5, 9-di-tert-butyl-12-mesitylene naphtho indolizine phenothiazine compound as well as preparation method and application of 5, 9-di-tert-butyl-12-mesitylene naphtho indolizine phenothiazine compound

Abstract

The invention belongs to the technical field of organic synthesis and medicinal chemistry, and discloses efficient synthesis of a novel polycyclic aromatic hydrocarbon derivative 5, 9-di-tert-butyl-12-mesitylene naphtho indolizine phenothiazine and an anti-cancer application of the novel polycyclic aromatic hydrocarbon derivative 5, 9-di-tert-butyl-12-mesitylene naphtho indolizine phenothiazine. According to the compound, a mesitylene group is introduced for the first time on the basis of a naphthoindolizine phenothiazine skeleton. The synthesis conditions are mild, and the yield is as high as 91%. In-vitro anti-cancer activity research shows that the compound has remarkable broad-spectrum anti-cancer activity and shows strong proliferation inhibition on human cervical cancer Hela cells (IC50 = 1.6 mu M), human lung cancer A549 cells (IC50 = 13.29 mu M) and human prostatic cancer PC-3 cells (IC50 = 19.1 mu M). Particularly, the activity on Hela cells is obviously superior to that of a clinical first-line drug cis-platinum. Preliminary toxicity evaluation shows that the toxicity of the compound to human normal umbilical vein endothelial cells (HUVEC) is far lower than that of cancer cells, and the selectivity is good.

Description

Technical field:

[0001] The present invention belongs to the technical field of organic synthesis and medicinal chemistry, and particularly relates to a synthesis method of a 5,9-di-tert-butyl-12-mesityl naphtho-indolizine-phenothiazine compound and its anti-cancer application. Background technology:

[0002] Phenothiazine is a tricyclic heterocyclic compound composed of two benzene rings bridged by sulfur and nitrogen atoms. Since its discovery in the late 19th century, its unique molecular structure and modifiable properties have attracted continuous attention in the fields of medicinal chemistry, materials science, and industrial applications. Through systematic structural modification (such as the introduction of substituents and heterocyclic modification), phenothiazine derivatives have exhibited diverse biological activities and physicochemical properties, among which breakthrough applications in the pharmaceutical field are particularly prominent. In the field of neuropsychiatric treatment, phenothiazine derivatives, as the first generation of typical antipsychotic drugs (such as chlorpromazine and fluphenazine), have revolutionized the treatment strategy of schizophrenia through the dopamine receptor antagonist mechanism. Recent studies have shown that through precise structural modification, this type of compound can expand its multiple pharmacological activities such as antitumor, antiviral, antibacterial, anti-inflammatory and antioxidant. [Sarmiento.GP, Martini.MF, Vitale.RG, Fabian.LE, Afeltra.J., Vega.D., Moltrasio.GY, Moglioni.AGArab.J.Chem., 2017, 12, 21-32.]

[0003] Significant progress has been made in anti-tumor research. For example, the cyanoacrylamido phenothiazine derivatives designed by the Krishnan team showed excellent in vitro anti-proliferative effects on human pancreatic cancer cell lines AsPC1 and SW1990. [Krishnan.KG, Kumar.CU, Lim.W.-M., Mai.C.-W., Thanikachalam.PV, Ramalingan.CJMol.Struct., 2020, 1199, 127037.] Morak- The half inhibitory concentration (IC50) of the aza-phenothiazine derivatives developed by et al. on the breast cancer MCF-7 cell line 50 ) were <5 μg·mL -1 , which was significantly better than the cisplatin control (IC 50 =7.4 μg·mL -1 ); its IC against melanoma C-32 cell line 50 The values were 7.5 μg·mL -1 and 6.6 μg·mL -1 , showing antitumor activity comparable to cisplatin. [Morak- B., Pluta.K., Latocha.M., M.Med.Chem.Res., 2016, 25, 2425-2433.] These breakthrough results highlight the core value of the phenothiazine skeleton in the development of new anti-tumor drugs.

