Carbazole / benzo [cd] indole compound as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of biological medicines, and discloses a carbazole / benzo [cd] indole compound as well as a preparation method and application thereof. The carbazole / benzo [cd] indole compound has a structure as shown in a general formula I: # imgabs0 #, wherein R is one of NO2, NH2, Br and N (Ph) 2. The carbazole / benzo [cd] indole compound can generate active oxygen molecules after illumination, including superoxide anions and hydroxyl radicals, and can be applied to preparation of drugs for photodynamic therapy.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and particularly relates to a carbazole / benzo[cd]indole compound, a preparation method thereof, and an application thereof. Background Art

[0002] Cancer is a common cause of high morbidity and mortality worldwide. Although surgery remains the main method for treating cancer, chemotherapy with anticancer drugs is necessary in cases of residual cancer cells after surgery and advanced tumors to reduce the risk of tumor recurrence. However, traditional chemotherapy has some drawbacks, including severe side effects, insufficient efficacy, and challenges related to drug resistance. Therefore, single therapies may not be effective in treating tumors, and combined and / or complementary treatment modalities need to be explored.

[0003] Photodynamic therapy (PDT) has advantages due to its spatiotemporal precision, controllability, low drug resistance, and minimal side effects, making it superior to traditional treatment methods. Type-I photosensitizers (PSs) that do not rely on oxygen can effectively utilize disproportionation reactions, Haber-Weiss reactions, or Fenton reactions to relieve oxygen depletion by generating superoxide radicals (·O2 - ) or hydroxyl radicals (·OH). This strategy addresses the limitations of PDT in hypoxic tumors, thereby significantly improving the treatment effect. At the same time, further integration of chemotherapy and / or immunotherapy will enhance the treatment effect of PSs on hypoxic-resistant malignant tumors.

[0004] Carbazole and its derivatives, as natural products, exhibit various biological activities, such as anticancer, anti-inflammatory, and antiviral effects. The electron-donating properties of the carbazole core have been effectively used in the design of PSs. However, the limited antitumor effects of existing compounds have led to limited research on independent type-I carbazole-based PSs. Therefore, a comprehensive examination of the carbazole nucleus is crucial for evaluating its potential in type-I photodynamic therapy and chemotherapy. Summary of the Invention

[0005] The purpose of the present invention is to solve the defects existing in the prior art, and to provide a class of carbazole / benzo[cd]indole compounds, which have effective photodynamic therapy activity.

[0006] To achieve the above purpose, the present invention provides the following technical solutions:

[0007] In the first aspect of the present invention, there is provided a carbazole / benzo[cd]indole compound, and the carbazole / benzo[cd]indole compound has a structure shown in general formula I:

[0008]

[0009] Wherein, R is one of NO2, NH2, a halogen atom and (Ph)2N. The halogen atom is preferably Br.

[0010] The code numbers of the compounds of the above general formula I and their corresponding compound names are as follows:

[0011] When R is NO2, I1: (E)-2-(2-(9-ethyl-6-nitro-9H-carbazol-3-yl)vinyl)-1-methylbenz[cd]indole-1-iodine salt.

[0012] When R is NH2, I2: (E)-2-(2-(6-amino-9-ethyl-9H-carbazol-3-yl)vinyl)-1-methylbenz[cd]indole-1-iodine salt.

[0013] When R is Br, I3: (E)-2-(2-(6-bromo-9-ethyl-9H-carbazol-3-yl)vinyl)-1-methylbenz[cd]indole-1-iodine salt.

[0014] When R is (Ph)2N, I4: (E)-2-(2-(6-(diphenylamino)-9-ethyl-9H-carbazol-3-yl)vinyl)-1-methylbenz[cd]indole-1-iodine salt.

