阅读说明：本技术 双吡啶酮腙-n-甲基-3-吲哚甲醛西弗碱的用途 (Application of bispyridone hydrazone-N-methyl-3-indole formaldehyde Schiff base ) 是由 谭学杰 秦玥 于志杰 李维栋 孔曼 邢殿香 于 2021-11-26 设计创作，主要内容包括：双吡啶酮腙-N-甲基-3-吲哚甲醛西弗碱的用途本发明涉及药物化学、功能材料、荧光探针和废水处理领域,具体为双吡啶酮腙与N-甲基-3-吲哚甲醛反应生成的西弗碱在抗癌药物、抗菌药物、医药材料、功能材料及污水处理中的用途。双吡啶酮腙-N-甲基-3-吲哚甲醛西弗碱外观呈橙黄色棒状晶体,熔点为170-172℃,分子式为C-(21)H-(17)N-(5),结构如下：。该化合物对小鼠乳腺癌细胞4T-(1)的IC50为8.9μM；对人肝癌Hep G2细胞的IC50为7.5μM,表明其具有较好的肿瘤抑制活性；另外,该化合物也可以有效抑制大肠杆菌和金黄色葡萄球菌的繁殖。另外,该化合物与镉离子(Cd～(2+))可以生成具有强烈荧光的配合物沉淀,沉淀发出的荧光肉眼明显可见,该化合物可用于检测Cd～(2+)及去除废水中的镉离子。(The invention relates to the fields of medicinal chemistry, functional materials, fluorescent probes and wastewater treatment, in particular to application of Schiff base generated by reaction of bispyridone hydrazone and N-methyl-3-indole carbaldehyde in anticancer drugs, antibacterial drugs, medical materials, functional materials and sewage treatment. Bispyridone hydrazone-The appearance of N-methyl-3-indoxyl Schiff base is orange rod-shaped crystal, the melting point is 170-172 ℃, and the molecular formula is C 21 H 17 N 5 The structure is as follows: . The compound can be used for treating mouse breast cancer cell 4T 1 IC50 of (1) was 8.9. mu.M; IC50 of human liver cancer Hep G2 cells is 7.5 mu M, which shows that the human liver cancer Hep G2 cells have better tumor inhibition activity; in addition, the compound can also effectively inhibit the propagation of escherichia coli and staphylococcus aureus. In addition, the compound is mixed with cadmium ion (Cd) 2+ ) Can generate complex precipitate with strong fluorescence, the fluorescence emitted by the precipitate is obvious to the naked eye, and the compound can be used for detecting Cd 2+ And removing cadmium ions in the wastewater.)

1. The application of the compound shown as the following formula or the pharmaceutically acceptable salt thereof in preparing the medicine for preventing and/or treating tumors is characterized in that: the tumor is breast cancer or liver cancer; the appearance of the bipyridone hydrazone-N-methyl-3-indole formaldehyde Schiff base is orange rod-shaped crystal,the melting point is 170-172 ℃, and the molecular formula is C₂₁H₁₇N₅The structure is as follows:

。

2. the use of a compound as claimed in claim 1 in an antibacterial medicament or material, which is effective in inhibiting the proliferation of escherichia coli and staphylococcus aureus.

3. Use of a compound as claimed in claim 1 in a metal ion probe and in the treatment of waste water containing metal ions, wherein the metal ions are cadmium ions.

Technical Field

The invention relates to the fields of medicinal chemistry, functional materials, fluorescent probes and wastewater treatment, in particular to application of Schiff base generated by reaction of bispyridone hydrazone and N-methyl-3-indole carbaldehyde in anti-cancer drugs, antibacterial drugs, medical materials, functional materials, metal ion detection and wastewater treatment.

Background

Schiff base is a compound containing an imine or azomethine group, of the general formula R¹R²C=NR³. The Schiff base ligand with various structures and functions can be synthesized by introducing different substituent groups through the reaction design of a compound of an active carbonyl group and different amine compounds. Schiff base has many well-known obvious biological activities, such as antibiosis, antifungal, weeding, antituberculosis, anti-HIV, anticancer and the like, which makes it widely used in the fields of medicinal chemistry, functional materials and the like. In addition, Schiff bases having a large conjugated system tend to have very good fluorescent properties and are used in organic pigments, optical brighteners, photo-oxidizers, coatingsChemical and biochemical analysis, solar energy trap, anti-fake mark, medicine tracing, laser and other fields. In addition, the structure of the Schiff base is variable, the Schiff base can be coordinated with specific metal ions through regulating and controlling the structure, and the application of the Schiff base in multiple fields can be expanded according to the property of the complex, such as the fields of metal ion detection, sewage treatment, heavy metal detoxification drugs and the like. Therefore, the synthesis and property research of the novel Schiff base have great significance.

