molecular skeletons, cinnoline compounds have wide applications in medicinal chemistry and material science. In the past few decades, researchers have developed various efficient synthetic methods. However, existing strategies often rely on expensive metal catalysts or harsh reaction conditions, which limits their application in industrial production. Therefore, it is still of important research value and practical significance to develop new methods for constructing cinnoline rings under mild conditions, with economic steps and environmental friendliness. SUMMARY

[0003] To solve the above technical problems, the present application provides an indole-fused hydrogenated cinnoline compound and a preparation method and application thereof.

[0004] To achieve the above-mentioned purpose, the present application provides the following technical solutions.

[0005] One of the technical solutions of the present application is as follows:

[0006] The present application provides an indole-fused hydrogenated cinnoline compound, the structural formula of which is as follows:

[0007]

[0008] wherein R is selected from H, halogen, methyl (Me), methoxy, cyano or nitro, R 1 is selected from hydroxyalkyl or alkyl.

[0009] The fluorescent wavelength range of the indole-fused hydrogenated cinnoline compound of the present application is between 430-700 nm, and it has good anti-proliferation effect on five cell strains of lung cancer A549, breast cancer MDA, colon cancer HCT116, liver cancer HepG2 and prostate cancer DU145, thereby providing a usable compound for the research and development of anticancer drugs.

[0010] Further, when the R is halogen, it includes monohalogen substitution and dihalogen substitution; the halogen is F, Cl or Br; the hydroxyalkyl is C2-C7 hydroxyalkyl (hydroxyethyl, hydroxypropyl, hydroxybutyl, hydroxyamyl, hydroxyhexyl, hydroxyheptyl); and the alkyl is n-butyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

[0011] Further, the indole-fused hydrogenated cinnoline compound is selected from the following compounds:

[0012] Further, the indole-fused hydrogenated cinnoline compound is selected from the following compounds:

[0013] The second technical solution of the present application is:

[0014] The present application also provides a preparation method of the above-mentioned indole-fused hydrocarbazole compound, which synthesizes the indole-fused hydrocarbazole compound by stork enamine reaction and intramolecular acylation reaction in a large polar solvent, taking enaminone compound and amine compound as reaction substrates,

[0015] The structural formula of the enaminone compound is wherein R is selected from H, halogen, methyl (Me), methoxy, cyano or nitro;

[0016] The structural formula of the amine compound is R 1 selected from hydroxyalkyl or alkyl.

[0017] Further, the molar ratio of the enaminone compound and the amine compound is 1:(1-10), more preferably 1:(1-5), and most preferably 1:5.

[0018] Further, the temperature of the stork enamine reaction and the intramolecular acylation reaction is 45-80℃, more preferably 60-80℃, and most preferably 80℃.

[0019] Further, the large polar solvent is selected from one of methanol, acetonitrile and chloroform, and preferably the large polar solvent is methanol.

[0020] The third technical solution of the present application is:

[0021] The present application also provides the application of the above-mentioned indole-fused hydrocarbazole compound in preparing fluorescent materials and / or semiconductor materials.

[0022] For example, the indole-fused hydrocarbazole compound of the present application can be prepared into organic light-emitting diode and / or fluorescent probe.

[0023] The fourth technical solution of the present application is:

[0024] The present application also provides the application of the above-mentioned indole-fused hydrocarbazole compound in preparing anti-tumor drugs.

[0025] Further, the anti-tumor drugs include lung cancer prevention and / or treatment drugs, breast cancer prevention and / or treatment drugs, liver cancer prevention and / or treatment drugs, and prostate cancer prevention and / or treatment drugs.

[0026] Compared with the prior art, the present application has the following advantages and technical effects:

[0027] The preparation method of the indole-fused hydrocarbazole compound is simple, the reaction condition is mild, the yield and purity of the indole-fused hydrocarbazole compound are relatively high, and the indole-fused hydrocarbazole compound can be prepared on a large scale.

[0028] The maximum fluorescence emission wavelength of the indole-fused hydrocarbazole compound is affected by the electronic effect of the substituent group. When the benzene ring has an electron-withdrawing group (-F, -Cl, -Br, -CN), the excitation wavelength and the emission wavelength of the product are slightly blue-shifted. When the benzene ring has an electron-donating group (-Me, -OMe), the excitation wavelength and the emission wavelength of the product are red-shifted. Moreover, the compound with the electron-donating group has weaker fluorescence and a larger stoke shift. The excitation wavelength and the emission wavelength of the product are slightly red-shifted by extending the carbon chain. After the chain is changed into a ring structure, the excitation wavelength and the emission wavelength of the compound are also slightly red-shifted. In general, the fluorescence wavelength range of the indole-fused hydrocarbazole compound is between 430-700 nm, the quantum yield is in the range of 15.3%-72.9%, and the indole-fused hydrocarbazole compound has good fluorescence performance. Simple modification of the structure of the indole-fused hydrocarbazole compound can change its fluorescence characteristics, and the indole-fused hydrocarbazole compound is expected to be applied in fluorescent materials and the like.

[0029] The indole-fused hydrocarbazole compound has a certain inhibitory effect on lung cancer A549, breast cancer MDA-MB-231, colon cancer HCT116, liver cancer HepG2 and prostate cancer DU145. Overall, the inhibitory effect of the compound on colon cancer HCT116 is stronger than that on the other four cancer cells, and the compound has a certain cell selectivity. From the analysis of the substituents of the compound, the four compounds with 4-position bromine substitution, 7-position bromine substitution, 4,6-position double chloro substitution and 4,7-position double chloro substitution have good proliferation inhibition effect on the five cancer cells, and the other compounds have not very strong cytotoxicity on the cancer cells, which indicates that the four compounds have a certain universality for cancer cells, and also confirms that the skeleton has a certain activity in anti-tumor and has a certain structure-activity relationship. BRIEF DESCRIPTION OF DRAWINGS

[0030] The accompanying drawings, which form a part of the present application, are used to provide further understanding of the present application, and serve as an explanation of the illustrative embodiments of the present application, and do not constitute an improper limitation of the present application. In the drawings:

[0031] Figure 1 is a structural formula and absolute configuration of compound 3aa;

[0032] Figure 2 is a synthetic mechanism route map of compound 3aa;

[0033] Figure 3 is the fluorescence spectrum of some compounds in 10 -5Fluorescence spectra of M in methanol solution;

[0034] Figure 4 Normalized fluorescence emission spectra of some compounds;

[0035] Figure 5 Fluorescence excitation and emission spectra of standards in methanol (10 -5 M). DETAILED DESCRIPTION

[0036] Various exemplary embodiments of the present application will now be described in detail, which should be considered to be illustrative of certain aspects, features and embodiments of the present application, but not a limitation thereof.

[0037] It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the present application. In addition, where a range of values is provided, it is understood that each intervening value, to the upper and lower limit of the range is also specifically disclosed. Each smaller range between any stated value or intervening value in the stated range and any other stated or intervening value in that stated range is encompassed. The upper and lower limits of these smaller ranges can independently be included or excluded in the range.

[0038] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. Although preferred methods and materials are described, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present application. All documents mentioned herein are incorporated by reference to disclose and describe in detail the methods and / or materials which are related to the present application. In the case of conflict between the present specification and any document incorporated herein by reference, the present specification will control.

[0039] Many modifications and variations of this application can be made in the light of the above teachings without departing from the spirit and scope thereof, and it is to be understood that all such modifications and variations warrant the patentable subject matter under the patent laws. Other embodiments of the application will be apparent to those skilled in the art from consideration of the specification and practice of the application disclosed herein. The specification and examples given are exemplary only.

[0040] As used herein, the terms "comprise", "comprising", "include", "including", "have", "having" and the like are open-ended and do not exclude additional elements or steps.

[0041] Unless otherwise specified, the room temperature in the present application is 25±2°C.

[0042] Each raw material used in the embodiments of the present application is commercially available.

[0043] The present application uses indole enamine ketone 1 and amino compound 2 as substrates, and synthesizes indole-fused hydrocinnoline compounds by site activation-intramolecular acylation reaction in a large polar solvent through Stork reaction, and the reaction route is as follows:

[0044]

[0045] wherein R is selected from H, halogen, methyl (Me), methoxy, cyano or nitro; R 1 selected from hydroxyalkyl or alkyl, and when the substituent R is halogen, it includes monohalogen substitution and dihalogen substitution.

[0046] In the embodiment of the present application, in the preparation method of the indole-fused hydrocinnoline compound, the reaction process is monitored by TLC thin layer chromatography, and the developing agent used is petroleum ether and ethyl acetate, and the volume ratio is petroleum ether: ethyl acetate = 1:1, and when the raw material point disappears, it is considered that the reaction is complete, and the reaction is complete at a temperature of 45-80 DEG C, and the time is 2-24 h. After the reaction is complete, stop the reaction, add an appropriate amount of silica gel, and dry the solvent by using a rotary evaporator; then the reaction system is separated by column chromatography, and the mobile phase used in the separation process is petroleum ether and ethyl acetate, and the volume ratio is petroleum ether: ethyl acetate = (1-2):1, and the whole separation process is monitored by TLC thin layer chromatography, so as to obtain pure separation product, and the separation solvent of the product section is dried by using a rotary evaporator to obtain pure product.

[0047] The lung cancer A549, breast cancer MDA-MB-231, colon cancer HCT116, liver cancer HepG2 and prostate cancer DU145 cell strains used in the embodiment of the present application are purchased from Hunan Fenghui Biological Technology Co., Ltd.