[0015] In a second aspect of the present invention, the present invention further provides a method for preparing the above-mentioned carbazole / benzo[cd]indole compound, the preparation method being as follows:

[0016] N-ethylcarbazole-3-carboxaldehyde (1) is subjected to nitration reaction to obtain 9-ethyl-6-nitro-9H-carbazole-3-carboxaldehyde (2). Compound (2) and 1,2-dimethylbenz[cd]indol-1-ium undergo Knoevenagel condensation reaction in the presence of a catalytic amount of piperidine to obtain compound I1.

[0017] Compound (1) is subjected to NBS bromination reaction to obtain compound (3). Compound (3) and 1,2-dimethyl-1H-benzo[cd]indol-1-ium undergo Knoevenagel condensation reaction in the presence of a catalytic amount of piperidine to obtain compound I3.

[0018] Compound (1) is subjected to NBS bromination to obtain compound (3). Compound (3) and diphenylamine are reacted in the presence of palladium acetate, tri-tert-butylphosphine, and cesium carbonate to obtain compound (4). Compound (4) undergoes a Knoevenagel condensation reaction with 1,2-dimethyl-1H-benzo[cd]indol-1-ium in the presence of a catalytic amount of piperidine to obtain compound I4.

[0019] Compound I1 is heated to reflux in the presence of iron powder and ammonium chloride in methanol solvent, and undergoes a reduction reaction to obtain compound I2.

[0020] Compounds I1, I2, I3 and I4 all belong to General Formula I.

[0021] The synthetic route diagram is as follows:

[0022]

[0023] In a third aspect of the present invention, there is provided the use of the above-mentioned carbazole / benzo[cd]indole compound in the preparation of a drug or a reagent for photodynamic therapy of a disease;

[0024]

[0025] wherein, R is selected from one of NO2, NH2, Br and N(Ph)2.

[0026] In some embodiments of the present invention, R is selected from NO2, NH2, Br and N(Ph)2, and the disease is a tumor.

[0027] In some embodiments of the present invention, the tumor is selected from: colon cancer, lung cancer or breast cancer.

[0028] In some embodiments of the present invention, the disease is a disease related to bacterial infection.

[0029] In some embodiments of the present invention, the bacteria are selected from: Staphylococcus aureus or methicillin-resistant Staphylococcus aureus.

[0030] In some embodiments of the present invention, the photodynamic therapy is carried out by a method comprising the following steps:

[0031] S1. Administering the carbazole / benzo[cd]indole compound to a subject; and

[0032] S2. Irradiating the subject with light having a wavelength of 450 - 520 nm.

[0033] In some embodiments of the present invention, the method has one or more of the following features:

[0034] (1) In step S2, the irradiation time is 3 - 10 min;

[0035] (2) In step S2, irradiating with a light intensity of 50 - 200 mW / cm 2 of light.

[0036] The present invention has the following advantages compared with the prior art:

[0037] The compounds of the present invention can all generate significant amounts of reactive oxygen species (ROS). Among them, compound I1 mainly generates type I ROS, such as superoxide anion (·O2-) and hydroxyl radical (●OH), which are very effective in overcoming the oxygen dependence in PDT. Compound I1 exhibits significant anti-proliferative activity in both normoxic and hypoxic environments, highlighting its oxygen-independent chemo / photodynamic therapy potential. In vivo experiments show that the compounds of the present invention can effectively inhibit tumor growth, with an inhibition rate reaching 93.1%. Description of the Drawings

[0038] Figure 1 It is the fluorescence emission spectrum diagram of the production of reactive oxygen species by the carbazole / benzo[cd]indole compounds I1, I2, I3 and I4 of the present invention. The abscissa is the wavelength and the ordinate is the fluorescence intensity;

[0039] Figure 2 It is the fluorescence emission spectrum diagram of the production of superoxide anion by the carbazole / benzo[cd]indole compounds I1, I2, I3 and I4 of the present invention. The abscissa is the wavelength and the ordinate is the fluorescence intensity;