[0004] Current structural optimization strategies focus on two dimensions: (1) modification of nitrogen-sulfur heterocycles, including functionalization of N-substituents (alkyl / aryl) and regulation of the oxidation state of sulfur atoms (sulfoxide / sulfonation); and (2) modification of aromatic ring systems, including introduction of substituents at positions 1-4 and 6-9, or expansion of molecular diversity through strategies such as homo- / heteroaromatic ring substitution, construction of polyaromatic ring systems, and nitrogen-heterocycle fusion. It is worth noting that the research on novel derivatives based on the expansion of π-conjugated systems (e.g., mesityl-substituted phenothiazines) is still in the exploratory stage, which provides an innovative direction for the development of functional molecules with special optoelectronic properties or enhanced pharmacological activities. Summary of the invention:

[0005] Based on the above content, the present invention provides a 5,9-di-tert-butyl-12-mesityl naphtho-indolizine-phenothiazine compound, a synthesis method thereof, and an anti-cancer application thereof.

[0006] One of the technical solutions of the present invention is a 5,9-di-tert-butyl-12-mesityl naphthoindolizine phenothiazine compound, the structural formula of which is shown in formula (I):

[0007]

[0008] The second technical solution of the present invention is a method for preparing the above-mentioned 5,9-di-tert-butyl-12-mesityl naphthoindolizine-phenothiazine compound, comprising the following steps: using a 12-bromo-5,9-di-tert-butyl-naphthoindolizine-phenothiazine compound represented by formula (II) and mesityl boronic acid as starting materials, heating the mixture in a solvent in the presence of tris(dibenzylideneacetone)dipalladium, 2-dicyclohexylphosphine-2′,6′-dimethoxybiphenyl, and anhydrous potassium carbonate to obtain the compound;

[0009] Structural formula (II)

[0010]

[0011] Reaction equation:

[0012]

[0013] Furthermore, the molar ratio of the 12-bromo-5,9-di-tert-butylnaphthoindolizine and phenothiazine compound, mesityleneboric acid, tris(dibenzylideneacetone)dipalladium, 2-dicyclohexylphosphine-2′,6′-dimethoxybiphenyl, and anhydrous potassium carbonate is 1:3:0.2:0.4:3.

[0014] Furthermore, the concentration of the 12-bromo-5,9-di-tert-butylnaphthoindolizine-phenothiazine compound in the solvent is 0.02 mol / L, the solvent is toluene and water, the volume ratio of toluene and water is 10:1, the temperature is 100° C., and the reaction time is 24 h.

[0015] Furthermore, after the reaction is completed, the process further includes water washing, dichloromethane extraction, drying and filtration, concentration, and purification; the purification is performed by silica gel column chromatography (eluent is n-hexane: dichloromethane = 50 / 1, V / V).

[0016] The third technical solution of the present invention is the use of the above-mentioned 5,9-di-tert-butyl-12-mesityl naphtho-indolizine-phenothiazine compound in the preparation of anticancer drugs.

[0017] Furthermore, the anticancer drug is used to treat human cervical cancer (HeLa).

[0018] Furthermore, the anticancer drug is used to treat human non-small cell lung cancer (A549).

[0019] Furthermore, the anticancer drug is used to treat human prostate cancer (PC-3).

[0020] Compared with the prior art, the present invention has the following beneficial effects:

[0021] This invention, through precise molecular design, constructs the first mesitylene derivative based on the naphthoindole phenothiazine skeleton. This compound is efficiently synthesized via a Suzuki coupling reaction under mild conditions, with a yield of up to 91% for the target product. In vitro antitumor activity studies have shown that this compound exhibits broad-spectrum anticancer activity against human cervical cancer HeLa cells (IC 50 =1.6 μM), non-small cell lung cancer A549 cells (IC 50 =13.29 μM) and human prostate cancer PC-3 cells (IC 50 =19.1μM) showed significant proliferation inhibitory effects. Compared with the clinical first-line chemotherapy drug cisplatin, the compound has significantly improved inhibitory activity against human cervical cancer Hela cells and human non-small cell lung cancer A549 cells. Preliminary toxicity evaluation showed that the cytotoxicity of the compound to normal human umbilical vein endothelial cells (HUVEC) was significantly lower than that to cancer cells, showing a good selective inhibitory effect. This study not only provides an innovative strategy for the synthesis of new polycyclic aromatic hydrocarbon derivatives, but its significant anti-cancer effect also provides candidate compounds with important research value for the development of anti-cancer drugs. Description of the drawings:

[0022] Figure 1This is a hydrogen nuclear magnetic resonance spectrum of the 5,9-di-tert-butyl-12-mesityl naphthoindolizine phenothiazine compound prepared in Example 1 of the present invention;

[0023] Figure 2 This is the carbon NMR spectrum of the 5,9-di-tert-butyl-12-mesityl naphthoindolizine phenothiazine compound prepared in Example 1 of the present invention;

[0024] Figure 3 This is the single crystal diffraction pattern of the 5,9-di-tert-butyl-12-mesityl naphtho-indolizine-phenothiazine compound prepared in Example 1 of the present invention. Specific implementation method:

[0025] The present invention is described in detail below with reference to specific embodiments.

[0026] Example 1

[0027] Synthesis of 5,9-di-tert-butyl-12-mesityl-naphtho-indolizine-phenothiazine compounds

[0028] In a 120 mL pressure-resistant reaction flask, add 12-bromo-5,9-di-tert-butylnaphthoindolizine-phenothiazine (0.1 g, 0.2 mmol), mesitylboronic acid (0.1 g, 0.6 mmol), tris(dibenzylideneacetone)dipalladium (0.04 g, 0.04 mmol), 2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl (0.03 g, 0.08 mmol), and anhydrous potassium carbonate (0.08 g, 0.6 mmol). Under argon, add anhydrous toluene (10 mL) and water (1 mL), and continue bubbling for 3 minutes. Stir at 100°C for 24 hours. After the reaction, cool to room temperature, add dichloromethane (50 mL), wash with water three times, dry over anhydrous magnesium sulfate, filter, and concentrate the filtrate to dryness. The crude product was separated and purified by silica gel column chromatography (eluent: n-hexane:dichloromethane = 50 / 1, V / V) to give the desired product (0.1 g, yellow solid, yield 91%).

[0029] 1 H NMR (600MHz, C6D6, 298K) δ7.81 (d, J=7.2Hz, 1H), 7.78 (s, 2H), 7.75 (s, 1H), 7.57 (d, J=8.4Hz, 1H), 7 .44(t, J=7.8Hz, 1H), 7.14(s, 2H), 7.08(s, 2H), 2.37(s, 3H), 2.11(s, 6H), 1.35(s, 9H), 1.20(s, 9H); 13C NMR (150MHz, C6D6, 298K) δ147.6, 147.4, 137.9, 137.7, 136.9, 131.2, 130.4, 130.0, 128.8, 128.4, 128.3, 127.4, 127.1, 123.1, 122.8 [M + ]calcd.For C 39 H 37 NS, 551.2647; found, 551.2642.

[0030] Example 2

[0031] Synthesis of 5,9-di-tert-butyl-12-mesityl-naphtho-indolizine-phenothiazine compounds

[0032] In a 120 mL pressure-resistant reaction flask, 12-bromo-5,9-di-tert-butylnaphthoindolizine-phenothiazine (0.15 g, 0.3 mmol), mesitylboronic acid (0.15 g, 0.9 mmol), tris(dibenzylideneacetone)dipalladium (0.06 g, 0.06 mmol), 2-dicyclohexylphosphine-2′,6′-dimethoxybiphenyl (0.05 g, 0.12 mmol), and anhydrous potassium carbonate (0.12 g, 0.9 mmol) were added sequentially. Under argon protection, anhydrous toluene (15 mL) and water (1.5 mL) were added, and bubbling was continued for 3 minutes. The mixture was stirred at 100°C for 24 hours. After the reaction, the mixture was cooled to room temperature, and dichloromethane (50 mL) was added. The mixture was washed with water three times, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated to dryness. The crude product was separated and purified by silica gel column chromatography (eluent: n-hexane:dichloromethane = 50 / 1, V / V) to give the desired product (0.15 g, yellow solid, yield 91%).