[0048] In the embodiment of the present application, the maximum emission wavelength and excitation wavelength of the indole-fused hydrocinnoline compound are determined by fluorescence spectrum determination, the relative fluorescence quantum yield of each compound is obtained by reference method, it is found that the compound has good fluorescence performance, and it is proved that it has broad prospects in the field of preparing fluorescent materials.

[0049] It should be pointed out that the parts not described in detail in the present application are conventional operation means in the art, and are not the focus of the present application.

[0050] The technical solutions of the present application are further described below through examples.

[0051] Example 1

[0052] The stork reaction was used to synthesize compound 3aa by site activation-intramolecular acylation reaction in different solvents with compound 1a and compound 2a as reaction substrates. The specific steps were as follows: compound 1a was taken into a 10 mL reaction bottle, 0.5 mL of the corresponding reaction solvent was added, and then the reaction bottle was placed in a magnetic stirrer for stirring and dissolution. Then, compound 2a was added, and then the reaction was carried out. The reaction process was monitored by TLC. The developing agent used was petroleum ether and ethyl acetate, and the volume ratio was petroleum ether: ethyl acetate = 1:1. When the starting material point disappeared, the reaction was complete, and the reaction was stopped. Silica gel was added, and the solvent was dried by rotary evaporation. Then, the reaction system was separated by column chromatography. The mobile phase used in the separation process was petroleum ether and ethyl acetate, and the volume ratio was petroleum ether: ethyl acetate = 2:1. The whole separation process was monitored by TLC to obtain pure separation product. The solvent type, molar ratio of compound 1a and compound 2a, reaction temperature and time are shown in Table 1, and the reaction route is as follows:

[0053]

[0054] Table 1

[0055]

[0056]

[0057] Note: [b] The yield is the separation yield of product 3aa after purification.[c] Reaction for 24 h at the specified temperature. [d] At the specified feeding amount ratio

[0058] The reaction was followed to completion.

[0059] From Table 1, it can be seen that indole enamine ketone 1a and ethanolamine 2a were used as starting reaction substrates. The first to eleventh groups were screened for reaction solvents when the feeding amount ratio of 1a:2a was 1:1 (0.05 mmol / 0.05 mmol), the reaction temperature was 28°C, and the reaction solvent was 0.5 mL. The screened solvents included small polar toluene, dichloroethane, dichloromethane, chloroform and tert-butanol, medium polar diethyl ether, ethyl acetate, tetrahydrofuran and dioxane, and large polar methanol and acetonitrile. It can be seen that the reaction yield was very low (7%-8%) when dichloroethane, toluene and tert-butanol were used as solvents, and the reaction yield was improved (21%-22%) when tetrahydrofuran and diethyl ether were used as reaction solvents. In summary, when the reaction occurred in large polar methanol (group 4), the yield was better (33%), so methanol was determined as the optimal solvent for the reaction.

[0060] To further improve the yield of the reaction, the temperature was screened. When the temperature was raised to 45°C, the reaction was accelerated (Group 12); when the temperature continued to rise to 60°C, the yield was slightly improved (Group 13); and when the temperature was raised to 80°C, the yield was significantly improved, and the yield was increased to 72% in the same time (Group 14). Next, the feeding ratio was explored (Groups 15-17), and increasing the amount of substrate 2a could accelerate the reaction rate and improve the reaction yield. When the feeding amount was 1:3 (molar ratio, the same below), the yield was increased to 84%; when the feeding amount was continuously increased, the template reaction could be completely reacted, and the yield reached 99% (Groups 16-17). It is worth noting that when the feeding amount was 1:10, the reaction was only 3h faster than the reaction with a feeding amount of 1:5. Combining the concepts of environmental protection, economy and sustainable development, reducing unnecessary loss and improving raw material utilization, the molar ratio of 1:5 was finally selected as the best feeding ratio for the reaction.

[0061] Through a series of condition screening of the reaction, the Group 16 was finally determined as the optimal condition of the reaction, i.e. using indole-based enaminone compound 1a and ethanolamine 2a as the starting reaction substrates, the molar ratio of indole-based enaminone 1a:ethanolamine 2a was 1:5 (0.05mmol / 0.25mmol), 0.5mL of methanol was used as the solvent, and the reaction was carried out at 80°C, and this reaction condition was used for the subsequent synthesis of indole-fused hydrocarbazole compounds.

[0062] Example 2

[0063] Based on the optimal condition of Group 16 in Example 1, using compound 1 (0.2mmol, 1.0eq) and compound 2a (1.0mmol, 5.0eq) as the substrates, i.e. the molar ratio of compound 1 and compound 2a was 1:5, the stork reaction was carried out in methanol (2.0mL) to synthesize indole-fused hydrocarbazole compound 3 by site activation-intramolecular acylation reaction, and the synthetic route was as follows:

[0064]

[0065] wherein R is F, Cl, Br, methyl, methoxy, cyano or nitro; the reaction time, reaction product and yield are shown in Table 2.

[0066] Table 2

[0067]

[0068]

[0069] Note: The yield is the isolated yield of the main product 3 (including 3ba-3ua) after purification.

[0070] As shown in Table 2, the reaction showed good functional group tolerance, and a series of indole-fused hydrocinnol compounds 3aa-3ua were obtained with excellent yields (48%-94%).

[0071] The present application is substituted with different electronic substituents of different properties at different positions of the indole benzene ring. When different halogen atoms (-F, -Cl, -Br) are substituted at the 4 position of the indole benzene ring, the overall impact on the reaction is not great (3ba, 3ca, 3da), and the yield is 89%, 91%, and 78%, respectively. When an electron-donating group (-Me) is introduced at the 4 position, the reaction time is prolonged, and it takes 13 h to complete the reaction, but the yield remains at 90%. Different electron-withdrawing groups (-F, -Cl, -Br, -CN) and electron-donating groups (-CH3, -OCH3) are then introduced at the 5 position of the indole. When halogen atoms (-F, -Cl, -Br) are introduced, the yield remains good (70%-94%); when the electron-withdrawing group -CN is introduced, the reaction rate may be accelerated due to its passivation effect on the benzene ring of the indole, and the reaction can be completed in 4 h, with a good yield (91%). When the substituent is -Me, 3ia has an excellent yield (90%), but the reaction rate is slowed down (13 h); and when the 5 position is substituted with the electron-donating group -OMe, the yield decreases (62%). Different electronic groups are then introduced at the 6 position (3la-3na), and the yield is well maintained (77%-94%). When the 7 position of the indole benzene ring is substituted (-Cl, -Br, -Me), the steric hindrance at the 7 position may be large, which limits the formation of intermediates during the reaction, so the reaction is relatively slow, and the yield decreases, with yields of 55%, 48%, and 62%, respectively. However, when the electron-withdrawing group -NO2 is introduced at the 7 position, the steric hindrance does not affect the reaction rate, but rather, -NO2 may lower the electron cloud density on the benzene ring through an inductive effect, and some of the electrons on the pyrrole ring delocalize to the benzene ring, thereby increasing the reactivity of the carbonyl carbon at the 2 position of the indole, so the reaction rate is accelerated, and the reaction is completed in 3 h, with a good yield (85%). Finally, halogen atoms are double-substituted at different positions of the indole benzene ring (3sa, 3ta, 3ua). Similarly, considering the increased steric hindrance that may result from double substitution, the yield of such substitution products is affected. When double -Cl substitution occurs, the yield of 4, 6 double-substituted compounds (3sa) is 90%, the yield of 4, 7 double-substituted compounds (3ua) is 89%, and the yield of 5, 7 double-substituted compounds is only 72%. When the 7 position of the indole is replaced by an N atom to form a 7-N heteroindole, the reaction time is shortened, and the yield is increased, which may be due to the high electronegativity of the N atom, which increases the electron cloud density around it, and reduces the electron cloud density of other parts of the indole ring, thereby increasing the nucleophilic reactivity.

[0072] When the pyrrole ring is replaced by the indole ring, the product obtained is (3wa), the specific reaction process is as follows: pyrrole-based enamine ketone (0.2 mmol, 1.0 eq) and compound 2 (1.0 mmol, 5.0 eq) are used as substrates, and the stork reaction is carried out by site activation-intramolecular acylation in methanol (2.0 mL) for 11 h, and the reaction is complete. The yield of the reaction is significantly reduced, only 11%, which may be due to the fact that, compared with indole, the pyrrole ring only has N lone pair electrons conjugated with the ring, and the lack of a benzene ring conjugation makes the electron density of the pyrrole ring larger than that of indole, and the nucleophilic reactivity is low, so the yield is reduced.

[0073] After substitution of different substituents with different properties at different positions of the indole benzene ring, it is found that different properties of the substituted electron groups have a certain influence on the reaction results, but basically the yield is good. In summary, the steric hindrance has a relatively greater influence on the reaction yield, and the yield of the 7-substituted product is reduced; secondly, the introduction of an electron-donating group-Me will prolong the reaction time, but the yield remains good; thirdly, when an electron-withdrawing group-CN and-NO2 is introduced, its inductive effect dominates, which can accelerate the reaction and also obtain an excellent yield.

[0074] Example 3

[0075] On the basis of the optimal conditions of group 16 in example 1, compound 1a (0.2 mmol, 1.0 eq) and compound 2 (1.0 mmol, 5.0 eq) are used as substrates, i.e. the molar ratio of compound 1a and compound 2 is 1:5, and the stork reaction is carried out by site activation-intramolecular acylation in methanol (2.0 mL) to synthesize indole-fused hydrogenated cinnoline compounds 3, and the synthesis route is as follows:

[0076]

[0077] wherein R is C2-C7 hydroxyalkyl, n-butyl, t-butyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl; the reaction time, reaction product and yield are shown in Table 3.

