Carbazole derivative as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of medicinal chemistry and pharmacology, and discloses a carbazole derivative as well as a preparation method and application thereof. The carbazole derivative has a structure as shown in a formula (I), and is prepared by the following steps: dissolving cacumen platycladi 1 and ferric acetylacetonate in ethanol and ethylene glycol, adding phenylsilane, reacting to obtain a compound, and then refluxing the compound and a phenylhydrazine derivative in ethanol under the catalysis of p-toluenesulfonic acid to obtain the carbazole derivative. The carbazole derivatives are found to have relatively strong tumor cell growth inhibition activity, and can be expected to be used as antitumor drugs. # imgabs0 #

Description

Technical Field

[0001] The invention belongs to the technical field of pharmaceutical 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 direction in the field of medicine. In the past few decades, natural products have received increasing attention due to their excellent therapeutic effects and low toxicity. The natural bicyclic conjugated sesquiterpenoid compound, bucatinone, was originally considered to be an insect repellent with insecticidal activity against ticks, mosquitoes and termites, as well as for the treatment of human-related diseases such as chikungunya, dengue fever and Zika virus. The latest research data results show that bucatinone has a wide range of pharmacological activities, including anti-inflammatory, antibacterial, anticancer, neuroprotective, myocardial protective, and liver protective effects. As a structural skeleton of chemical drugs, the carbazole structure has a wide range of biological activities and occupies a place in drugs in multiple therapeutic fields such as antihypertension, antiproliferation, antiviral, antitumor, analgesic, anti-inflammatory, and antibacterial. The most important therapeutic field of the carbazole structure is the anti-tumor field. It has been reported that a variety of carbazole alkaloids such as staurosporine have important antitumor activities. Summary of the invention

[0003] In view of this, the purpose of the present invention is to reshape the skeleton of cedar ketone and obtain a batch of carbazole derivatives with new structures through chemical transformation, and the derivatives have strong tumor cell proliferation inhibition activity.

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

[0005]

[0006] Wherein, 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 with 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) dissolving cedrin 1 and ferric acetylacetonate in a mixed solvent of ethanol and ethylene glycol, adding phenylsilane, and reacting 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 oily substance, which is then flash column chromatographed to obtain a colorless oily substance, i.e., 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 saline, 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, in the mixed solvent of ethanol and ethylene glycol, the volume ratio of ethanol to 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 an application 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 new chemical skeleton, and the in vitro tumor cell proliferation inhibition experiment found that such compounds have strong tumor cell proliferation inhibition activity, which is equivalent to or better than the positive control drug etoposide, and 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 process of studying the structure of natural products, the prior art focuses more on biological transformation rather than chemical transformation. The present invention proposes to use a ring distortion strategy to reshape the skeleton of cedar ketone and obtain a batch 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 drawings required for use in the embodiments or the description of the prior art are briefly introduced below.

[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 nuclear magnetic resonance spectrum of the carbazole derivative (3a) provided in Example 2 of the present invention;

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

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

[0038] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0039] Example 1

[0040] 218 mg (1.0 mmol) of chalcogenol 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 the mixture was reacted at 60°C for 1 hour. The reaction solution was cooled to room temperature, quenched with water, washed with saturated brine, extracted with ethyl acetate 3 times, the organic phase was collected, and concentrated under reduced pressure to obtain a red residue, which was flash column chromatography (petroleum ether: ethyl acetate = 150: 1) to obtain 108 mg of a colorless oily liquid, namely 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), and then 164 mg (1.5 mmol) of phenylhydrazine and 258 mg (1.5 mmol) of p-toluenesulfonic acid were added, and the mixture was refluxed at 105° C. for 1 hour. After the reaction solution was cooled, saturated sodium bicarbonate was added to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phase was collected and washed with water, saturated brine, dried over anhydrous MgSO4, and concentrated under reduced pressure. A white solid, i.e., carbazole derivative 3a (yield 53%), was obtained by flash column chromatography (petroleum ether: ethyl acetate = 150:1).

[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] Embodiment 3-5

[0046] According to the method of Example 2 above, phenylhydrazine was replaced by 2-methylphenylhydrazine, 4-methoxyphenylhydrazine and 4-fluorophenylhydrazine, respectively, to prepare compounds 3b-3d.

[0047] Listed below 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, 15.7. HRMS (ESI): m / z calcd for C 22 1 30 17H24NO: 324.2327; found: 324.2334 [M+H]+ + .

[0050] 3d: The yield was 43%. 1H 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] In order to better understand the essence of the present invention, the pharmacological experimental results of the inhibitory effect of the carbazole derivatives provided by the present invention on the growth of four tumor cell lines are used below to illustrate its new use in the field of anti-tumor drug research. The pharmacological examples provide partial activity data of representative compounds. It must be noted that the pharmacological examples of the present invention are used to illustrate the present invention rather than to limit the present invention. Simple improvements made to the present invention according to the essence of the present invention all belong to the scope of protection claimed by the present invention.

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

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

[0054] Compounds 3a-3d were dissolved in DMSO and 1×10 -2 mol / L stock solution, dilute the stock solution to the corresponding concentration with complete medium, take cells in logarithmic growth phase and inoculate them in 96-well plates, add compound solutions of different concentrations after 24 hours of attachment, set 4 parallel wells for each concentration, add tetramethylthiazolium (MTT) solution after 68 hours of culture, continue to culture for 4 hours, discard the culture medium, add 150μL of dimethyl sulfoxide, shake for 10 minutes, measure the absorbance (A) value at 570nm with an enzyme reader, and calculate the half inhibitory concentration (IC 50 ), as shown in Table 1. According to Table 1, the IC 50 The IC value of positive control etoposide for MGC803 cells was 3.3 μM. 50 It is 4.6μM.

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

[0056] Using the method shown in Drug Experiment Example 1, pharmacological experiments were performed 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), and the half 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 important biological activities. In vitro cytotoxic activity 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) show that the carbazole derivatives of the structure shown in formula (1) have an inhibitory effect on the growth of tumor cells and may be developed into new tumor prevention and treatment drugs. From the above pharmacological examples, we can see that these compounds show strong cytotoxic activity on these four types of tumor cells, and the cytotoxic activity exceeds or is equivalent to that of the positive control etoposide, and 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 principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A carbazole derivative, characterized in that: The carbazole derivative has a structure shown in the following formula: wherein R represents one of H, 3-CH3, 5-OCH3, 5-F, 5-Cl and 5-Br.

2. The carbazole derivative according to claim 1, characterized in that The carbazole derivative has a structure as shown in any one of Formulae 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 ferric acetylacetonate are dissolved in a mixed solvent of ethanol and ethylene glycol, and then phenylsilane is added and reacted at 50-70 ° C to obtain a first reaction solution; wherein the molar ratio of cedar ketone 1, ferric acetylacetonate and phenylsilane is 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, the organic phase was collected and concentrated under reduced pressure to obtain a red oil, and a colorless oil was obtained by flash column chromatography, wherein the colorless oil is compound 2; S3. Compound 2 is dissolved in anhydrous ethanol, a phenylhydrazine derivative and p-toluenesulfonic acid are added, and the mixture is refluxed to obtain a second reaction solution; S4. After cooling the second reaction solution, saturated sodium bicarbonate was added to quench the reaction, extracted with ethyl acetate, 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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