The invention content is as follows:

the invention relates to a synthetic method of Schiff base, namely Schiff base generated by reaction of bispyridone hydrazone and N-methyl-3-indole carbaldehyde, which is systematically named as follows: 3- ((di (pyridin-2-yl) methylene) hydrazone) methyl) -1-methyl-1H-indole with the molecular formula C₂₁H₁₇N₅(ii) a The appearance of the Schiff base is orange rod-shaped crystals, and the melting point is 170-172 ℃; the structure is as follows.

。

And identifying the structure.

Elemental analysis of bispyridone hydrazone-N-methyl-3-indolecarboxaldehyde Schiff base showed that C, H, N% of it is 74.35%, 5.09% and 20.66% respectively (theoretical values are 74.32%, 5.05% and 20.63% respectively), and the molecular weight is 339.402; the analysis of the single crystal structure shows that the crystal is monoclinic,P2₁/nspatial group, a =7.3653(5) a, b = 13.3069(11) a, c = 17.6980(11) a, α = γ =90 °, β =90 °, V =1734.6(2) a³Z =4, related¹HNMR spectrum,¹³The CNMR spectra are shown in figure 1 and figure 2 respectively; the structure diagram and the stacking diagram of the single crystal structure are shown in the attached figures 3 and 4 respectively.

And a synthetic method.

The bipyridyl ketone hydrazone-N-methyl-3-indole formaldehyde Schiff base has two synthesis methods: the first method is to take bipyridone hydrazone and N-methyl-3-indole formaldehyde as raw materials and comprises the following steps.

1) Dissolving the dipyridone hydrazone in a proper organic solvent, adding the N-methyl-3-indole formaldehyde according to a certain substance amount ratio, and stirring and reacting for a certain time at a certain temperature to complete the reaction.

The first step can also adopt solid phase reaction, namely organic solvent is not used, the dipyridone hydrazone and the N-methyl-3-indole formaldehyde are mixed according to a certain substance quantity ratio and then are ground and reacted for a certain time at normal temperature, and the obtained solid is the crude product of the target product.

2) If liquid phase reaction is adopted, filtering is carried out after the reaction is finished, and after the filtrate is naturally volatilized, the separated orange rod-shaped crystal is the target product; if solid phase reaction is adopted, the target product can be obtained by recrystallization with proper organic solvent after the reaction is finished.

The second method takes the bipyridone and the N-methyl-3-indole formaldehyde hydrazone as raw materials, and is completed in one step in a proper organic solvent, and the steps are as follows.

1) Dissolving dipyridone in a proper organic solvent, adding N-methyl-3-indole formaldehyde hydrazone according to a certain substance amount ratio, and stirring and reacting at a certain temperature for a certain time to complete the reaction; if the solid-phase reaction is adopted, an organic solvent is not used, the two are mixed according to a certain proportion and then the grinding reaction is carried out for a certain time, and the obtained solid is the crude product of the target product.

2) If liquid phase reaction is adopted, filtering is carried out after the reaction is finished, and after the filtrate is naturally volatilized, the separated orange rod-shaped crystal is the target product; if solid phase reaction is adopted, the target product can be obtained by recrystallization with proper organic solvent after the reaction is finished.

The two preparation methods are different mainly in reactants, but the molar ratio of the reaction raw materials is between 4:1 and 1: 4.

The organic solvent (including the solvent for reaction or the solvent for recrystallization) in the above two preparation methods is selected from: methanol, ethanol, acetonitrile, dichloromethane, chloroform, tetrahydrofuran, acetone, ethyl acetate, toluene, dioxane, etc.; the solid phase reaction does not use organic solvent, the raw materials react directly, and the solvent used for recrystallization can also be selected from the above solvents.

Preferably, the reaction temperature of the liquid phase reaction is normal temperature or heating reflux, and the reaction method is stirring; the solid phase reaction can be carried out at normal temperature, and the reaction method is grinding.

Preferably, the reaction time is selected from: 0.5-10 h.