[0078] Table 3

[0079]

[0080]

[0081] Note: The yield is the separation yield of the main product 3 (including 3ab-3al) after purification.

[0082] Table 3 shows that, although the yield of the target compounds (3ab-3af) decreases somewhat with increasing carbon atom number, it remains relatively high (77%-88%). When the substrate is replaced with tert-butylamine, the reaction slows and the yield decreases, indicating that increased steric hindrance worsens the reaction. The present invention also replaces the chain amine with a cyclic amine, and the yield of the compounds (3ai-3al) (53%-87%) increases with increasing ring length. The amino group acts as a nucleophile to attack the protonated carbonyl carbon, forming a tetrahedral intermediate. Since the smaller the ring length of the cyclic amine, the greater the tension, the electrons become more crowded. The rigid conformation restricts adjacent sites, reducing the reactivity of the carbonyl's α-C. By expanding the substrate of amine compound 2, it was found that when ethanolamine is replaced with a long-chain amine or cyclic amine structure, the yield of the other compounds remains high, except for the tert-butylamine compound, which has a decreased yield, demonstrating the good functional group tolerance of the reaction.

[0083] Example 4 Amplification reaction

[0084] In order to investigate the potential of the reaction of the present invention in industrial production, compound 1a ( 835 mg, 2.8 mmol) was placed in a 100 mL round-bottom flask, 28 mL of methanol was added to dissolve, and then compound 2a ( 840 μL, 14.0 mmol), and reacted at 80°C, followed by TLC until the reaction was complete, and the solvent was dried and purified by column chromatography to obtain product 3aa ( 809 mg, 2.74 mmol), with a yield of 98%. After the scale-up reaction, the product can still maintain an excellent yield, proving that the reaction has the potential for industrial production.

[0085] In order to determine the configuration of the product, single crystals of compound 3aa were grown and the absolute configuration of compound 3aa was obtained after X-Ray crystal diffraction analysis ( Figure 1 ). Based on the absolute configuration of compound 3aa, the absolute configurations of other compounds are inferred. The synthetic mechanism of compound 3aa is shown in the following figure. Figure 2 As shown, the reaction mechanism is as follows: the lone pair of electrons from the primary amine 2 first attacks the carbonyl group of the enaminoketone 1, undergoing nucleophilic addition to yield the hemiketalamine intermediate A. This then eliminates a water molecule to yield the enamine B, which tautomerizes to form an imine. The resulting intermediate C is a more stable secondary ketimine. Intermediate C possesses both imine and enamine structures, with both the nitrogen atom and the β-carbon atom of the enamine being nucleophilic sites. The β-carbon atom of the enamine (which, due to the inductive effect of the electron-withdrawing imine functional group, has a stronger nucleophilic ability) nucleophilically attacks the ester carbonyl group at the 2-position of the indole, leading to intramolecular cyclization to yield compound 3aa.

[0086] The crystal structure data of the compound synthesized above are as follows:

[0087] Compound 3aa, yellow solid, 57.7 mg, 98% yield, mp 176.2-177.7 °C. 1 H NMR (400 MHz, DMSO-d6) δ 13.30 (s, 1H), 7.88 (d, J = 8.2 Hz, 1H), 7.76 (d, J = 8.0 Hz, 1H), 7.35 - 7.28 (m, 1H), 7.25 - 7.18 (m, 1H), 7.00 (s, 1H), 5.18 (t, J = 4.9 Hz, 1H), 3.68 (q, J = 4.9 Hz, 2H), 3.59 (q, J = 5.2 Hz, 2H), 2.83 (t, J = 6.1 Hz, 2H), 2.80 - 2.72 (m, 2H), 1.94 (p, J = 6.2 Hz, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 173.83, 172.68, 151.57, 133.13, 130.98, 125.36, 123.30, 122.17, 122.04, 111.61, 99.04, 95.69, 59.66, 46.46, 29.97, 27.13, 20.83. HRMS (ESI) m / z calcd for C 17 H 18 N3O2[M+H] + = 296.1399, found = 296.1403.

[0088] Compound 3ba, yellow solid, 56.6 mg, 89% yield, mp 214.6-215.2 °C. 1 H NMR (400 MHz, DMSO-d6) δ 13.30 (s, 1H), 7.88 (d, J = 8.2 Hz, 1H), 7.76 (d, J = 8.0 Hz, 1H), 7.35 - 7.28 (m, 1H), 7.25 - 7.18 (m, 1H), 7.00 (s, 1H), 5.18 (t, J = 4.9 Hz, 1H), 3.68 (q, J = 4.9 Hz, 2H), 3.59 (q, J = 5.2 Hz, 2H), 2.83 (t, J = 6.1 Hz, 2H), 2.80 - 2.72 (m, 2H), 1.94 (p, J = 6.2 Hz, 2H). 13C NMR (101 MHz, DMSO-d6) δ 173.42, 173.08, 156.50 (d, J = 247.0 Hz), 152.56, 135.37 (d, J = 11.0 Hz), 131.44, 123.64 (d, J = 7.6 Hz), 114.87 (d, J = 23.5 Hz), 108.24 (d, J = 3.5 Hz), 106.10 (d, J = 18.2 Hz), 99.23, 90.94, 59.59, 46.56, 29.92, 27.17, 20.72. 19 F NMR (376 MHz, DMSO-d6) δ -121.69 (dd, J = 10.9, 5.4 Hz). HRMS (ESI) m / z calcd for C 17 H 17 N3O2F [M + H] + = 314.1305, found = 314.1309.

[0089] Compound 3ca, yellow solid, 60.0 mg, 91% yield, mp 237.6-238.7 °C. 1 H NMR (400 MHz, DMSO-d6) δ 13.31 (s, 1H), 7.86 (s, 1H), 7.31 (s, 2H), 6.96 (s, 1H), 5.18 (s, 1H), 3.70 (s, 2H), 3.63 (s, 2H), 2.87 (s, 2H), 2.78 (s, 2H), 1.96 (s, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 173.42, 173.08, 156.50 (d, J = 247.0 Hz), 152.56, 135.37 (d, J = 11.0 Hz), 131.44, 123.64 (d, J = 7.6 Hz), 114.87 (d, J = 23.5 Hz), 108.24 (d, J = 3.5 Hz), 106.10 (d, J = 18.2 Hz), 99.23, 90.94, 59.59, 46.56, 29.92, 27.17, 20.72. 17 H 17 N3O2Cl [M + H] + = 330.1009, found = 330.1013.

[0090] Compound 3da, yellow solid, 59.2 mg, 78% yield, mp 231.8-232.6 °C. 1H NMR (400 MHz, DMSO-d6) δ 13.30 (s, 1H), 7.90 (d, J = 8.3 Hz, 1H), 7.46 (d, J = 7.3 Hz, 1H), 7.23 (t, J = 7.8 Hz, 1H), 6.88 (s, 1H), 5.18 (t, J = 4.8 Hz, 1H), 3.70 (q, J = 4.9 Hz, 2H), 3.61 (q, J = 5.3 Hz, 2H), 2.85 (t, J = 5.3 Hz, 2H), 2.80-2.72 (m, 2H), 1.95 (p, J = 6.2 Hz, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 173.24, 173.14, 152.71, 133.34, 131.73, 125.74, 124.71, 124.09, 115.22, 111.34, 99.31, 94.86, 59.60, 46.60, 29.92, 27.17, 20.70. HRMS (ESI) m / z calcd for C 17 H 17 N3O2Br [M+H] + = 374.0504, found = 374.0508.

[0091] Compound 3ea, yellow solid, 55.9 mg, 90% yield, mp 199.5-201.0 °C. 1 H NMR (400 MHz, DMSO-d6) δ 13.30 (s, 1H), 7.69 (d, J = 8.3 Hz, 1H), 7.20 (t, J = 7.7 Hz, 1H), 7.00 (d, J = 6.2 Hz, 2H), 5.15 (s, 1H), 3.68 (d, J = 5.2 Hz, 2H), 3.61 (q, J = 4.9, 4.3 Hz, 2H), 2.86 (t, J = 6.2 Hz, 2H), 2.76 (t, J = 6.2 Hz, 2H), 2.56 (s, 3H), 1.95 (p, J = 6.1 Hz, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 173.87, 172.66, 151.61, 132.98, 131.09, 130.53, 125.56, 123.52, 121.79, 109.22, 99.06, 94.38, 59.72, 46.46, 30.01, 27.15, 20.85, 18.79. HRMS (ESI) m / z calcd for C 18 H 20 N3O2 [M+H] += 310.1556, found = 310.1556.