[0040] Figure 3 It is the ultraviolet absorption spectrum diagram of the production of singlet oxygen by the carbazole / benzo[cd]indole compounds I1, I2, I3 and I4 of the present invention. The abscissa is the wavelength and the ordinate is the absorbance value;

[0041] Figure 4 It is the fluorescence emission spectrum diagram of the production of hydroxyl radical by the carbazole / benzo[cd]indole compounds I1, I2, I3 and I4 of the present invention. The abscissa is the wavelength and the ordinate is the fluorescence intensity;

[0042] Figure 5 It is the in vivo anti-tumor activity test diagram of the carbazole / benzo[cd]indole compound I1 of the present invention. Detailed Embodiments

[0043] The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings, so that those skilled in the art can better understand the advantages and features of the present invention, and thus make a clearer definition of the protection scope of the present invention. The embodiments described in the present invention are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0044] Example 1: Preparation of (E)-2-(2-(9-ethyl-6-nitro-9H-carbazol-3-yl)vinyl)-1-methylbenzo[cd]indole-1-iodide (I1)

[0045] 9-Ethyl-6-nitro-9H-carbazole-3-carbaldehyde (2) (1 g, 3.73 mmol) and 1,2-dimethylbenzo[cd]indol-1-ium (1.15 g, 3.73 mmol) were dissolved in anhydrous ethanol (25 mL). After adding two drops of pyridine as a base, the mixture was heated at 80 °C for 30 minutes. After the reaction was completed, the solvent was evaporated using a rotary evaporator, and the crude product was purified by column chromatography using dichloromethane as an eluent to obtain the purified blue-green solid I1 with a yield of 65%.

[0046] The spectral data of compound I1 are as follows: 1 1H NMR (400 MHz, DMSO-d6) δ 9.37 (d, J = 7.3 Hz, 1H, ArH), 9.03 (s, 1H, ArH), 8.67 (d, J = 7.9 Hz, 1H, ArH), 8.41 - 8.34 (m, 2H, ArH), 8.17 (t, J = 7.7 Hz, 1H, ArH), 8.08 - 8.04 (m, 2H, ArH), 8.01 (s, 1H, ArH), 7.94 (t, J = 7.8 Hz, 1H, ArH), 7.87 (d, J = 8.7 Hz, 1H, ArH), 7.75 (d, J = 8.1 Hz, 1H, ArH), 7.61 - 7.57 (m, 1H, CH=CH), 7.41 - 7.37 (m, 1H, CH=CH), 4.55 (q, J = 7.2 Hz, 2H, CH2), 4.03 (s, 3H, CH3), 1.39 (t, J = 7.0 Hz, 3H, CH3).

[0047] 13 13C NMR (101 MHz, CDCl3) δ 156.0, 149.0, 144.7, 143.8, 141.6, 136.1, 131.9, 130.0, 128.4, 127.1, 124.9, 124.6, 123.1, 122.8, 122.5, 120.6, 117.6, 117.1, 115.8, 109.9, 109.0, 101.5, 38.7, 23.5, 13.9.

[0048] Example 2: Preparation of (E)-2-(2-(6-amino-9-ethyl-9H-carbazol-3-yl)vinyl)-1-methylbenzo[cd]indol-1-ium iodide (I2)

[0049] Compound I1 (500 mg, 0.894 mmol) was dissolved in ethanol (3 mL), and then saturated ammonium chloride solution (1 mL) and iron powder (10 mmol) were added. The reaction was carried out for 0.5 h. After completion, the residual iron powder was removed by washing with methanol. The mixture was concentrated under reduced pressure, and the crude product was purified by column chromatography using dichloromethane and methanol (5:1, v / v) as the eluent to obtain green solid I2 with a yield of 55%.

