A ferrocene-piperazine hybrid compound, its preparation method and application

Abstract

The present invention discloses a ferrocene-piperazine hybrid compound, a preparation method thereof and an application thereof, belonging to the fields of compound synthesis and pharmaceutical technology. The preparation method is as follows: first, phosphorus trichloride and 3-ferrocenylpropionic acid are reacted in an organic solvent to prepare 3-ferrocenylpropionyl chloride; then 3-ferrocenylpropionyl chloride is reacted with 3-(2-ethoxyphenyl)-2-(1-(piperazin-1-yl)ethyl)quinazolin-4(3 H )-one to prepare the ferrocene-piperazine hybrid compound V. The synthesis method of the ferrocene-piperazine hybrid compound provided by the technical solution of the present invention is simple, and the hybrid compound shows micromolar anti-cancer activity and is expected to be used for preparing anti-cancer drugs.

Description

Technical Field

[0001] The present invention relates to the preparation and application of hybrid compounds, and specifically relates to a ferrocene-piperazine hybrid compound, a preparation method thereof, and an application thereof. Background Art

[0002] In the treatment of cancer, the challenges faced by currently used drugs are drug toxicity, high cost, and drug resistance. Ferrocene compounds are potential compounds with unique anti-cancer activity and are useful in modern organometallic chemistry industry due to the wide applications of ferrocene in catalysis, materials science, medicinal chemistry, and diagnostic applications.

[0003] Currently, some researchers have designed ferrocene-containing hybrid compounds with various biological activities, and these compounds exhibit many biological activities such as anti-malaria, anti-cancer, anti-tumor, anti-fungal, and anti-leishmaniasis activities. Due to the good stability of ferrocene groups towards water and air, low toxicity, unfamiliar redox activity, similarity, etc. in terms of aromaticity and chemical versatility, the efficacy of ferrocene hybrid compounds can be improved by hybrid compounds connected with other compounds through selected linkers (Molecules, 2019, 24(19):3604).

[0004] On the other hand, piperazine, as an organic compound, consists of a six-membered ring and contains two nitrogen atoms at opposite positions of the ring. The piperazine ring is a widespread structural element in drug discovery and has achieved extensive success in biological applications such as drug design. In addition, as an effective and specific ligand for a series of different scientific targets in medicinal chemistry, the piperazine moiety is regarded as a preferred framework. The piperazine scaffold is considered as a core and often appears in the natural active substances of various different therapeutic drugs (Expert Opin Ther Pat, 2016, 26(7):777). Piperazine and its derivatives have attracted people's attention in medicinal chemistry and modern drug discovery because it exhibits various biological activities, including anti-cancer activity. Therefore, the hybridization of ferrocene and piperazine is a promising strategy for developing new anti-cancer candidate drugs. Summary of the Invention

[0005] The present invention provides a novel ferrocene-piperazine hybrid compound, a preparation method thereof, and an application thereof.

[0006] A ferrocene-piperazine hybrid compound of the present invention has a structure shown in the following formula V:

[0007]

[0008] The preparation method of the hybrid compound with the structure shown in the above formula V has the following preparation route:

[0009]

[0010] The preparation method comprises the following steps:

[0011] (a) Take the compound shown by formula I and the compound shown by formula II, place them in an organic solvent, and heat under reflux to obtain the compound shown by formula III;

[0012] (b) Take the ferrocene compound shown by formula III and the compound shown by formula IV, place them in an organic solvent containing triethylamine, and carry out an acylation reaction to obtain compound V.

[0013] In step (a) of the above preparation method, the molar ratio of the compound shown by formula I to the compound shown by formula II is usually 1:20 - 25;

[0014] In step (a), the organic solvent is one or a combination of two or more of benzene, toluene and pyridine. The amount of the organic solvent used should be appropriate to dissolve the raw materials participating in the reaction. Usually, based on 1 mmol of the compound shown by formula 1, all the raw materials participating in the reaction can usually be dissolved in 40 - 100 mL of the above organic solvent. The reaction temperature is preferably carried out under heating conditions, and the heating temperature is 58 - 62 °C for reflux. The reaction time is preferably controlled within 3 h - 5 h.

