A carbazole derivative and its preparation method and application

Abstract

The present invention belongs to the field of medicinal chemistry and pharmacology, and discloses a carbazole derivative, its preparation method, and application. The carbazole derivative has the structure represented by formula (I). A compound is obtained by dissolving cedrin 1 and ferric acetylacetonate in ethanol:ethylene glycol, adding phenylsilane, and reacting the resulting compound. The compound is then refluxed in ethanol with a phenylhydrazine derivative under the catalysis of p-toluenesulfonic acid to obtain the carbazole derivative. These carbazole derivatives have been found to exhibit strong tumor cell growth inhibition activity and are expected to be used as anti-tumor drugs.

Description

Technical Field

[0001] The invention belongs to the technical field of medicinal chemistry and pharmacology, and relates to a carbazole derivative and a preparation method and application thereof. Background Art

[0002] The development of medicinal plants has become a hot research topic in the medical field. Over the past few decades, natural products have garnered increasing attention due to their excellent therapeutic effects and low toxicity. The natural bicyclic conjugated sesquiterpenoid compound, bucubitrone, was initially considered an insect repellent with insecticide activity against ticks, mosquitoes, and termites, as well as for the treatment of human-related diseases such as chikungunya, dengue fever, and Zika virus. Recent research data indicates that bucubitrone exhibits a wide range of pharmacological activities, including anti-inflammatory, antibacterial, anticancer, neuroprotective, cardioprotective, and hepatoprotective effects. As a chemical drug backbone, the carbazole structure possesses a wide range of biological activities and holds a place in drugs in various therapeutic areas, including antihypertensive, antiproliferative, antiviral, antitumor, analgesic, anti-inflammatory, and antibacterial. The most important therapeutic area for carbazole structures is anti-tumor, and several carbazole alkaloids, such as staurosporine, have been reported to possess significant antitumor activity. Summary of the Invention

[0003] In view of this, the present invention aims to reshape the skeleton of cedarone and obtain a batch of new-structure carbazole derivatives through chemical transformation, which have strong tumor cell proliferation inhibition activity.

[0004] The present invention provides a carbazole derivative having a structure shown in formula (I):

[0005]

[0006] Here, R represents one of H, 2-CH3, 4-OCH3, 4-F, 4-Cl, and 4-Br.

[0007] Furthermore, the carbazole derivative has a structure as shown in any one of Formulas 3a-3d:

[0008]

[0009] in,

[0010] When R is H, the carbazole derivative is a compound having a structure shown in Formula 3a;

[0011] When R is 2-CH3, the carbazole derivative is a compound having a structure shown in formula 3b;

[0012] When R is 4-OCH3, the carbazole derivative is a compound having a structure shown in formula 3c;

[0013] When R is 4-F, the carbazole derivative is a compound having a structure shown in Formula 3d.

[0014] The present invention also provides a method for preparing a carbazole derivative, comprising the following steps:

[0015] Dissolve cedrin 1 and ferric acetylacetonate in ethanol:ethylene glycol, add phenylsilane, and react to obtain compound 2. Compound 2 is then refluxed in ethanol with a phenylhydrazine derivative under the catalysis of p-toluenesulfonic acid to obtain a carbazole derivative 3.

[0016] Wherein, the reaction formula of the reaction is:

[0017]

[0018] wherein R represents one of H, 2-CH3, 4-OCH3, 4-F, 4-Cl, and 4-Br.

[0019] Specifically, the preparation method comprises the following steps:

[0020] (1) Dissolve cedrin 1 and ferric acetylacetonate in a mixed solvent of ethanol and ethylene glycol, then add phenylsilane and react at 50-70° C. to obtain a first reaction solution. The first reaction solution is cooled to room temperature, quenched with water, washed with saturated brine, extracted with ethyl acetate, and the organic phase is collected and concentrated under reduced pressure to obtain a red oil. Flash column chromatography is used to obtain a colorless oil, namely compound 2, wherein the molar ratio of cedrin 1, ferric acetylacetonate and phenylsilane is 1:0.3:1.5;

[0021] (2) Compound 2 is dissolved in anhydrous ethanol, and a phenylhydrazine derivative and p-toluenesulfonic acid are added, and the mixture is refluxed to obtain a second reaction solution. After the second reaction solution is cooled, saturated sodium bicarbonate is added to quench the reaction, and the mixture is extracted with ethyl acetate. The organic phase is collected and washed with water, saturated brine, dried over anhydrous MgSO4, concentrated under reduced pressure, and then flash column chromatography is performed to obtain a carbazole derivative, wherein the molar ratio of compound 2, phenylhydrazine derivative, and p-toluenesulfonic acid is 1:1.5:1.5;

[0022] Wherein, the phenylhydrazine derivative is one of phenylhydrazine, 2-methylphenylhydrazine, 4-methoxyphenylhydrazine, 4-fluorophenylhydrazine, 4-chlorophenylhydrazine and 4-bromophenylhydrazine.