The invention has the beneficial effects that: can synthesize more complex functional molecular materials by simpler steps and reactants.

And in vitro antitumor activity.

Mice growing in log phase were breast cancer 4T₁The cells or human hepatoma Hep G2 cells were digested with 0.25% pancreatic enzyme to become single cells, and prepared into 1.25X 10 by using 10% fetal bovine serum-containing F12K culture medium⁷One cell/L suspension of single cells, cells were seeded in 96-well plates at 200. mu.L per well (2.5X 10 per well)³Individual cells). Place 96-well cell culture plates in CO₂In an incubator at 37 ℃ with 5% CO₂Culturing for 48h under the condition.

When the cells in the wells are full (90% full), adding different doses of Schiff's base solution (200. mu.L/well) according to experimental groups to make the final concentrations of the compounds to be tested respectively 5. mu.M, 10. mu.M, 30. mu.M, 50. mu.M and 100. mu.M, setting 3 multiple wells in each group, and culturing for 96 h.

mu.L of MTT at a concentration of 0.5g/L was added to each well, and the culture was continued for 4 hours to reduce MTT to Formazan (Formazan). After all the supernatants were aspirated, 200. mu.L of DMSO was added to each well, and the mixture was shaken for 15min to dissolve formazan sufficiently, and then absorbance (OD value) at 490nm was measured using an ELISA detector. Then, the calculation was performed as follows.

Cell inhibition% = (control OD value-experimental OD value)/control OD value × 100%.

Test results show that the dipyridone hydrazone-N-methyl-3-indole formaldehyde Schiff base is used for 4T of mouse breast cancer cells₁IC50 (half maximal inhibitory concentration of drug) of 8.9 μ M; IC50 of Hep G2 cell of human liver cancer is 7.5 μ M; the inhibition rate of human foreskin fibroblast HFF-1 is 90.0 mu M; this indicates that the Schiff base molecule has a very good inhibitory effect on both cancer cells, and an inhibitory effect (IC) of cisplatin on both cancer cells₅₀=3.5 μ M and 4.9 μ MM) are comparable, while the inhibition rate on normal cells is lower.

And antibacterial activity.

The antibacterial effect is measured by adopting a filter paper diffusion method: soaking circular filter paper sheets (each sheet absorbs 10 microliters of liquid medicine) with the same size and the same diameter of 8mm into DMSO (dimethylsulfoxide) solution of a to-be-detected medicine with the concentration of 100 mug/mL, taking out the filter paper sheets after 30 minutes, airing, wherein the drug-loading capacity of each filter paper sheet is about 1 mug, then placing the filter paper sheets into the center of a plate coated with escherichia coli or staphylococcus aureus, pasting a label on a culture dish cover, marking, placing the culture dish into a constant-temperature incubator, culturing for 24 hours at 37 ℃, measuring the diameter of an antibacterial ring by using a vernier caliper, and comparing with penicillin potassium (the concentration of 10 mug/mL).

Test results show that the diameters of inhibition zones of the bispyridone hydrazone-N-methyl-3-indole formaldehyde Schiff base on escherichia coli/staphylococcus aureus are 11.0 mm and 12.5mm respectively; although smaller than 29.0 and 28.4mm of penicillin potassium, still has certain bacteriostatic activity.

Fluorescent probes and wastewater treatment.

Preparing dipyridone hydrazone-N-methyl-3-indolylcarbaldehyde Schiff base into 0.01M ethanol solution, respectively dropping the ethanol solution into 0.01M aqueous solution of the following 21 metal cations, Ca²⁺、Mg²⁺、Zn²⁺、Al³⁺、Fe³⁺、Pb²⁺、Ba²⁺、Cu²⁺、Cr³⁺、Mn²⁺、Ag⁺、Co²⁺、Ni²⁺、K⁺、Na⁺、Sn⁴⁺、Hg²⁺、Cd²⁺、La³⁺、Sm³⁺、Gd³⁺Of only Zn²⁺、Pb²⁺、Cu²⁺、Mn²⁺、Ag⁺、Co²⁺、Ni²⁺、Sn⁴⁺、Hg²⁺、Cd²⁺Precipitating 10 ions, centrifuging, and observing under 365nm ultraviolet lamp to obtain Cd²⁺The precipitate formed can be observed with naked eyes as strong yellow-green fluorescence (see figure 5); this indicates that the bipyridone hydrazone-N-methyl-3-indole carbaldehyde Schiff base can be used as Cd²⁺The specific fluorescent probe of (1) for detecting Cd²⁺Is present.