[0050] The spectral data of compound I2 are as follows: 1 1H NMR (400 MHz, DMSO-d6) δ 9.37 (d, J = 7.4 Hz, 1H, ArH), 9.13 - 9.05 (m, 1H, ArH), 8.67 (d, J = 7.9 Hz, 1H, ArH), 8.53 - 8.33 (m, 1H, ArH), 8.29 - 8.24 (m, 2H, 2ArH), 8.17 (t, J = 7.7 Hz, 1H, ArH), 8.05 - 8.01 (m, 1H, ArH), 7.94 (t, J = 7.8 Hz, 1H, ArH), 7.87 (d, J = 8.7 Hz, 1H, ArH), 7.75 (d, J = 8.1 Hz, 1H, ArH), 7.61 - 7.57 (m, 1H, CH=CH), 7.41 - 7.37 (m, 1H, CH=CH), 4.55 (q, J = 7.2 Hz, 2H, CH2), 4.32 (s, 2H, NH2), 4.03 (s, 3H, CH3), 1.39 (t, J = 7.0 Hz, 3H, CH3).

[0051] 13 13C NMR (101 MHz, CDCl3) δ 191.7, 143.7, 139.3, 129.4, 128.8, 127.6, 124.7, 124.5, 123.6, 122.0, 113.2, 110.6, 109.1, 38.1, 13.8

[0052] Example 3: Preparation of (E)-2-(2-(6-bromo-9-ethyl-9H-carbazol-3-yl)vinyl)-1-methylbenzo[cd]indol-1-ium iodide (I3)

[0053] In a Schlenk flask, compound 4 (1 g, 3.31 mmol) and 1,2-dimethyl-1H-benzo[cd]indol-1-ium (1.02 g, 3.31 mmol) were dissolved in anhydrous ethanol (25 mL). After adding two drops of pyridine as a base, the mixture was heated at 80 °C for 30 minutes. After the reaction was completed, the solvent was evaporated under reduced pressure. The crude product was purified by column chromatography using dichloromethane and methanol (5:1, v / v) as the eluent to obtain blue-green solid I3 with a yield of 68%.

[0054] The spectral data of Compound I3 are as follows: 1 H NMR(400MHz,DMSO-d6)δ8.89(t,J=1.4Hz,1H,ArH),8.73-8.69(m,1H,ArH),8.63(s,1H,ArH),8.50-8.46(m,1H,ArH),8.31(d,J=8.9Hz,1H,ArH),8.27-8.22(m,2H,ArH),8.16(d,J=8.9Hz,1H,ArH),8.00-7.97(m,1H,ArH),7.83-7.81(m,1H,ArH),7.76-7.71(m,2H,ArH),7.66-7.64(m,1H,CH=CH),7.43-7.40(m,1H,CH=CH),4.28-4.22(m,2H,CH2),2.93(s,3H,CH3),2.12-2.16(m,3H,CH3)

[0055] 13 C NMR(101MHz,CDCl3)δ191.7,182.7,175.2,143.7,139.3,129.4,128.8,127.6,124.7,124.4,123.6,122.0,113.2,111.1,110.7,110.4,109.1,109.0,104.8,99.8,38.2,24.4,14.3,13.8

[0056] Example 4: (E)-2-(2-(6-(Diphenylamino)-9-ethyl-9H-carbazol-3-yl)vinyl)-1-methylbenzo[cd]indol-1-ium

[0057] Compound 4(500mg,1.28mmol) and 1,2-dimethylbenzo[cd]indol-1-ium(395mg,1.28mmol) were dissolved in 25mL of a Schlenk tube with absolute ethanol, 2 drops of piperidine were used as the base, and the reaction was carried out at 80°C for 30min. After the reaction was completed, the solvent was evaporated, and column chromatography was used with dichloromethane:methanol(5:1, v / v) as the eluent to purify to obtain a blue-green solid I4 with a yield of 65%.