[0015] In step (b) of the above preparation method, the molar ratio of the compound shown by formula III to the compound shown by formula IV is 1:0.7 - 0.9;

[0016] The molar ratio of the compound shown by formula III to triethylamine is 1:1.1 - 1.5;

[0017] In step (b), the organic solvent is specifically one or a combined solution of two of chloroform and dichloromethane; the reaction temperature is room temperature; triethylamine is used as a catalyst. The test results show that when the organic solvent is dichloromethane, a higher yield can be obtained. The amount of the organic solvent used should be appropriate to dissolve the raw materials participating in the reaction.

[0018] In step (b) of the above preparation method, the preferred reaction temperature is carried out at room temperature, and TLC is used to monitor whether the reaction is complete during the reaction process.

[0019] In step (b) of the above preparation method, the raw material compound shown by formula IV involved can be prepared with reference to the existing literature (BRENTR. Erastin analogs and uses thereof: US20070161644A1[P]. 2007 or Zhao Yanli et al. Synthesis of ferroptosis activator Erastin, Chinese Journal of Synthetic Chemistry 27.3 (2019): 4.), or can be prepared by designing its own synthesis route.

[0020] The applicant used 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method to detect the activity of the hybrid compound with the structure shown in V against cancer cells. The results showed that the compound shown in V had good inhibitory activity against triple-negative breast cancer cell lines MDA-MB-231, MDA-MB-468 and lung cancer cell A549, and exhibited better anti-cancer activity than the similar compound Erastin. Therefore, the present invention also provides the application of the hybrid compound with the structure shown in the above formula V in the preparation of anti-cancer drugs.

[0021] Compared with the prior art, the method for synthesizing the ferrocene-piperazine hybrid compound provided by the present invention is simple. The test results of the applicant showed that the ferrocene-piperazine hybrid compound had good inhibitory activity against cancer cells, was expected to be used for treating tumors, and had good potential medicinal value. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 For the 1 1H NMR spectrum of Example compound V;

[0023] Figure 2 For the anti-tumor activity diagram (IC 50 value, 48 h) of Example compound V against MDA-MB-231, MDA-MB-468 and A549 cancer cells. DETAILED DESCRIPTION OF THE INVENTION

[0024] In order to better explain the technical solution of the present invention, the content of the present invention will be further described in detail below in conjunction with embodiments, but the embodiments of the present invention are not limited thereto.

[0025] Embodiment

[0026] The preparation and characterization of a ferrocene-piperazine hybrid compound, and the preparation route is as follows:

[0027]

[0028] (a) Benzene, 58 - 62 °C, 3 - 5 h;

[0029] (b) Organic solvents such as dichloromethane and chloroform, triethylamine, at room temperature.

[0030] The specific preparation method is as follows:

[0031] (a) Compound II (5 ml, 50 mmol) was added to a benzene solution of 3-ferrocenylpropionic acid (500 mg, 1.9 mmol) in 100 ml. The above solution was refluxed at 58 - 62 °C, and the reaction was monitored by TLC. The reaction mixture was stirred for 3 - 5 h; then, after filtering off the precipitate, the filtrate was evaporated to dryness under reduced pressure to obtain the red oily compound III; MS m / z: 276; elemental analysis: (%) C 13 H 13 ClFeO: calculated: C, 56.4; H, 4.7, found: C 56.1; H 4.9;

[0032] (b) Under normal temperature conditions, Compound III (1 mmol) and Compound IV (0.8 mmol) were dissolved in 5 ml of dichloromethane, and then triethylamine (1.2 mmol) was added. The reaction was stirred at room temperature for 2 h and washed with saturated aqueous sodium bicarbonate solution; then the crude product was purified by column chromatography to obtain the corresponding hybrid compound V.