[0023] Furthermore, in step (1) of the above preparation method, the volume ratio of ethanol to ethylene glycol in the mixed solvent of ethanol and ethylene glycol is 5:1.

[0024] Furthermore, in step (1) of the above preparation method, the reaction temperature is 60° C. and the reaction time is 1 h.

[0025] Furthermore, in step (2) of the above preparation method, the reflux reaction temperature is 105° C. and the reaction time is 1 h.

[0026] The present invention also provides a use of the carbazole derivative in preparing a drug for preventing and treating cancer, wherein the cancer is one of gastric cancer, ovarian cancer, lung cancer and liver cancer.

[0027] Compared with the prior art, the present invention has the following technical effects:

[0028] (1) The carbazole derivatives provided by the present invention have a novel chemical skeleton, and in vitro tumor cell proliferation inhibition experiments have shown that these compounds have strong tumor cell proliferation inhibition activity, which is comparable to or superior to that of the positive control drug etoposide. They can be used in the preparation of drugs for preventing and treating gastric cancer, ovarian cancer, lung cancer, and liver cancer.

[0029] (2) In the prior art, the focus of structural research on natural products is more on biological transformation rather than chemical transformation. This invention proposes a ring distortion strategy to reshape the skeleton of cedarone and obtain a group of carbazole derivatives with new structures through chemical transformation. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for describing the embodiments or the prior art.

[0031] Figure 1 The NMR of compound 2 provided in Example 1 of the present invention is 1 H spectrum;

[0032] Figure 2 The NMR of compound 2 provided in Example 1 of the present invention is 13 C spectrum;

[0033] Figure 3 The nuclear magnetic resonance of the carbazole derivative (3a) provided in Example 2 of the present invention 1 H spectrum;

[0034] Figure 4 The nuclear magnetic resonance of the carbazole derivative (3a) provided in Example 2 of the present invention 13 C spectrum;

[0035] Figure 5 The COSY NMR spectrum of the carbazole derivative (3a) provided in Example 2 of the present invention;

[0036] Figure 6 The nuclear magnetic resonance of the carbazole derivative (3d) provided in Example 5 of the present invention 1 H spectrum;

[0037] Figure 7 The nuclear magnetic resonance of the carbazole derivative (3d) provided in Example 5 of the present invention 13 C spectrum. DETAILED DESCRIPTION

[0038] The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts are within the scope of protection of the present invention.

[0039] Example 1

[0040] 218 mg (1.0 mmol) of cedarone 1 was dissolved in a mixed solvent of ethanol and ethylene glycol (5 mL, ethanol:ethylene glycol volume ratio of 5:1). 106 mg (0.3 mmol) of ferric acetylacetonate and 162 mg (1.5 mmol) of phenylsilane were added and reacted at 60°C for 1 hour. The reaction solution was cooled to room temperature, quenched with water, washed with saturated brine, and extracted three times with ethyl acetate. The organic phase was collected and concentrated under reduced pressure to obtain a red residue. Flash column chromatography (petroleum ether:ethyl acetate = 150:1) gave 108 mg of a colorless oily liquid, i.e., compound 2 (yield 91%).

[0041] 1H NMR(400MHz,Chloroform-d)δ2.39(dqd,J=13.3,6.7,4.4Hz,1H,H-1),2.30–2.24(m,1H),2.2 0(s,1H),2.17–2.11(m,1H),2.05(ddd,J=14.4,4.5,1.7Hz,1H),1.90(dt,J=12.6,3.5Hz,1H) ,1.78–1.73(m,1H),1.72–1.69(m,2H),1.41(ddd,J=12.8,11.2,4.9Hz,1H),1.24–1.13(m,1H ),1.07(d,J=12.7Hz,1H),1.03(s,6H,CH3),0.92(s,3H,CH3),0.87(dd,J=6.7Hz,3H,1-CH3). 13 C NMR (100MHz, CDCl3) δ213.3,55.0,50.0,46.3,45.2,43.4,42.3,41.4,38.9,32.6,26.5,23.5,22.5,18.1,16.6.