[0058] The spectral data of Compound I4 are as follows: 11H NMR (400 MHz, DMSO-d6) δ 8.78 (m, 1H, ArH), 8.72 (d, J = 1.5 Hz, 1H, ArH), 8.08 (d, J = 2.0 Hz, 1H, ArH), 8.00 - 7.95 (m, 2H, 2ArH), 7.78 (t, J = 7.7 Hz, 2H, 2ArH), 7.72 (m, 2H, 2ArH), 7.38 (d, J = 1.1 Hz, 1H, ArH), 7.32 (d, J = 2.2 Hz, 2H, 2ArH), 7.31 - 7.24 (m, 5H, 5ArH), 7.12 (m, 1H, CH=CH), 7.01 (m, 5H, 5ArH), 6.97 (d, J = 7.2 Hz, 1H, CH=CH), 4.50 (q, J = 6.7 Hz, 2H, CH2), 2.58 (s, 3H, CH3), 1.37 (t, J = 7.1 Hz, 3H, CH3).

[0059] Example 5: Test on the ability of the carbazole / benzo[cd]indole compound of the present invention to generate reactive oxygen species

[0060] The fluorescence spectrometry was used to detect the ability of the compound of the present invention to generate reactive oxygen species. 2,7-Dichlorodihydrofluorescein (DCFH) was used as a scavenger for reactive oxygen species. The specific method was as follows: A solution of the compound of the present invention (10 μM) and the scavenger DCFH (10 μM) was mixed, and then irradiated with a laser at 520 nm (50 mW / cm 2 ) for a total of 3 min, and the test was carried out at intervals of 20 s. The fluorescence spectrometer was used to detect the change in the fluorescence emission spectrum of the compound of the present invention at 523 nm at different times after laser irradiation. Figure 1 The fluorescence emission spectra of the compounds I1, I2, I3 and I4 of the present invention for generating reactive oxygen species are shown. The abscissa is the wavelength and the ordinate is the fluorescence intensity. Among them, Figure 1 A is the fluorescence emission spectrum of I1, Figure 1 B is the fluorescence emission spectrum of I2, Figure 1 C is the fluorescence emission spectrum of I3, Figure 1 D is the fluorescence emission spectrum of I4.

[0061] The results showed that the fluorescence intensities of the compounds I1, I2, I3 and I4 of the present invention at about 520 nm increased with the increase of the irradiation time, which proved that the compounds of the present invention could effectively generate reactive oxygen species after irradiation and had the potential for photodynamic therapy.

[0062] Example 6: Test on the ability of the carbazole / benzo[cd]indole compound of the present invention to generate superoxide anions

[0063] The ability of the compounds of the present invention to generate superoxide anions was detected by fluorescence spectroscopy, using dihydroethidium (DHE) as a scavenger for superoxide anions. The specific method was as follows: A solution of the compound of the present invention (10 μM) and the scavenger DHE (10 μM) was mixed, and then irradiated with a laser at 520 nm (50 mW / cm 2 ) for 3 min in total, and the test was carried out at intervals of 20 s. The fluorescence spectrometer was used to detect the change in the fluorescence emission spectrum at 607 nm of the compound of the present invention at different times after laser irradiation. Figure 2 Figure is the fluorescence emission spectrum diagram of the compounds I1, I2 and I3 of the present invention generating superoxide anions. The abscissa is the wavelength and the ordinate is the fluorescence intensity. Among them, Figure 2 A is the fluorescence emission spectrum diagram of I1, Figure 2 B is the fluorescence emission spectrum diagram of I2, Figure 2 C is the fluorescence emission spectrum diagram of I3, Figure 2 D is the fluorescence emission spectrum diagram of I4.

[0064] The results showed that the maximum fluorescence intensities of the compounds I1, I2, I3 and I4 increased with the increase of the irradiation time, proving that the compounds of the present invention can effectively generate a large amount of superoxide anions after irradiation with light and can play the role of type I photodynamic therapy.