[0033] 1 H NMR (600MHz, C6D6, 298K) δ7.81 (d, J=7.2Hz, 1H), 7.78 (s, 2H), 7.75 (s, 1H), 7-57 (d, J=8.4Hz, 1H), 7 .44(t, J=7.8Hz, 1H), 7.14(s, 2H), 7.08(s, 2H), 2.37(s, 3H), 2.11(s, 6H), 1.35(s, 9H), 1.20(s, 9H); 13C NMR (150MHz, C6D6, 298K) δ147.6, 147.4, 137.9, 137.7, 136.9, 131.2, 130.4, 130.0, 128.8, 128.4, 128.3, 127.4, 127.1, 123.1, 122.8 [M + ]calcd.For C 39 H 37 NS, 551.2647; found, 551.2642.

[0034] Example 3

[0035] Synthesis of 5,9-di-tert-butyl-12-mesityl-naphtho-indolizine-phenothiazine compounds

[0036] In a 120 mL pressure-resistant reaction flask, add 12-bromo-5,9-di-tert-butylnaphthoindolizine-phenothiazine (0.2 g, 0.4 mmol), mesitylboronic acid (0.2 g, 1.2 mmol), tris(dibenzylideneacetone)dipalladium (0.08 g, 0.08 mmol), 2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl (0.06 g, 0.16 mmol), and anhydrous potassium carbonate (0.16 g, 1.2 mmol). Under argon, add anhydrous toluene (20 mL) and water (2 mL), and continue bubbling for 3 minutes. Stir at 100°C for 24 hours. After the reaction, cool to room temperature, add dichloromethane (50 mL), wash with water three times, dry over anhydrous magnesium sulfate, filter, and concentrate the filtrate to dryness. The crude product was separated and purified by silica gel column chromatography (eluent: n-hexane:dichloromethane = 50 / 1, V / V) to give the desired product (0.2 g, yellow solid, yield 91%).

[0037] 1 H NMR (600MHz, C6D6, 298K) δ7.81 (d, J=7.2Hz, 1H), 7.78 (s, 2H), 7.75 (s, 1H), 7.57 (d, J=8.4Hz, 1H), 7 .44(t, J=7.8Hz, 1H), 7.14(s, 2H), 7.08(s, 2H), 2.37(s, 3H), 2.11(s, 6H), 1.35(s, 9H), 1.20(s, 9H); 13C NMR (150MHz, C6D6, 298K) δ147.6, 147.4, 137.9, 137.7, 136.9, 131.2, 130.4, 130.0, 128.8, 128.4, 128.3, 127.4, 127.1, 123.1, 122.8 [M + ]calcd.For C 39 H 37 NS, 551.2647; found, 551.2642.

[0038] Example 4

[0039] The anticancer activity of 5,9-di-tert-butyl-12-mesityl-naphthoindolizine phenothiazine compound against human cervical cancer cells (Hela), human non-small cell lung cancer cells (A549) and human prostate cancer cells (PC-3) was tested. The specific experimental methods are as follows:

[0040] The cytotoxicity of 5,9-di-tert-butyl-12-mesityl-1-methylphenylnaphthoquinone-indolizine-phenothiazine compounds to human cervical cancer cells (Hela), human non-small cell lung cancer cells (A549), human prostate cancer cells (PC-3) and normal human umbilical vein endothelial cells (HUVEC) was studied by MTT thiazolyl blue colorimetry. 5 A single cell suspension of 100 μL / mL was inoculated on a 96-well culture plate, with 100 μL per well. After culturing for 24 h in a 37°C incubator with a volume fraction of 5% carbon dioxide and saturated humidity, 100 μL of the sample to be tested at different concentrations was added to the culture plate, with 6 replicates for each concentration. After culturing for 48 h, 100 μL (1 mg / mL) of MTT staining solution was added to each well, and the plate was incubated for another 4 h in the incubator. The supernatant was removed, and then 200 μL of the sample to be tested was added to each well. DMSO was fully shaken for 15 minutes, and the OD sample value was measured at a wavelength of 595 nm on a microplate reader. The blank group of the experiment was 100 μL of serum-free DMEM culture medium with DMSO reagent added instead of the sample. The absorbance value at this time was the OD blank value. The inhibition rate of 5,9-di-tert-butyl-12-mesitylphenylnaphthoquinone-indolizine-phenothiazine compound on Hela, A549, PC-3, and HUVEC cells was calculated by formula (1). Finally, the IC 50 .