[0042] Example 2

[0043] 216 mg (1.0 mmol) of compound 2 was dissolved in anhydrous ethanol (5 ml), followed by the addition of 164 mg (1.5 mmol) of phenylhydrazine and 258 mg (1.5 mmol) of p-toluenesulfonic acid. The mixture was refluxed at 105°C for 1 hour. After cooling, the reaction mixture was quenched by the addition of saturated sodium bicarbonate and extracted with ethyl acetate. The organic phase was collected and washed sequentially with water and saturated brine, dried over anhydrous MgSO₄, and concentrated under reduced pressure. Flash column chromatography (petroleum ether:ethyl acetate = 150:1) afforded the carbazole derivative 3a as a white solid (yield 53%).

[0044] 1 H NMR(400MHz,Chloroform-d)δ7.76(d,J=8.0Hz,1H,Ar-H),7.72(s,1H,NH),7.26(d,J=8.0Hz,1H,Ar-H),7.12–7.05 (m,1H,Ar-H),7.05–6.98(m,1H,Ar-H),2.66(ddd,J=13.5,11.5,5.7Hz,1H),2.53(dd,J=14.2,3.9Hz,2H),2.51–2.4 4(m,1H),2.38(dd,J=14.2,10.4Hz,1H),2.03(dt,J=12.1,3.6Hz,1H),1.96(ddt,J=11.6,5.5,3.2Hz,1H),1.81–1.7 3(m,2H),1.40–1.31(m,1H),1.35(s,3H,CH3),1.10(d,J=12.3Hz,1H),0.98(d,J=6.8Hz,6H,CH3),0.88(s,3H,CH3). 13 C NMR (100MHz, CDCl3) δ136.3,135.8,128.0,121.7,120.4,118.5,110.5,109.9,54.9,5 2.2,46.5,44.4,42.9,35.0,29.2,28.3,27.3,25.6,23.6,17.9,15.7.HRMS(ESI):m / z calcd for C 21 H 28 N:294.2222; found:294.2230[M+H] + .

[0045] Examples 3-5

[0046] According to the method of Example 2 above, compounds 3b-3d were prepared by replacing phenylhydrazine with 2-methylphenylhydrazine, 4-methoxyphenylhydrazine, and 4-fluorophenylhydrazine, respectively.

[0047] The following are the physicochemical data of compounds 3b-3d:

[0048] 3b: Yield 45%. 1 H NMR(400MHz,Chloroform-d)δ7.70(s,1H,NH),7.62(d,J=8.0Hz,1H,Ar-H),6.95(t,J=7.5Hz,1H,Ar-H),6.90( d,J=7.1Hz,1H),2.66(ddd,J=13.3,11.4,5.7Hz,1H),2.58(dd,J=14.8,4.1Hz,1H),2.54–2.46(m,1H),2.45(s ,3H,Ar-CH3),2.44–2.37(m,1H),2.03(dt,J=12.4,3.6Hz,1H),1.96(ddt,J=11.6,5.6,3.1Hz,1H),1.82–1.71 (m,1H),1.35(s,3H,CH3),1.40–1.30(m,1H),1.10(d,J=12.2Hz,1H),1.02–0.95(m,6H,CH3),0.89(s,3H,CH3). 13 C NMR (100MHz, CDCl3) δ135.7,135.5,127.4,121.2,119.5,119.5,118.7,110.5,54.9,52. 2,46.5,44.4,42.9,35.0,29.3,28.3,27.3,25.6,23.6,17.9,16.7,15.7.HRMS(ESI):m / z calcd for C 21 H 30 N:308.2378; found:308.2377[M+H] + .