[0092] Compound 3fa, yellow solid, 58.7 mg, 94% yield, mp 260.1-262.4 °C. 1 H NMR (400 MHz, DMSO-d6) δ 13.34 (s, 1H), 7.88 (dd, J = 8.8, 4.6 Hz, 1H), 7.50 (dd, J = 9.9, 2.5 Hz, 1H), 7.16 (td, J = 9.2, 2.6 Hz, 1H), 6.97 (s, 1H), 5.17 (s, 1H), 3.69 (s, 2H), 3.65-3.58 (m, 2H), 2.88 (t, J = 5.5 Hz, 2H), 2.81-2.72 (m, 2H), 1.95 (p, J = 6.3 Hz, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 173.34, 173.05, 158.63 (d, J = 235.1 Hz), 151.98, 132.30, 129.99, 125.36 (d, J = 11.0 Hz), 113.05 (d, J = 10.0 Hz), 112.26 (d, J = 27.2 Hz), 106.00 (d, J = 23.4 Hz), 99.14, 95.48 (d, J = 5.4 Hz), 59.61, 46.54, 29.90, 27.18, 20.80. 19 F NMR (376 MHz, DMSO-d6) δ -121.19 (td, J = 9.8, 4.5 Hz). HRMS (ESI) m / z calcd for C 17 H 17 N3O2F [M+H] + = 314.1305, found = 314.1308.

[0093] Compound 3ga, yellow solid, 60.0 mg, 91% yield, mp 252.4-253.8 °C. 1 H NMR (400 MHz, DMSO-d6) δ 13.31 (s, 1H), 7.84 (d, J = 21.0 Hz, 2H), 7.29 (s, 1H), 6.96 (s, 1H), 5.17 (s, 1H), 3.66 (d, J = 17.9 Hz, 4H), 2.89 (s, 2H), 2.77 (s, 2H), 1.96 (s, 2H). 13C NMR (101 MHz, DMSO-d6) δ 173.44, 173.15, 152.33, 132.14, 131.45, 126.51, 126.12, 123.38, 121.13, 113.30, 99.22, 95.18, 59.60, 46.58, 29.94, 27.21, 20.76. HRMS (ESI) m / z calcd for C 17 H 17 N3O2Cl [M+H] + = 330.1009, found = 330.1011.

[0094] Compound 3ha, yellow solid, 52.8 mg, 70% yield, mp 235.9-237.4 °C. 1 H NMR (400 MHz, DMSO-d6) δ 13.31 (s, 1H), 7.97 (s, 1H), 7.83 (d, J = 8.7 Hz, 1H), 7.40 (d, J = 8.3 Hz, 1H), 6.96 (s, 1H), 5.17 (s, 1H), 3.66 (dt, J = 16.8, 5.3 Hz, 4H), 2.90 (t, J = 6.2 Hz, 2H), 2.77 (t, J = 6.2 Hz, 2H), 1.96 (p, J = 6.3 Hz, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 173.44, 173.15, 152.33, 132.14, 131.45, 126.51, 126.12, 123.38, 121.13, 113.30, 99.22, 95.18, 59.60, 46.58, 29.94, 27.21, 20.76. HRMS (ESI) m / z calcd for C 17 H 17 N3O2Br [M+H] + = 374.0504, found = 374.0504.

[0095] Compound 3ia, yellow solid, 57.4 mg, 93% yield, mp 225.1-226.2 °C. 1H NMR (400 MHz, DMSO-d6) δ 13.24 (s, 1H), 7.69 (d, J = 8.5 Hz, 1H), 7.43 (s, 1H), 7.07 (d, J = 8.5 Hz, 1H), 6.82 (s, 1H), 5.10 (s, 1H), 3.61 (t, J = 5.1 Hz, 2H), 3.53 (q, J = 5.4 Hz, 2H), 2.79 (t, J = 6.2 Hz, 2H), 2.68 (t, J = 6.2 Hz, 2H), 2.35 (s, 3H), 1.87 (p, J = 6.1 Hz, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 173.80, 172.67, 151.30, 131.74, 130.94, 130.85, 125.59, 125.35, 121.25, 111.34, 99.02, 95.12, 59.68, 46.44, 29.96, 27.15, 21.78, 20.86. HRMS (ESI) m / z calcd for C 18 H 20 N3O2[M+H] + = 310.1556, found = 310.1557.

[0096] Compound 3ja, yellow solid, 40.1 mg, 62% yield, mp 214.5-215.7 °C. 1 H NMR (400 MHz, DMSO-d6) δ 13.35 (s, 1H), 7.76 (d, J = 9.0 Hz, 1H), 7.18 (s, 1H), 6.95 (d, J = 8.5 Hz, 1H), 6.89 (s, 1H), 5.15 (s, 1H), 3.80 (s, 3H), 3.68 (s, 2H), 3.61 (t, J = 5.3 Hz, 2H), 2.85 (t, J = 6.1 Hz, 2H), 2.75 (t, J = 6.1 Hz, 2H), 1.98 - 1.89 (m, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 173.45, 172.74, 155.50, 151.29, 131.14, 128.69, 125.81, 115.06, 112.52, 101.83, 99.00, 95.19, 59.67, 55.70, 46.45, 29.91, 27.13, 20.88. HRMS (ESI) m / z calcd for C 18 H 20 N3O3[M+H] + = 326.1505, found = 326.1509.

[0097] Compound 3ka, yellow solid, 58.2 mg, 91% yield, mp 267.6-268.2 °C. 1 H NMR (400 MHz, DMSO-d6) δ 13.27 (s, 1H), 8.35 (s, 1H), 8.01 (d, J = 8.6 Hz, 1H), 7.59 (d, J = 8.6 Hz, 1H), 7.12 (s, 1H), 5.19 (s, 1H), 3.75 - 3.56 (m, 4H), 2.94 - 2.86 (m, 2H), 2.83 - 2.75 (m, 2H), 2.01 - 1.91 (m, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 173.37, 173.31, 153.45, 133.83, 133.05, 128.61, 124.92, 124.68, 120.64, 113.02, 104.15, 99.31, 96.58, 59.54, 46.65, 29.97, 27.25, 20.64. HRMS (ESI) m / z calcd for C 18 H 17 N4O2[M+H] + = 321.1352, found = 321.1353.

[0098] Compound 3la, yellow solid, 59.4 mg, 94% yield, mp 204.9-206.5 °C. 1 H NMR (400 MHz, DMSO-d6) δ 13.25 (s, 1H), 7.79 (dd, J = 8.8, 5.4 Hz, 1H), 7.57 (dd, J = 9.7, 2.3 Hz, 1H), 7.10 (td, J = 9.3, 2.4 Hz, 1H), 7.02 (s, 1H), 5.17 (t, J = 4.5 Hz, 1H), 3.69 (q, J = 4.5 Hz, 2H), 3.60 (q, J = 5.2 Hz, 2H), 2.85 (t, J = 6.1 Hz, 2H), 2.78 - 2.72 (m, 2H), 1.94 (p, J = 6.3 Hz, 2H). 13CNMR (101 MHz, DMSO-d6) δ 173.71, 172.71, 159.94 (d, J = 238.9 Hz), 152.12, 132.72 (d, J = 12.5 Hz), 131.93 (d, J = 3.6 Hz), 123.92 (d, J = 10.1 Hz), 122.17, 111.43 (d, J = 25.4 Hz), 98.85, 97.10 (d, J = 26.3 Hz), 96.05, 59.64, 46.47, 29.95, 27.14, 20.75. 19 F NMR (376 MHz, DMSO-d6) δ -117.68 (td, J = 9.7, 5.4 Hz). HRMS (ESI) m / z calcd for C 17 H 17 N3O2F [M + H] + = 314.1305, found = 314.1309.

[0099] Compound 3ma, yellow solid, 58.9 mg, 89% yield, mp 249.6-250.9 °C. 1 H NMR (400 MHz, DMSO-d6) δ 13.27 (s, 1H), 8.09 - 7.57 (m, 2H), 7.22 (s, 1H), 7.01 (s, 1H), 5.17 (s, 1H), 3.65 (d, J = 22.3 Hz, 4H), 2.87 (s, 2H), 2.76 (s, 2H), 1.95 (s, 2H). 13 CNMR (101 MHz, DMSO-d6) δ 173.71, 172.71, 159.94 (d, J = 238.9 Hz), 152.12, 132.72 (d, J = 12.5 Hz), 131.93 (d, J = 3.6 Hz), 123.92 (d, J = 10.1 Hz), 122.17, 111.43 (d, J = 25.4 Hz), 98.85, 97.10 (d, J = 26.3 Hz), 96.05, 59.64, 46.47, 29.95, 27.14, 20.75. 17 H 17 N3O2Cl [M + H] + = 330.1009, found = 330.1013.

[0100] Compound 3na, yellow solid, 57.7 mg, 77% yield, mp 272.8-274.3 °C. 1H NMR (400 MHz, DMSO-d6) δ 13.28 (s, 1H), 8.00 (s, 1H), 7.72 (d, J = 8.6 Hz, 1H), 7.33 (d, J = 8.5 Hz, 1H), 7.00 (s, 1H), 5.17 (s, 1H), 3.68 (s, 2H), 3.66 - 3.58 (m, 2H), 2.88 (t, J = 5.8 Hz, 2H), 2.80 - 2.71 (m, 2H), 1.95 (p, J = 6.2 Hz, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 173.54, 173.00, 152.43, 133.48, 131.72, 125.03, 124.13, 115.99, 114.02, 99.17, 95.91, 59.61, 46.55, 29.94, 27.19, 20.73. HRMS (ESI) m / z calcd for C 17 H 17 N3O2Br [M+H] + = 374.0504, found = 374.0504.