[0041] m=1-n=1-OD样 / OD 空白 (1)

[0042] m: inhibition rate n: cell survival rate

[0043] The results of the anticancer activity test of 5,9-di-tert-butyl-12-mesityl naphtho-indolizine-phenothiazine compound against Hela, A549 and PC-3 are shown in Table 1:

[0044] Table 1. Cytotoxicity of 5,9-di-tert-butyl-12-mesityl-1-phenyl-naphthoquinone-indolizine-phenothiazine compounds and positive control drug cisplatin (DDP) to Hela, A549, PC-3, HUVEC (data in the table are IC 50 value):

[0045] Compound Hela A549 PC-3 HUVEC This patent 1.6 13.29 19.1 157.28 DDP 18.58 51.92 14.51 26.48

[0046] As shown in Table 1, the 5,9-di-tert-butyl-12-mesityl-1,2-trimethylphenylnaphthoquinone-indolizine-phenothiazine compounds Hela, A549, and PC-3 synthesized in this patent exhibited varying degrees of inhibitory effects with significant effects. Among them, the inhibitory effects of Hela and A549 were significantly better than those of the positive control drug cisplatin (DDP), and the toxicity was relatively low, indicating that they have the potential to be developed into anti-cancer drugs.

Claims

1. A 5,9-di-tert-butyl-12-mesityl naphthoindolizine phenothiazine compound, characterized in that: The structural formula is shown in formula (I):

2. The method for preparing the 5,9-di-tert-butyl-12-mesityl naphtho-indolizine phenothiazine compound according to claim 1, wherein: The following steps are involved: The 12-bromo-5,9-di-tert-butylnaphthoindolizine-phenothiazine compound represented by formula (II) and mesityleneboronic acid are used as starting materials, and a heating reaction is carried out in a solvent in the presence of tris(dibenzylideneacetone)dipalladium, 2-dicyclohexylphosphine-2',6'-dimethoxybiphenyl and anhydrous potassium carbonate to obtain the product. Structural formula (II) 3. The method for preparing the 5,9-di-tert-butyl-12-mesityl naphtho-indolizine phenothiazine compound according to claim 2, wherein: The molar ratio of the 12-bromo-5,9-di-tert-butylnaphthoindolizine and phenothiazine compound, mesityleneboric acid, tris(dibenzylideneacetone)dipalladium, 2-dicyclohexylphosphine-2',6'-dimethoxybiphenyl and anhydrous potassium carbonate is 1:3:0.2:0.4:

3.

4. The method for preparing the 5,9-di-tert-butyl-12-mesityl naphtho-indolizine phenothiazine compound according to claim 2, wherein: The concentration of the 12-bromo-5,9-di-tert-butylnaphthoindolizine-phenothiazine compound in the solvent is 0.02 mol / L, the solvent is toluene and water, the volume ratio of toluene to water is 10:1, the temperature is 100° C., and the reaction time is 24 h.

5. Use of the 5,9-di-tert-butyl-12-mesityl naphthoindolizine phenothiazine compound according to claim 1 in the preparation of anticancer drugs.

6. Use of the 5,9-di-tert-butyl-12-mesityl naphthoindolizine phenothiazine compound according to claim 5 in the preparation of anticancer drugs, characterized in that: The anticancer drug is used for treating human cervical cancer (Hela).

7. Use of the 5,9-di-tert-butyl-12-mesityl naphthoindolizine phenothiazine compound according to claim 5 in the preparation of anticancer drugs, characterized in that: The anticancer drug is used to treat human non-small cell lung cancer (A549).

8. Use of the 5,9-di-tert-butyl-12-mesityl naphthoindolizine phenothiazine compound according to claim 5 in the preparation of anticancer drugs, characterized in that: The anticancer drug is used for treating human prostate cancer (PC-3).

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