[0065] Example 7: Test on the ability of the carbazole / benzo[cd]indole compound of the present invention to generate singlet oxygen

[0066] The ability of the compounds of the present invention to generate singlet oxygen was detected by fluorescence spectroscopy, using singlet oxygen fluorescence probe (SOSG) as a scavenger for singlet oxygen. The specific method was as follows: A solution of the compound of the present invention (5 μM) and the scavenger SOSG (5 μM) was mixed, and then irradiated with a laser at 520 nm (50 mW / cm 2 ) for 3 min in total, and the test was carried out at intervals of 20 s. The fluorescence spectrometer was used to detect the change in the fluorescence emission spectrum at 528 nm of the compound of the present invention at different times after laser irradiation. Figure 3 Figure is the ultraviolet absorption spectrum diagram of the compounds I1, I2, I3 and I4 of the present invention generating singlet oxygen. The abscissa is the wavelength and the ordinate is the absorbance value. Among them, Figure 3 A is the fluorescence emission spectrum diagram of I1, Figure 3 B is the fluorescence emission spectrum diagram of I2, Figure 3 C is the fluorescence emission spectrum diagram of I3, Figure 3 D is the fluorescence emission spectrum diagram of I4.

[0067] The results showed that no singlet oxygen generation was detected for Compound I1, while a small amount of singlet oxygen generation was detected for Compounds I2, I3, and I4. Therefore, it was proved that Compound I1 of the present invention had no significant singlet oxygen generation ability and could not significantly play the role of type II photodynamic therapy.

[0068] Example 8: Test on the ability of the carbazole / benzo[cd]indole compound of the present invention to generate hydroxyl radicals

[0069] The fluorescence spectrometry was used to detect the ability of the compound of the present invention to generate hydroxyl radicals, with hydroxyphenyl fluorescein (HPF) as the scavenger of hydroxyl radicals. The specific method was as follows: The solutions of the compound of the present invention (10 μM) and the scavenger HPF (10 μM) were mixed, and then irradiated with laser for a certain time. After the compound of the present invention was irradiated with a 520 nm (50 mW / cm 2 ) laser, the change in fluorescence intensity at 515 nm was detected. Figure 4 Figure 10 is the fluorescence emission spectrum diagram of Compounds I1, I2, I3, and I4 of the present invention generating hydroxyl radicals. The abscissa is the wavelength, and the ordinate is the fluorescence intensity. Among them, Figure 4 A is the fluorescence emission spectrum diagram of I1, Figure 4 B is the fluorescence emission spectrum diagram of I2, Figure 4 C is the fluorescence emission spectrum diagram of I3, Figure 4 D is the fluorescence emission spectrum diagram of I4.

[0070] The results showed that the maximum fluorescence intensity of Compound I1 increased significantly with the increase of irradiation time, and the growth level was significantly higher than that of I2, I3, and I4, indicating that Compound I1 of the present invention could generate a large amount of hydroxyl radicals after irradiation and could better play the role of type I photodynamic therapy.

[0071] Example 9: Activity test of the carbazole / benzo[cd]indole compound of the present invention in different tumor cells

[0072] The cytotoxicity and photodynamic therapy activity of the compound of the present invention against human colon cancer HT29 cell line, human lung cancer A549 cell line, and mouse breast cancer 4T1 cell line were evaluated by the tetramethyl thiazolyl blue colorimetric method (MTT) in vitro toxicity test. 4T1, A549, and HT29 cells were inoculated into 96-well plates at a density of 1×10 5 cells / mL, and 100 μL of medium containing 10% fetal bovine serum was added to each well. Under dark conditions, the compounds at test concentrations of 1, 2, 5, 10, and 20 μM were tested. Compounds I1, I2, and I3 were added to the medium and incubated for 24 hours. Under light conditions, the same concentration of compounds was used, and a 520 nm laser (100 mW / cm 2)Irradiate for 10 minutes, and then incubate for another 24 hours. Under hypoxic conditions (37 °C, 2% O2, 5% CO2), the cells are incubated in a three-gas incubator for 12 hours, then treated with compounds (1, 2, 5, 10, and 20 μM) for 4 hours, and then irradiated with a 520 nm laser (100 mW / cm 2 )for 10 minutes and continue to incubate for 24 hours. After removing the medium, add MTT solution (5 mg / mL, 20 μL) to each well and incubate at 37 °C for 4 hours. Discard the medium and add 150 μL of DMSO to each well to dissolve the formazan crystals. Measure the absorbance at a wavelength of 490 nm using a microplate reader to calculate the cell growth inhibition rate.