[0101] Compound 3oa, yellow solid, 36.3 mg, 55% yield, mp 255.2-256.3 °C. 1 H NMR (400 MHz, DMSO-d6) δ 13.26 (s, 1H), 7.72 (d, J = 8.1 Hz, 1H), 7.33 (d, J = 7.4 Hz, 1H), 7.16 (t, J = 7.8 Hz, 1H), 7.09 (s, 1H), 5.17 (s, 1H), 3.75 - 3.57 (m, 4H), 2.88 (s, 2H), 2.75 (s, 2H), 1.96 (s, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 173.16, 172.89, 151.26, 132.30, 128.16, 128.11, 124.94, 122.29, 121.21, 118.03, 98.83, 96.77, 59.60, 46.55, 30.20, 27.19, 20.81. HRMS (ESI) m / z calcd for C 17 H 17 N3O2Cl [M+H] + = 330.1009, found = 330.1012.

[0102] Compound 3pa, yellow solid, 36 mg, 48% yield, mp 245.7-246.5 °C. 1H NMR (400 MHz, DMSO-d6) δ 13.24 (s, 1H), 7.77 (d, J = 8.0 Hz, 1H), 7.51 (d, J = 7.4 Hz, 1H), 7.15 - 7.03 (m, 2H), 5.17 (s, 1H), 3.73 - 3.57 (m, 4H), 2.87 (t, J = 5.7 Hz, 2H), 2.76 (t, J = 6.2 Hz, 2H), 1.96 (p, J = 6.3 Hz, 2H). 13 CNMR (101 MHz, DMSO-d6) δ 173.17, 172.87, 150.83, 132.31, 129.07, 128.43, 128.26, 122.77, 121.73, 104.70, 98.91, 96.63, 59.61, 46.54, 30.19, 27.19, 20.81. HRMS (ESI) m / z calcd for C 17 H 17 N3O2Br [M+H] + = 374.0504, found = 374.0506.

[0103] Compound 3qa, yellow solid, 38.2 mg, 62% yield, mp 190.7-191.9 °C. 1 H NMR (400 MHz, DMSO-d6) δ 13.27 (s, 1H), 7.55 (d, J = 7.2 Hz, 1H), 7.20 - 6.90 (m, 3H), 5.17 (s, 1H), 3.76 - 3.54 (m, 4H), 2.89 (s, 3H), 2.84 (s, 2H), 2.72 (s, 2H), 1.94 (s, 2H). 13 CNMR (101 MHz, DMSO-d6) δ 173.65, 172.44, 150.51, 131.76, 131.09, 125.89, 125.25, 124.26, 121.76, 119.68, 98.60, 96.36, 59.68, 46.43, 30.23, 27.13, 20.93, 20.08. HRMS (ESI) m / z calcd for C 18 H 20 N3O2 [M+H] + = 310.1556, found = 310.1559.

[0104] Compound 3ra, yellow solid, 57.4 mg, 85% yield, mp 275.3-276.6 °C. 1H NMR (400 MHz, DMSO-d6) δ 13.26 (s, 1H), 8.07 (d, J = 8.0 Hz, 1H), 7.73 (d, J = 7.4 Hz, 1H), 7.33 (t, J = 7.8 Hz, 1H), 7.23 (s, 1H), 5.19 (t, J = 4.7 Hz, 1H), 3.67 (dp, J = 15.7, 5.4 Hz, 4H), 2.90 (t, J = 6.2 Hz, 2H), 2.70 - 2.64 (m, 2H), 1.94 (p, J = 6.2 Hz, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 173.42, 173.05, 152.76, 137.82, 133.00, 128.10, 126.60, 121.43, 121.01, 118.39, 98.94, 97.10, 59.52, 46.66, 29.95, 27.24, 20.63. HRMS (ESI) m / z calcd for C 17 H 17 N4O4[M+H] + = 341.1250, found = 341.1246.

[0105] Compound 3sa, yellow solid, 66.2 mg, 90% yield, mp 265.4-266.6 °C. 1 H NMR (400 MHz, DMSO-d6) δ 13.27 (s, 1H), 7.82 (dd, J = 1.7, 0.9 Hz, 1H), 7.38 (d, J = 1.8 Hz, 1H), 6.92 (d, J = 0.9 Hz, 1H), 5.19 (t, J = 4.8 Hz, 1H), 3.67 (dp, J = 14.9, 4.6 Hz, 4H), 2.89 (t, J = 6.1 Hz, 2H), 2.80 - 2.72 (m, 2H), 1.96 (p, J = 6.0 Hz, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 173.37, 172.98, 153.40, 133.09, 132.57, 127.52, 127.02, 122.68, 121.68, 110.45, 99.40, 93.41, 59.54, 46.66, 29.88, 27.21, 20.62. HRMS (ESI) m / z calcd for C 17 H 16 N3O2Cl2[M+H] + = 364.0620, found = 364.0623.

[0106] Compound 3ta, yellow solid, 52.9 mg, 72% yield, mp 270.6-271.9 °C. 1 H NMR (400 MHz, DMSO-d6) δ 13.27 (s, 1H), 7.81 (s, 1H), 7.37 (s, 1H), 7.05 (s, 1H), 5.18 (s, 1H), 3.67 (d, J = 16.0 Hz, 4H), 2.90 (s, 2H), 2.75 (s, 2H), 1.96 (s, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 173.26, 172.75, 151.95, 133.48, 128.41, 126.74, 125.78, 124.21, 120.25, 119.15, 99.05, 96.37, 59.55, 46.65, 30.14, 27.24, 20.74. HRMS (ESI) m / z calcd for C 17 H 16 N3O2Cl2[M+H] + = 364.0620, found = 364.0620.

[0107] Compound 3ua, yellow solid, 66.0 mg, 89% yield, mp 243.2-244.3 °C. 1 H NMR (400 MHz, DMSO-d6) δ 13.18 (s, 1H), 7.82 (d, J = 1.9 Hz, 1H), 7.67 (d, J = 8.7 Hz, 1H), 7.29 (dd, J = 8.7, 1.9 Hz, 1H), 3.70 (t, J = 5.0 Hz, 2H), 3.63-3.58 (m, 2H), 2.85 (t, J = 6.2 Hz, 2H), 2.71 (t, J = 6.3 Hz, 2H), 1.93 (p, J = 6.4 Hz, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 173.05, 172.66, 152.83, 131.04, 129.25, 125.46, 123.36, 121.80, 120.40, 111.22, 99.42, 99.40, 59.53, 46.57, 29.79, 27.18, 20.52. HRMS (ESI) m / z calcd for C 17 H 16 N3O2Cl2[M+H] + = 364.0620, found = 364.0622.

[0108] Compound 3wa, white solid, 5.5 mg, 11% yield, mp 160.1-162.0 °C. 1 H NMR (400 MHz, DMSO-d6) δ 13.26 (s, 1H), 7.88 (d, J = 8.3 Hz, 1H), 7.75 (d, J = 8.0 Hz, 1H), 7.31 (t, J = 7.6 Hz, 1H), 7.21 (t, J = 7.5 Hz, 1H), 7.00 (s, 1H), 4.75 (s, 1H), 3.56 (q, J = 6.0 Hz, 4H), 2.78 (dt, J = 19.7, 6.2 Hz, 4H), 1.93 (p, J = 6.3 Hz, 2H), 1.81 (p, J = 6.6 Hz, 2H). 13 C NMR (126 MHz, CDC13) δ 171.65, 171.05, 152.31, 124.37, 119.28, 108.54, 104.44, 96.72, 59.93, 45.82, 28.15, 25.88, 19.42.

[0109] Compound 3ab, yellow solid, 54.8 mg, 88% yield, mp 173.3-174.3 °C. 1 H NMR (400 MHz, DMSO-d6) δ 13.26 (s, 1H), 7.88 (d, J = 8.3 Hz, 1H), 7.75 (d, J = 8.0 Hz, 1H), 7.31 (t, J = 7.6 Hz, 1H), 7.21 (t, J = 7.5 Hz, 1H), 7.00 (s, 1H), 4.75 (s, 1H), 3.56 (q, J = 6.0 Hz, 4H), 2.78 (dt, J = 19.7, 6.2 Hz, 4H), 1.93 (p, J = 6.3 Hz, 2H), 1.81 (p, J = 6.6 Hz, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 173.84, 172.58, 151.50, 133.13, 130.90, 125.35, 123.32, 122.17, 122.06, 111.62, 98.95, 95.69, 58.20, 40.96, 32.08, 29.92, 26.77, 20.79. HRMS (ESI) m / z calcd for C 18 H 20 N3O2[M+H] + = 310.1556, found = 310.1560.

[0110] Compound 3ac, yellow solid, 50.5 mg, 77% yield, mp 176.8-177.6 °C.1 H NMR (400 MHz, DMSO-d6) δ 13.27 (s, 1H), 7.88 (d, J = 8.4 Hz, 1H), 7.75 (d, J = 8.1 Hz, 1H), 7.31 (t, J = 7.6 Hz, 1H), 7.21 (t, J = 7.5 Hz, 1H), 6.99 (s, 1H), 4.55 (s, 1H), 3.50 (dt, J = 13.9, 6.6 Hz, 4H), 2.79 (dt, J = 18.0, 6.3 Hz, 4H), 1.94 (p, J = 6.1 Hz, 2H), 1.70 (p, J = 7.1 Hz, 2H), 1.56 (p, J = 6.9 Hz, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 173.88, 172.56, 151.49, 133.13, 130.86, 125.35, 123.34, 122.18, 122.08, 111.61, 98.93, 95.71, 60.65, 43.65, 30.08, 29.91, 26.82, 25.80, 20.79. HRMS (ESI) m / z calcd for C 19 H 22 N3O2[M+H] + = 324.1712, found = 324.1714.