[0033] Refer to Figure 1 , Compound V and its characterization are as follows:

[0034]

[0035] 1 H NMR (400 MHz, Chloroform-d) δ 8.22 (d, J = 7.9 Hz, 1H), 7.71 (d, J = 3.9 Hz, 2H), 7.46–7.32 (m, 2H), 7.07 (d, J = 7.4 Hz, 1H), 7.03–6.93 (m, 2H), 5.23 (s, 2H), 4.19 (s, 2H), 3.98 (d, J = 6.4 Hz, 2H), 3.42 (s, 1H), 3.32 (s, 1H), 3.14 (s, 1H), 2.74 (s, 2H), 2.46 (s, 2H), 2.37 (d, J = 23.5 Hz, 3H), 2.09 (s, 1H), 1.26 (d, J = 6.5 Hz, 3H), 1.18 (t, J = 13.3 Hz, 6H), -0.07 (s, 4H);

[0036] Elemental analysis: (%) C 35 H 38 FeN 4 O 3 : calculated: C, 67.9; H, 6.2; N, 9.0, found: C 67.1; H 6.4; N 8.6.

[0037] The applicant conducted an in vitro anti-cancer activity study on the prepared Compound V:

[0038] The in vitro anti-cancer activity of Compound V against triple-negative breast cancer cell lines MDA-MB-231, MDA-MB-468 and human lung cancer cell line A549 was determined by the MTT assay. The above cells were maintained in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS), 50 U / mL penicillin, and 50 mg / mL streptomycin, and incubated at 37 °C in an atmosphere of 5% carbon dioxide. Cells in the logarithmic growth phase with good cell status were selected for the cytotoxicity experiment. 100 μL of cell suspension (density of 5×10 4 cells / mL) was seeded in 96-well plates in quintuplicate and cultured at 37 °C in an atmosphere of 5% carbon dioxide for 24 hours. Then the medium was removed and replaced with the corresponding medium containing 10% FBS with an appropriate concentration of the compound, and cultured under the conditions of 37 °C and 5% CO 2 for 72 hours. 10 μL of MTT dye (5 mg / mL) was added to each well. After incubation for another 4 hours, the absorbance at 570 nm was read using an enzyme-linked immunosorbent assay reader. Cytotoxicity was evaluated based on the percentage of cell survival compared to the negative control. The final IC 50 value was calculated by the Bliss method (n = 5).

[0039] As can be seen from Figure 2 , the target compound V showed micromolar anti-cancer activity against MDA-MB-231, MDA-MB-468 and A549 cancer cells, and was superior to the anti-cancer activity of the cell death activator Erastin (structurally similar to the target compound V), indicating that this compound V has prospects for tumor treatment.

Claims

1. A ferrocene-piperazine hybrid compound, Characterized in that: The structure of the hybrid compound is shown in the following formula: 。 2. The preparation method of the ferrocene-piperazine hybrid compound according to claim 1, Characterized in that, The preparation route is as follows: ； The preparation method includes the following steps: (a) Take the compound shown in formula I and the compound shown in formula II and place them in an organic solvent, heat under reflux to obtain the compound shown in formula III; (b) Take the compound shown in formula III and the compound shown in formula IV, place them in an organic solvent containing triethylamine, and carry out an acylation reaction to obtain the hybrid compound V.

3. According to the preparation method described in claim 2, Characterized in that: In step (a), the molar ratio of the compound shown in formula I to the compound shown in formula II is 1:20 - 25; In step (a), the organic solvent is one or a combination of two or more of benzene, toluene and pyridine; The reaction temperature is heated under reflux at 58 - 62 °C, and the reaction time is 3 - 5 h.

4. According to the preparation method described in claim 3, Characterized in that: In step (b), the molar ratio of the compound shown in formula III to the compound shown in formula IV is 1:0.7 - 0.9; The molar ratio of the compound shown in formula III to triethylamine is 1:1.1 - 1.5; In step (b), the organic solvent is one or a combined solution of chloroform and dichloromethane; The reaction temperature is room temperature; triethylamine is used as a catalyst.

5. The application of the ferrocene-piperazine hybrid compound according to claim 1 in the preparation of drugs for treating triple-negative breast cancer and lung cancer.

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