[0073] After a series of anti-proliferative activity tests on the compounds of the present invention against tumor cells, the experimental results show (Table 1) that it is found that under normoxic conditions (37 °C, 21% O2, 5% CO2), compounds I1, I2, I3, and I4 of the present invention have certain inhibitory activities against the proliferation of HT29, A549, and 4T1 tumor cells; under hypoxic conditions (37 °C, 2% O2, 5% CO2), compound I1 has certain inhibitory activities against the proliferation of HT29, A549, and 4T1 tumor cells, and the inhibitory activity against tumor cells is significantly enhanced after light irradiation, indicating that the compounds of the present invention have the characteristics of hypoxia-tolerant photodynamic therapy and play a significant chemo / photodynamic therapy role on tumor cells.

[0074] Table 1 Inhibitory activities of some compounds of the present invention against tumor cells under normoxic and hypoxic conditions

[0075]

[0076]

[0077] ND: Not tested.

[0078] Example 10: In vivo anti-tumor activity test of the carbazole / benzo[cd]indole compounds of the present invention

[0079] Select 6-8-week-old female Balb / c mice, 18-20 g, purchased from Nanjing Jicui Yakang Biotechnology Co., Ltd. The feeding conditions and experimental operations are all in strict accordance with the relevant requirements and standards of the Experimental Animal Management and Use Ethics Committee of Nantong University. Inject 100 μL of PBS cell suspension containing 2 × 10 6 4T1 breast cancer cells into the right axilla of the mice. When the tumor size reaches 80 mm 3 , randomly divide the mice into 4 groups: PBS, [Ru(bpy)3] 2+ (RuB)+L, I1, I1+L. Administer the drug by intratumoral injection at a dose of 5 mg / kg. After 4 hours of drug administration, irradiate with a 520 nm laser (200 mW / cm2 , (10 min), the tumor volume was monitored continuously for 15 days for 21 days. After the experiment, the mice were sacrificed, and the tumors were excised from each group for in vivo anti-tumor research. The formula for calculating the tumor volume is as follows: V = 1 / 2 × a × b 2 . V represents the tumor volume, a represents the longest diameter of the tumor site, and b represents the diameter perpendicular to a. Figure 5 are the test results of the in vivo anti-tumor activity of Compound I1 of the present invention. Among them, Figure 5 A is the tumor photos of different groups after the treatment, Figure 5 B is the growth change of the tumor volume of different groups.

[0080] The experimental results show that( Figure 5 ), after intratumoral injection and then 520 nm laser treatment, the inhibition rate of the RuB + L treatment group was only 51.27%, proving that the photodynamic therapy effect of RuB alone was poor. The inhibition rate of the I1 treatment group reached 75.82%, proving that I1 itself has good chemical activity. While the tumor growth of the mice in the I1 + L treatment group almost stagnated, and the inhibition rate compared to the PBS group reached 93.06%, indicating that I1 showed effective anti-tumor effects.

[0081] Example 11: In vitro photodynamic antibacterial ability test of the carbazole / benzo[cd]indole compound of the present invention

[0082] The broth microdilution method (turbidimetry) was used to evaluate the antibacterial activity against Staphylococcus aureus (S. aureus) and methicillin-resistant Staphylococcus aureus (MRSA). Nutrient broth medium (LB) was used to prepare the drug-containing suspension diluted twice. The specific method was as follows: In a 96-well plate, 100 μL of the diluted bacterial suspension and 100 μL of the compound solution of the present invention (10 μM) were added to each well, and the final volume in each well of the 96-well plate was controlled to 200 μL. It was irradiated with a 520 nm laser (100 mW / cm 2 ) for 10 minutes. After irradiation, the plate was incubated at 37 °C for 16 hours. By observing the turbidity and comparing it with the control, the absorbance value at 450 nm was measured, and the ratio to the blank group was used to indicate the bacterial survival rate.