[0111] Compound 3ad, yellow solid, 57.6 mg, 86% yield, mp 150.5-152.3 °C. 1 H NMR (400 MHz, DMSO-d6) δ 13.29 (s, 1H), 7.88 (s, 1H), 7.75 (s, 1H), 7.31 (s, 1H), 7.22 (s, 1H), 7.00 (s, 1H), 4.43 (s, 1H), 3.53 (s, 4H), 2.82 (d, J = 27.4 Hz, 4H), 1.96 (s, 2H), 1.68 (s, 2H), 1.47 (s, 4H). 13 C NMR (101 MHz, DMSO-d6) δ 173.90, 172.67, 151.51, 133.13, 130.85, 125.35, 123.36, 122.19, 122.10, 111.61, 98.95, 95.74, 60.97, 43.82, 32.48, 29.93, 28.88, 26.86, 23.39, 20.81. HRMS (ESI) m / z calcd for C 20 H 24 N3O2[M+H] += 338.1869, found = 338.1873.

[0112] Compound 3ae, yellow solid, 57.2 mg, 82% yield, mp 137.9-139.3 °C. 1 H NMR (400 MHz, DMSO-d6) δ 13.24 (s, 1H), 7.88 (d, J = 8.3 Hz, 1H), 7.75 (d, J = 8.0 Hz, 1H), 7.31 (t, J = 7.5 Hz, 1H), 7.21 (t, J = 7.4 Hz, 1H), 7.00 (s, 1H), 3.58 (s, 1H), 3.45-3.34 (m, 4H), 2.84-2.72 (m, 4H), 1.94 (p, J = 7.0, 6.1 Hz, 2H), 1.65 (p, J = 6.8 Hz, 2H), 1.52-1.36 (m, 6H). 13 C NMR (101 MHz, DMSO-d6) δ 173.88, 172.56, 151.48, 133.13, 130.85, 125.34, 123.34, 122.18, 122.07, 111.61, 98.93, 95.73, 61.08, 43.70, 32.85, 29.90, 28.98, 26.82, 26.64, 25.59, 20.78. HRMS (ESI) m / z calcd for C 21 H 26 N3O2[M+H] + = 352.2025, found = 352.2027.

[0113] Compound 3af, yellow solid, 58.8 mg, 81% yield, mp 133.4-135.8 °C. 1 H NMR (400 MHz, DMSO-d6) δ 13.25 (s, 1H), 7.87 (d, J = 8.3 Hz, 1H), 7.75 (d, J = 8.1 Hz, 1H), 7.31 (t, J = 7.7 Hz, 1H), 7.21 (t, J = 7.5 Hz, 1H), 7.00 (s, 1H), 4.36 (s, 1H), 3.55-3.39 (m, 4H), 2.79 (d, J = 16.2 Hz, 4H), 1.95 (s, 2H), 1.65 (s, 2H), 1.46-1.29 (m, 8H). 13C NMR (101 MHz, DMSO-d6) δ 173.89, 172.59, 151.48, 133.13, 130.85, 125.35, 123.34, 122.18, 122.08, 111.61, 98.93, 95.73, 61.14, 43.73, 32.94, 29.91, 28.98, 28.90, 26.84, 26.80, 25.89, 20.79. HRMS (ESI) m / z calcd for C 22 H 28 N3O2[M+H] + = 366.2182, found = 366.2184.

[0114] Compound 3ag, yellow solid, 39.2 mg, 64% yield, mp 169.4-170.0 °C. 1 H NMR (400 MHz, DMSO-d6) δ 13.29 (s, 1H), 7.87 (d, J = 8.3 Hz, 1H), 7.75 (d, J = 8.0 Hz, 1H), 7.34 - 7.28 (m, 1H), 7.24 - 7.18 (m, 1H), 6.99 (s, 1H), 3.54 (q, J = 6.2 Hz, 2H), 2.86 (t, J = 5.7 Hz, 2H), 2.80 - 2.74 (m, 2H), 1.96 (p, J = 6.1 Hz, 2H), 1.66 (p, J = 7.0 Hz, 2H), 1.44 (h, J = 7.4 Hz, 2H), 0.96 (t, J = 7.3 Hz, 3H). 13 C NMR (101 MHz, DMSO-d6) δ 173.90, 172.69, 151.50, 133.13, 130.85, 125.34, 123.35, 122.19, 122.09, 111.61, 98.96, 95.73, 43.45, 30.97, 29.92, 26.86, 20.81, 19.97, 14.01. HRMS (ESI) m / z calcd for C 19 H 22 N3O[M+H] + = 308.1763, found = 308.1767.

[0115] Compound 3ah, yellow oil, 6.6 mg, 11% yield, 1H NMR (400 MHz, DMSO-d6) δ 14.06 (s, 1H), 7.93 (d, J = 8.4 Hz, 1H), 7.81 (d, J = 8.0 Hz, 1H), 7.37 (t, J = 7.6 Hz, 1H), 7.27 (t, J = 7.4 Hz, 1H), 7.04 (s, 1H), 3.09 (t, J = 6.1 Hz, 2H), 2.86 (t, J = 6.0 Hz, 2H), 2.03 (t, J = 6.2 Hz, 2H), 1.59 (s, 9H). 13 C NMR (101 MHz, DMSO-d6) δ 173.80, 172.87, 151.38, 133.03, 130.63, 125.39, 123.32, 122.18, 122.12, 111.61, 99.31, 95.66, 55.06, 30.23, 30.16, 28.79, 21.41. HRMS (ESI) m / z calcd for C 19 H 22 N3O [M+H] + = 308.1763, found = 308.1767.

[0116] Compound 3ai, yellow solid, 30.8 mg, 53% yield, mp 137.1-138.9 °C. 1 H NMR (400 MHz, DMSO-d6) δ 13.14 (s, 1H), 7.90 (d, J = 8.3 Hz, 1H), 7.78 (d, J = 8.1 Hz, 1H), 7.35 (t, J = 7.6 Hz, 1H), 7.24 (t, J = 7.4 Hz, 1H), 7.03 (s, 1H), 3.10 (s, 1H), 3.04 (t, J = 5.6 Hz, 2H), 2.85-2.76 (m, 2H), 2.01 (p, J = 6.9 Hz, 2H), 1.00 (d, J = 7.2 Hz, 2H), 0.87 (d, J = 2.7 Hz, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 174.82, 174.06, 151.34, 133.25, 130.64, 125.33, 123.55, 122.24, 122.13, 111.62, 99.10, 96.17, 29.92, 27.72, 26.10, 20.83, 7.78. HRMS (ESI) m / z calcd for C 18 H 18 N3O [M+H] + = 292.1450, found = 292.1454.

[0117] Compound 3aj, yellow solid, 39.3 mg, 64% yield, mp 152.6-154.1 °C. 1 H NMR (400 MHz, DMSO-d6) δ 13.40 (d, J = 5.9 Hz, 1H), 7.87 (d, J = 8.3 Hz, 1H), 7.76 (d, J = 8.0 Hz, 1H), 7.32 (t, J = 7.6 Hz, 1H), 7.21 (t, J = 7.5 Hz, 1H), 7.01 (s, 1H), 4.38 (h, J = 7.8 Hz, 1H), 2.76 (t, J = 6.3 Hz, 4H), 2.47 - 2.38 (m, 2H), 2.18 - 2.07 (m, 2H), 1.93 (p, J = 6.3 Hz, 2H), 1.86 - 1.75 (m, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 174.05, 171.20, 151.47, 133.21, 130.72, 125.34, 123.48, 122.23, 122.12, 111.61, 98.80, 95.99, 48.18, 30.72, 29.88, 27.00, 20.80, 15.40. HRMS (ESI) m / z calcd for C 19 H 20 N3O[M+H] + = 306.1606, found = 306.1609.

[0118] Compound 3ak, yellow solid, 52.8 mg, 82% yield, mp 167.8-168.7 °C. 1 H NMR (400 MHz, DMSO-d6) δ 13.52 (d, J = 7.9 Hz, 1H), 7.88 (d, J = 8.3 Hz, 1H), 7.75 (d, J = 8.0 Hz, 1H), 7.36 - 7.27 (m, 1H), 7.26 - 7.17 (m, 1H), 6.99 (s, 1H), 4.26 (h, J = 7.4 Hz, 1H), 2.88 (t, J = 6.2 Hz, 2H), 2.82 - 2.73 (m, 2H), 2.11 - 2.00 (m, 2H), 2.02 - 1.89 (m, 2H), 1.82 - 1.73 (m, 2H), 1.71 - 1.59 (m, 4H). 13C NMR (101 MHz, DMSO-d6) δ 173.90, 171.77, 151.43, 133.14, 130.79, 125.36, 123.34, 122.19, 122.09, 111.62, 98.87, 95.71, 55.25, 33.58, 29.95, 27.13, 23.90, 20.89. HRMS (ESI) m / z calcd for C 20 H 22 N3O[M+H] + = 320.1763, found = 320.1768.