[0049] 3c: Yield 31%. 11H NMR (400 MHz, Chloroform-d) δ 7.58 (s, 1H, NH), 7.21–7.17 (m, 1H, Ar-H), 7.09 (d, J = 8.7 Hz, 1H, Ar-H), 6.68 (dd, J = 8.7, 2.5 Hz, 1H, Ar-H), 3.76 (s, 3H, Ar-OCH3), 2.56 (ddd, J = 13.5, 11.6, 5.8 Hz, 1H), 2.48–2.43 (m, 1H), 2.44–2.33 (m, 1H), 2.30 (dd, J = 14.2, 10.4 Hz, 1H), 1.95 (dt, J = 12.0, 3.5 Hz, 1H), 1.89 (ddd, J = 11.6, 5.3, 2.6 Hz, 1H), 1.74–1.64 (m, 2H), 1.29 (s, 3H, CH3), 1.33–1.23 (m, 1H), 1.03 (d, J = 12.3 Hz, 1H), 0.90 (d, J = 5.9 Hz, 6H, CH3), 0.82 (s, 3H, CH3). 13 13C NMR (100 MHz, CDCl3) δ 153.1, 136.9, 131.6, 128.4, 110.7, 109.8, 109.4, 105.2, 56.2, 54.8, 52.3, 46.5, 44.4, 42.8, 35.0, 29.3, 27.9, 27.2, 25.7, 23.6, 17.9, 来 自 15.7. HRMS (ESI): m / z calcd for C 22 1 30 17H22NO: 324.2327; found: 324.2334 [M+H]+ + .

[0050] 3d: The yield was 43%. 1 It should be noted that there seems to be an incorrect expression "来 自" in the translation of line , which should be corrected in a more accurate translation context.H NMR(400MHz,)δ7.65(s,1H,NH),7.32(dd,J=11.0,2.5Hz,1H,Ar-H),7.07(dd,J=8.7,4.7Hz,1H,Ar-H),6. 75(td,J=9.0,2.5Hz,1H,Ar-H),2.53–2.47(m,1H),2.47–2.42(m,1H),2.41–2.34(m,1H),2.31(dd,J=14. 4,10.7Hz,1H),1.98–1.92(m,1H),1.88(ddt,J=11.6,5.8,3.3Hz,1H),1.73–1.69(m,1H),1.69-1.63(m,1 H),1.36-1.27(m,1H),1.26(s,3H,CH3),1.03(d,J=12.3Hz,1H),0.92–0.88(m,6H,CH3),0.81(s,3H,CH3). 13 C NMR (100MHz, CDCl3) δ157.2 (d, J = 232.0Hz), 137.8, 132.8, 128.2 (d, J = 9.6Hz), 110.62 (d, J = 9.9Hz), 110.36 (d, J = 4.3Hz), 106.75 (d,J=24.2Hz),106.9,106.6,54.7,52.3,46.5,44.4,42.8,34.9,29.3,28.0,27.1,25.6,23.6,17.9,15.7.HRMS(ESI):m / zcalcd for C 21 H 27 NF:312.2128;found:312.2130[M+H] + .

[0051] To better understand the essence of the present invention, the following pharmacological experimental results demonstrating the inhibitory effects of the carbazole derivatives provided herein on the growth of four tumor cell lines illustrate their novel applications in anti-tumor drug research. The pharmacological examples provide partial activity data for representative compounds. It should be noted that the pharmacological examples are intended to illustrate the present invention and are not intended to limit it. Simple modifications to the present invention based on its essence fall within the scope of the present invention.

[0052] Drug Experiment Example 1: Cytotoxicity Test of Compounds 3a-3d and Etoposide (VP-16) against Human Gastric Cancer Cells (MGC803)

[0053] Human gastric cancer cells (MGC803) were cultured in RPMI1640 medium supplemented with 10% fetal bovine serum, 100 U / mL penicillin, and 100 U / mL streptomycin. Cells were plated at a concentration of 5 × 103 cells per well in a 96-well plate and incubated at 37°C in a humidified atmosphere containing 5% CO2 for 24 hours.

[0054] Compounds 3a-3d were dissolved in DMSO and prepared into 1×10 -2 mol / L stock solution, the stock solution was diluted to the corresponding concentration with complete medium. Cells in logarithmic growth phase were seeded in 96-well plates. After 24 hours of attachment, different concentrations of compound solutions were added. Four parallel wells were set for each concentration. After 68 hours of culture, tetramethylthiazolium (MTT) solution was added. The culture was continued for 4 hours, the culture medium was discarded, 150 μL of dimethyl sulfoxide was added, and the cells were shaken for 10 minutes. The absorbance (A) value at 570 nm was measured with a microplate reader, and the half-maximal inhibitory concentration (IC) was calculated. 50 ), as shown in Table 1. According to Table 1, the IC 50 The positive control etoposide has an IC of 3.3 μM for MGC803 cells. 50 It is 4.6μM.