[0083] Table 2 The effect of some compounds of the present invention on the survival rate of bacteria % (10 μM)

[0084]

[0085] ND: Not tested.

[0086] As shown in Table 2 above, the compounds of the present invention have antibacterial activities to varying degrees against Staphylococcus aureus and methicillin-resistant Staphylococcus aureus at a concentration of 10 μM, and the bacterial survival rate of the compounds of the present invention decreases significantly under light conditions, proving that the compounds of the present invention have a strong photodynamic antibacterial effect.

[0087] The above has described the embodiments of the present invention in detail, but the above content is only the preferred embodiment of the present invention and cannot be considered as limiting the scope of implementation of the present invention. All equivalent changes and improvements made according to the scope of the present invention application shall still fall within the scope covered by the patent of the present invention.

Claims

1. A carbazole / benzo[cd]indole compound, characterized in that, The carbazole / benzo[cd]indole compound has a structure shown in general formula I: Wherein, R is selected from a halogen atom or N(Ph)2.

2. The carbazole / benzo[cd]indole compound according to claim 1, wherein R is selected from Br or N(Ph)2.

3. The method for preparing a carbazole / benzo[cd]indole compound according to claim 1 or 2, characterized in that, R represents Br, and the synthetic route of the preparation method is as follows: The preparation method comprises the following steps: S1. N-ethylcarbazole-3-carboxaldehyde was brominated by NBS to obtain compound 3; S2. Compound 3 and 1,2-dimethyl-1H-benzo[cd]indol-1-ium undergo Knoevenagel condensation reaction in the presence of a catalytic amount of piperidine to obtain compound I3, which is the carbazole / benzo[cd]indole compound.

4. The method for preparing a carbazole / benzo[cd]indole compound according to claim 1 or 2, characterized in that, R represents (Ph)2N, and the synthetic route of the preparation method is as follows: S1. N-ethylcarbazole-3-carboxaldehyde was brominated by NBS to obtain compound 3; The synthetic route of step S1 is as follows: S2. Compound 3 and diphenylamine react in the presence of palladium acetate, tri-tert-butylphosphine, and cesium carbonate to obtain compound 4; The synthetic route of step S2 is as follows: S3. Compound 4 reacts with 1,2-dimethyl-1H-benzo[cd]indol-1-ium in the presence of a catalytic amount of piperidine to undergo a Knoevenagel condensation reaction to obtain compound I4, which is the carbazole / benzo[cd]indole compound; The synthetic route of step S3 is as follows:

5. Use of a carbazole / benz[cd]indole compound of formula I in the preparation of a drug or reagent for photodynamic therapy of a disease; Among them, R is selected from one of NO2, NH2, Br and N(Ph)2.

6. The application according to claim 5, wherein, The disease is a tumor, and the tumor is selected from colon cancer, lung cancer or breast cancer.

7. The application according to claim 5, wherein The disease is a disease associated with bacterial infection.

8. The application according to claim 7, wherein The bacteria is selected from Staphylococcus aureus or methicillin-resistant Staphylococcus aureus.

9. The application according to any one of claims 4-8, characterized in that, The photodynamic therapy is performed by a method comprising the following steps: S1. administering the carbazole / benzo[cd]indole compound to a subject; and S2. Irradiating the subject with light having a wavelength of 450 to 520 nm.

10. The use according to claim 9, characterized in that, The method has one or more of the following features: (1) In step S2, the irradiation time is 3 to 10 minutes; (2) In step S2, irradiate with a light intensity of 50-200 mW / cm 2 .

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