[0119] Compound 3al, yellow solid, 57.8 mg, 87% yield, mp 158.9-160.2 °C. 1 H NMR (400 MHz, DMSO-d6) δ 13.54 (d, J = 8.6 Hz, 1H), 7.88 (dd, J = 8.3, 1.0 Hz, 1H), 7.75 (d, J = 8.0 Hz, 1H), 7.31 (ddd, J = 8.2, 6.9, 1.1 Hz, 1H), 7.21 (ddd, J = 8.0, 6.9, 1.1 Hz, 1H), 7.00 (s, 1H), 3.93 - 3.80 (m, 1H), 2.91 (t, J = 6.2 Hz, 2H), 2.78 (dd, J = 7.1, 5.4 Hz, 2H), 1.96 (dt, J = 12.8, 7.1 Hz, 4H), 1.77 - 1.67 (m, 2H), 1.65 - 1.36 (m, 6H). 13 C NMR (101 MHz, DMSO-d6) δ 173.97, 171.34, 151.49, 133.13, 130.80, 125.37, 123.33, 122.19, 122.09, 111.63, 98.82, 95.70, 52.04, 32.63, 30.00, 26.60, 25.18, 23.91, 20.95. HRMS (ESI) m / z calcd for C 21 H 24 N3O[M+H] + = 334.1919, found = 334.1923.

[0120] The present application carries out fluorescence spectrum test on the synthesized target compound, and the organic reagents used in the test are chromatographically pure reagents (avoiding impurity peaks).

[0121] Application Example 1 tests the fluorescence performance of the target compound of the present application

[0122] The preparation of the test solution:

[0123] (1) Preparation of standard stock solution: Weigh a certain amount of 7-amino-4-trifluoromethylcoumarin and dissolve it in methanol (chromatographic grade). Shake evenly to prepare a concentration of 10 -3 The test stock solution of M is further diluted when the specific test is performed.

[0124] (2) Preparation of test sample mother solution: Dissolve the synthesized target compound in an organic solvent to prepare 10 -3 The stock solution of M is further diluted when testing.

[0125] (3) The test solvent was methanol (chromatographic grade).

[0126] (4) Fluorescence data testing requires the preparation of solutions at 10 μM, 5 μM, 2.5 μM, 0.625 μM, 0.3125 μM, and 0.156 μM. This involves diluting 100 μL of the stock solution to 10 mL with the desired organic reagent. The solution is then diluted in half to create a gradient of sample solutions. The test is performed at room temperature.

[0127] According to the Beer-Lambert law, fluorescence spectroscopy was used to detect the fluorescence intensity of substances at different concentrations. -5 The fluorescence spectrum of M in methanol solution is as follows Figure 3 As shown, the scanning range of the fluorescence spectrometer is 420-800nm, and the scanning speed is 2000nm / min. The normalized fluorescence emission spectra of some compounds are shown in Figure 4 As shown in the figure, it was found that the maximum fluorescence emission wavelength of indole-fused hydrogenated cinnolines is related to the electronic effect of the substituents, and the fluorescence wavelength range of this type of compound is between 430-700nm. Compared with the template product 3aa without substituents, it can be found that most products with electron-withdrawing groups on the benzene ring (3ba, 3ca, 3da, 3fa, 3ga, 3ha, 3ka, 3la, 3ma, 3na, 3oa, 3pa, 3sa, 3ta, 3ua) will undergo a slight blue shift. On the other hand, compounds with electron-donating groups (3ea, 3ia, 3ja) will undergo a red shift. In addition, compared with compounds containing electron-withdrawing groups, they will exhibit weaker fluorescence ( Figure 3 The nitro-substituted compound 3ra was also found to lack fluorescence, likely due to the strong electron-withdrawing ability of the nitro group, which disrupts the compound's conjugated structure, leading to fluorescence quenching. Extending the hydroxyethyl carbon chain resulted in a slight red-shift in the compounds (3ab, 3ac, 3ad, 3ae, and 3af). Similarly, replacing the chain structure with a cyclic structure resulted in a slight red-shift in the compounds (3ai, 3aj, 3ak, and 3al).

[0128] The fluorescence quantum yield of the target compounds in methanol solution was calculated with 7-amino-4-trifluoromethyl coumarin as the reference substance and its quantum yield (Ф = 0.53) as the standard. According to the literature reference, the λ em = 480 nm, λ ex = 376 nm, the test results of the present application are λ em = 484.3 nm, λ ex = 381.1 nm Figure 5 The fluorescence excitation spectrum and emission spectrum were made, the concentration curve was made, and the fluorescence quantum yield of each target compound was obtained. The results are shown in Table 4. It can be seen that the quantum yield of the target compounds is in the range of 15.3% to 72.9%, and the product with the highest quantum yield is 3pa, which can reach 72.9%. The compound 3ja has a large Stokes shift, reaching 100 nm. These experimental results show that simple modification of the structure of indole-fused hydrogenophan compounds can change their fluorescence properties. The above results prove that the indole-fused hydrogenophan compounds of the present application are expected to play their value in the fields of fluorescent materials, fluorescent imaging probes, etc.

[0129] Table 4 Optical properties of compounds in methanol solution

[0130]

[0131] Application Example 2 Test of Anti-tumor performance of the compounds of the present application

[0132] 1. Preparation of common reagents:

[0133] (1) Complete culture solution: 44.5 mL of cell culture was taken into a 50 mL centrifuge tube, 5 mL of fetal bovine serum and 0.5 mL of ampicillin-streptomycin mixture were added, and it was mixed well and stored in a 4°C box for use.

[0134] (2) Cell cryopreservation solution: The volume ratio of cell solution:fetal bovine serum:DMSO was 7:2:1, and the general cryopreservation cell solution volume of each cryopreservation tube was 1 mL. The cryopreservation solution was prepared and used immediately.

[0135] (3) MTT solution: 25 mg of MTT solid was taken into a 50 mL centrifuge tube, diluted to 50 mL with PBS buffer solution, mixed well, fully dissolved, and the concentration was 5 mg / mL. It was sterilized by passing through a 0.22 μM microporous filter and was divided into 2 mL light-proof centrifuge tubes and stored at -20°C in the dark. The whole preparation process was carried out in the dark as much as possible.

[0136] (4) Drug gradient concentration; for example, 50 μM, 25 μM, 12.5 μM, 6.25 μM, 3.125 μM, 1.56 μM, 0.78 μM, 0.39 μM, containing 5% DMSO (dimethyl sulfoxide).

[0137] 2. Experimental method

[0138] 2.1 Cell culture

[0139] (1) Cell recovery: take the frozen cells from liquid nitrogen or -80°C refrigerator, quickly place in 37°C water bath and gently shake to thaw, after alcohol disinfection of the frozen tube, place in the biological safety cabinet, transfer the cell liquid to the cell culture bottle containing appropriate amount of complete medium with a disposable pipette, mix well by blowing, and place in a 37°C, 5% CO2 incubator until it adheres, and replace the liquid after 24 hours of culture.

[0140] (2) Cell replacement: after observing the cell adhesion under a microscope, transfer the cell culture bottle to the biological safety cabinet, discard the old medium and rinse the cells with PBS 3 times, and add appropriate amount of new complete medium.

[0141] (3) Cell passage: take out the cells in the incubator and observe under a microscope, when the cells grow to 80%-90% of the bottom of the culture bottle, the cells can be passaged; aspirate the old culture medium, rinse the cells with PBS 3 times, add appropriate amount of trypsin-EDTA digestion solution to completely cover the cell layer of the culture bottle, at this time the cells can be incubated in a 37°C cell culture box for 1-2 min, and the cell shedding is observed under a microscope; when the cell shedding degree reaches 80%-90%, add appropriate amount of complete medium to terminate digestion, rinse the cell bottle bottom with a disposable pipette to make the cells completely shed, transfer the cells to a 15 mL centrifuge tube, centrifuge at 1000 r / min for 5 min, discard the supernatant, add appropriate amount of culture medium, mix well by blowing, and then divide into bottles for culture, add appropriate amount of complete medium, and place in a 37°C, 5% CO2 incubator.

[0142] (4) Cell freezing: centrifuge the cells in the logarithmic growth phase after digestion, discard the supernatant, add appropriate amount of cell freezing solution, mix the cells by blowing, divide into cell freezing tubes, place in a programmed cooling box, transfer to a -80°C refrigerator overnight, and finally transfer to liquid nitrogen for storage.

[0143] 2.2 MTT method for determining compound IC 50

[0144] MTT, also known as thiazole blue, is a yellow powder commonly used for cell proliferation and activity detection. The principle of detection is that succinate dehydrogenase in mitochondria can reduce MTT into insoluble blue-purple crystals, which are called formazan. The enzymes in dead cells are inactivated and cannot form the crystals. The formed crystals can be dissolved in dimethyl sulfoxide, and the absorbance is detected by an enzyme-labeled instrument at a specific wavelength. According to the measured absorbance, the survival rate of cancer cells can be reflected. The obtained absorbance is processed and calculated using analysis software, and the cell proliferation inhibition of the compound can be directly obtained, which is conducive to the analysis and summary of the efficacy of the compound.

[0145] (1) A plate: take the liver cancer HepG2, lung cancer A549, breast cancer MDA-MB-231, prostate cancer Du145, and colon cancer HCT-116 cells in the logarithmic growth phase, respectively, and use an appropriate amount of trypsin to digest. When the degree of cell shedding is 80%-90%, stop digestion. Collect the cells, centrifuge at 1500 r / min for 5 min, resuspend the cells with an appropriate amount of complete medium, and count by a cell counter after uniform blowing. Dilute the cell solution to 5x10 4 6 / ml with complete medium, then inoculate the cell suspension into a 96-well plate at 100 uL per well, and culture in a 37℃, 5% CO2 incubator for 24 h.

[0146] (2) Drug administration: discard the supernatant in the 96-well plate, and add 100 uL of medium containing compound at concentrations of 50 uM, 25 uM, 12.5 uM, 6.25 uM, 3.125 uM, 1.56 uM, 0.78 uM, and 0.39 uM to the drug administration group, respectively. The blank group is added with the same volume of the same medium containing 5‰ DMSO, and the negative control group is added with the same volume of the same medium containing 5‰ DMSO. Each group has six replicate wells, and is cultured in a 37℃, 5% CO2 incubator for 48 h.