[0055] Drug Experimental Examples 2-4: Cytotoxic activity test of compounds 3a-3d and etoposide against human ovarian adenocarcinoma cells (SK-OV-3), human lung adenocarcinoma cells (A549), and human liver cancer cells (HepG2).

[0056] The method shown in Drug Experiment Example 1 was used to conduct pharmacological experiments on the growth inhibition of human gastric cancer cells (MGC803), human ovarian adenocarcinoma cells (SK-OV-3), human lung adenocarcinoma cells (A549), and human liver cancer cells (HepG2). The half-maximal inhibitory concentration (IC 50 ), as shown in Table 1.

[0057] Table 1 Cytotoxic activity test results of compounds 3a-3d and etoposide

[0058]

[0059]

[0060] As shown in Table 1, the carbazole derivatives provided by the present invention have significant biological activity. In vitro cytotoxicity tests on four types of tumor cells, namely human gastric cancer cells (MGC803), human ovarian adenocarcinoma cells (SK-OV-3), human lung adenocarcinoma cells (A549), and human liver cancer cells (HepG2), indicate that these carbazole derivatives having the structure represented by formula (1) have an inhibitory effect on tumor cell growth and may be developed into new anti-tumor drugs. From the above pharmacological examples, it can be seen that these compounds exhibit strong cytotoxic activity against all four types of tumor cells, with cytotoxic activity exceeding or comparable to that of the positive control, etoposide, indicating that they have the potential to be developed into anti-tumor drugs.

[0061] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A carbazole derivative, characterized in that The carbazole derivative has the structure shown in the following formula: wherein R represents one of H, 2-CH3, 4-OCH3, 4-F, 4-Cl, and 4-Br.

2. The carbazole derivative according to claim 1, characterized in that The carbazole derivative has a structure as shown in any one of Formulas 3a to 3d:

3. A method for preparing the carbazole derivative according to claim 1, characterized in that: The reaction formula of the preparation method is shown below: The preparation method comprises the following steps: S1. The cedar ketone 1 and iron acetylacetonate were dissolved in a mixed solvent of ethanol and ethylene glycol, and then phenylsilane was added and reacted at 50-70 ° C to obtain a first reaction solution; wherein the molar ratio of cedar ketone 1, iron acetylacetonate and phenylsilane was 1:0.3:1.5; S2. The first reaction solution was cooled to room temperature, the reaction was quenched with water, washed with saturated brine, extracted with ethyl acetate, and the organic phase was collected and concentrated under reduced pressure to obtain a red oil, which was then purified by flash column chromatography to obtain a colorless oil, which was compound 2; S3. Compound 2 was dissolved in anhydrous ethanol, and a phenylhydrazine derivative and p-toluenesulfonic acid were added, and the reaction was refluxed to obtain a second reaction solution; S4. After the second reaction solution was cooled, saturated sodium bicarbonate was added to quench the reaction, extracted with ethyl acetate, and the organic phase was collected and washed with water, saturated brine, dried over anhydrous MgSO4, concentrated under reduced pressure, and then flash column chromatography to obtain a carbazole derivative; the molar ratio of compound 2, phenylhydrazine derivative and p-toluenesulfonic acid was 1:1.5:1.5; Wherein, the phenylhydrazine derivative is one of phenylhydrazine, 2-methylphenylhydrazine, 4-methoxyphenylhydrazine, 4-fluorophenylhydrazine, 4-chlorophenylhydrazine and 4-bromophenylhydrazine.

4. The preparation method according to claim 3, characterized in that In step S1, in the mixed solvent of ethanol and ethylene glycol, the volume ratio of ethanol to ethylene glycol is 5:

1.

5. The preparation method according to claim 3, characterized in that In step S1, the reaction temperature is 60° C. and the reaction time is 1 h.

6. The preparation method according to claim 3, characterized in that In step S3, the temperature of the reflux reaction is 105° C., and the reaction time is 1 h.

7. Use of the carbazole derivative according to claim 1 in the preparation of a drug for preventing and treating cancer, wherein the cancer is one of gastric cancer, ovarian cancer, lung cancer and liver cancer.

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