[0147] (3) Detection: add 10 uL of 5 mg / mL MTT solution to each well, continue to culture at 37℃ for 4 h, discard the supernatant, add 100 uL of DMSO to each well, mix on a plate shaker, and then detect the OD value by an enzyme-labeled instrument with a detection wavelength of 490 nm.

[0148] (4) Tumor cell growth inhibition rate calculation formula: cell inhibition rate (%) = [1-(OD (药物组) -OD (调零组) / (OD (空白组) -OD (调零组) ]x100%.

[0149] 2.3 Data processing

[0150] The original data were processed by Excel to obtain the growth inhibition rates at different concentrations, and the IC values of each compound were calculated by GraphPad Prism6. 50 values.

[0151] 3.3 Experimental results

[0152] 3.3.1 Purity detection of indole-fused hydrocarbazole compounds

[0153] (1) High performance liquid chromatography (HPLC) conditions

[0154] Column: Gemini 5 μm NX-C18; mobile phase: methanol-water (90 mL / 10 mL); detection wavelength: 254 nm, 365 nm; flow rate: 1.0 mL / min; column temperature: 30°C; injection volume: 10 μL.

[0155] (2) High performance liquid chromatography (HPLC) determination method

[0156] The synthesized target compound indole-fused hydrocarbazole compound was dissolved in methanol, and the dissolved sample was filtered through a 0.22 μm filter to remove insoluble substances. The filtered solution was transferred to a liquid phase vial, and the purity was determined by HPLC.

[0157] The purity of the indole-fused hydrocarbazole compounds of the present application is shown in Table 5.

[0158] Table 5 Purity of indole-fused hydrocarbazole compounds

[0159]

[0160]

[0161] A series of indole-fused hydrocarbazole derivatives were obtained, and the structure of the compounds was confirmed to be consistent with the structure of the expected target compounds after identification by hydrogen spectrum, carbon spectrum and high resolution mass spectrometry. The synthesized compounds were all subjected to HPLC purity determination, and the purity of all the compounds was more than 95%.

[0162] 3.3.2 In vitro anti-tumor activity of target compounds

[0163] Among the synthesized indole-fused hydrocarbazole compounds, only a few compounds showed good in vitro proliferation inhibition activity against lung cancer A549, breast cancer MDA-MB-231, colon cancer HCT116, liver cancer HepG2 and prostate cancer DU145 cell lines. The specific results are shown in Table 5.

[0164] Table 5 Summary of in vitro anti-tumor activity of target compounds

[0165]

[0166]

[0167] The present application investigates the effect of different substituents on the indole benzene ring on the cell proliferation inhibition of five cancer cell lines. As can be seen from Table 5, when an electron-withdrawing group -Br is introduced at position 4, compound 3da shows good anti-tumor activity against five cancer cells A549, MDA-MB-231, HCT116, HepG2 and DU145, and the IC 50 values are 20.48 μM, 17.63 μM, 7.498 μM, 11.29 μM and 18.98 μM, respectively. When different substituents are introduced at position 4 or other positions, the IC 50 values of most of the monosubstituted products obtained are greater than 50 μM, so it can be seen that the introduction of different substituents has a great effect on the inhibition of cancer cells. It can also be concluded that when the indole ring is substituted with halogen, the compound shows certain biological activity. When an electron-withdrawing group -Br is also introduced at position 7 of the indole benzene ring, 3pa inhibits the proliferation of the remaining four cells in addition to A549.

[0168] On the basis of the study of monosubstitution of the indole benzene ring, two chlorine atoms are introduced into the indole benzene ring to obtain compounds containing dichloro substitution on the indole benzene ring (3sa, 3ta, 3ua). On this basis, the anti-tumor effect of the dihalogen-substituted compound is compared with that of the bromine monosubstituted compound 3da. In general, 3sa and 3ua containing dichloro substitution have better cell proliferation inhibition effect on cancer cell lines than 3da and 3pa, but the IC 50 value of compound 3ta with chlorine atoms introduced at positions 5 and 7 is greater than 50 μM. This shows that the cell viability of the five cancer cells is affected by the different positions of the substituents of this type of compound, and the inhibitory effect on cancer cells is better when substituents are introduced at positions 4 and 7 than when substituents are introduced at other positions. It is speculated that the introduction of substituents at positions 4 and 7 increases the steric hindrance of the compound, thereby affecting the inhibitory effect of the compound on cancer cells.

[0169] Only four compounds, 3da, 3pa, 3sa and 3ua, in this system have good proliferation inhibition effect on the five cancer cells, and the remaining compounds have no obvious cell proliferation inhibition effect on these cancer cells, indicating that these four compounds have certain universality for cancer cells. However, it is also found that there are more than four compounds that have inhibitory effect on colon cancer HCT116, suggesting that the inhibitory effect of the compounds on tumor cells has certain cell selectivity.

[0170] In the study of the effect of introducing substituents at different positions of the indole benzene ring on the activity of colon cancer HCT116, it was found that when electron-withdrawing groups -Cl or -Br were introduced at positions 4, 5 and 7, the compounds (3ca, 3da, 3ga, 3ha, 3oa, 3pa) all showed good cancer cell inhibition effect, and the obtained IC 50 values were all less than 50 μM. However, the compounds (3ma, 3na) with -Cl or -Br introduced at position 6 had less inhibitory effect on colon cancer HCT116 than the compounds with substituents introduced at other positions. This indicates that introducing halogen atoms at position 6 is not conducive to obtaining compounds with superior antitumor activity. Then, the inhibitory effect of compounds with electron-donating groups on colon cancer HCT116 was investigated. When electron-donating groups -Me were introduced at different positions of the indole benzene ring, the compounds (3ea, 3ia) did not show cytotoxicity to colon cancer HCT116. However, 3ja showed certain inhibitory effect when -OMe was introduced at position 5, and the IC 50 value was 19.68 μM, indicating that substituents of different properties have a great influence on cytotoxicity.

[0171] The present application investigated the inhibitory effect of compounds obtained by modifying the structure of hydrocinnoline on colon cancer HCT116. After extending the carbon chain at position 5 of hydrocinnoline, 3ab-3af still did not show obvious cytotoxicity. However, after replacing the chain hydrocarbon with a cycloalkane, 3ak showed certain cell proliferation inhibition effect, and the IC 50 value was 33.77 μM, which is presumably related to steric hindrance.

[0172] The present application mainly uses the MTT method to study the in vitro antitumor activity of the obtained target indole-fused hydrocinnoline compounds. The absorbance (OD value) of the cells is determined, and according to the linear relationship between the absorbance and the survival of the cells, the OD values measured are processed, analyzed and the IC 50 values are calculated by analysis software, and the calculated IC 50 values are obtained. The structure-activity relationship of the compounds is summarized. Overall, the indole-fused hydrocinnoline compounds of the present application have better cell inhibition activity on colon cancer HCT116.

[0173] The above is only a preferred specific embodiment of the present application, but the protection scope of the present application is not limited thereto. Any changes or replacements easily thought of by those skilled in the art within the technical scope disclosed by the present application should be covered within the protection scope of the present application. Therefore, the protection scope of the present application should be subject to the protection scope of the claims.

Claims

1. An indole-fused hydrogenated cinnoline compound, characterized in that: The structural formula is as follows: wherein R is selected from H, halogen, methyl, methoxy, cyano or nitro, R 1 Selected from hydroxyalkyl or alkyl.

2. The indole-fused hydrocinnoline compound according to claim 1, characterized in that When the R is a halogen, it includes monohalogen substitution and dihalogen substitution; the hydroxyalkyl group is a C2-C7 hydroxyalkyl group; the alkyl group is n-butyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

3. The indole-fused hydrocinnoline compound according to claim 2, characterized in that Selected from the following compounds:

4. A method for preparing the indole-fused hydrocinnoline compound according to any one of claims 1 to 3, characterized in that: The indole-fused hydrogenated cinnoline compound is synthesized by using an enaminone compound and an amine compound as reaction substrates in a highly polar solvent through a stork enamine reaction and an intramolecular acylation reaction; The structural formula of the enaminone compound is wherein R is selected from H, halogen, methyl, methoxy, cyano or nitro; The structural formula of the amine compound is R 1 Selected from hydroxyalkyl or alkyl.

5. The method for preparing the indole-fused hydrocinnoline compound according to claim 4, wherein: The molar ratio of the enaminone compound to the amine compound is 1:(1-10).

6. The method for preparing the indole-fused hydrocinnoline compound according to claim 4, wherein: The temperature of the stork enamine reaction and intramolecular acylation reaction is 45-80°C.

7. The method for preparing the indole-fused hydrocinnoline compound according to claim 4, wherein: The highly polar solvent is selected from one of methanol, acetonitrile and chloroform.

8. Use of the indole-fused hydrogenated cinnoline compound according to any one of claims 1 to 3 in the preparation of fluorescent materials and / or semiconductor materials.

9. Use of the indole-fused hydrocinnoline compound according to any one of claims 1 to 3 in the preparation of anti-tumor drugs.

10. The use according to claim 9, characterized in that The anti-tumor drugs include drugs for preventing and / or treating lung cancer, drugs for preventing and / or treating breast cancer, drugs for preventing and / or treating liver cancer, and drugs for preventing and / or treating prostate cancer.

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