Dripping 0.01M bispyridone hydrazone-N-methyl-3-indoleformaldehyde Schiff base ethanol solution into the mixed solution of the 21 metal ions to generate precipitate, performing centrifugal separation, observing under a 365nm ultraviolet lamp, and observing strong yellow green fluorescence with naked eyes if mixed metal ions lack Cd²⁺No fluorescence could be observed with the naked eye (see fig. 6); this indicates bispyridone hydrazone-N-methyl-3-indolecarboxaldehyde Schiff base as Cd²⁺The fluorescent probe has stronger anti-interference performance.

Further experiments show that 1mL of 0.1M bispyridone hydrazone-N-methyl-3-indole formaldehyde Schiff base ethanol solution is dropped into 10mL of 0.01M Cd²⁺In the aqueous solution, after the reaction is finished, the Cd in the solution is remained²⁺Is less than 10^-4M; this shows that the bipyridone hydrazone-N-methyl-3-indole formaldehyde Schiff base can be used as Cd²⁺For Cd in an aqueous solution²⁺The Cd in the aqueous solution can be directly identified and simultaneously²⁺The removal is carried out, thereby realizing the one-step method for identifying and removing Cd in the aqueous solution^{2 +}The purpose of (2) is to simplify the operation process.

Drawings

FIG. 1 shows the preparation of bispyridone hydrazone-N-methyl-3-indolecarboxaldehyde Schiff base¹H NMR spectrum (solvent DMSO-d 6).

FIG. 2 shows the preparation of bispyridone hydrazone-N-methyl-3-indolecarboxaldehyde Schiff base¹³CNMR spectrum (solvent DMSO-d 6).

FIG. 3 is a crystal structure diagram of benzildihydrazone-N, N' -bis (3-formylindole) with an ellipsoid probability of 30%.

FIG. 4 is a packing diagram of the crystal structure of benzildihydrazone-N, N' -bis (3-formylindole) with 10% probability of ellipsoids.

FIG. 5 shows the observation of 10 precipitated metal ion complexes in the presence of 21 metal ions in benzildihydrazone-N, N' -bis (3-formylindole) under 365nm UV light.

FIG. 6 shows the phenomenon that after benzildihydrazone-N, N' -bis (3-formylindole) was added to a metal ion mixed solution, the precipitate appeared under a 365nm ultraviolet lamp, the left picture contains Cd²⁺Right picture without Cd²⁺。

Detailed Description

In order to better understand the present invention, the following embodiment further illustrates the technical solution of the present invention.

Example 1.

Weighing 1.98g (0.01 mol) of bispyridone hydrazone powder, placing the powder in a 100mL single-neck flask, pouring 30mL of toluene into the flask, stirring and dissolving at 50 ℃, weighing 1.45g (0.01 mol) of N-methyl-3-indole formaldehyde after the toluene is completely dissolved, adding the N-methyl-3-indole formaldehyde into the flask, heating to 80 ℃, refluxing and stirring for reaction for 8 hours, cooling to room temperature, filtering, placing the filtrate in a beaker for natural volatilization, and obtaining orange-yellow rod-shaped crystals at the bottom of the beaker after about 1 week, namely the bispyridone hydrazone-N-methyl-3-indole formaldehyde Schiff base which can be recrystallized by methanol to obtain a purer product.

Example 2.

Dissolving 1.84g (0.01 mol) of bipyridone in 30ml of ethyl acetate, adding 1.59g of N-methyl-3-indolylcarbaldehyde hydrazone into the ethyl acetate according to the mass ratio of 1:1, heating and refluxing for 3h, cooling to room temperature, filtering, placing the filtrate in a beaker for natural volatilization, and obtaining orange-yellow rod-shaped crystals, namely bipyridone hydrazone-N-methyl-3-indolylcarbaldehyde Schiff base, wherein the orange-yellow rod-shaped crystals can be obtained after the solvent is volatilized to be about 5ml, and the pure product can be obtained by recrystallization with methanol.

Example 3.

1.84g (0.01 mol) of bipyridyl ketone and 1.59g (0.01 mol) of N-methyl-3-indole carbaldehyde hydrazone are mixed and ground for 8 hours to obtain an orange rod-shaped crystal, namely bipyridyl ketone hydrazone-N-methyl-3-indole carbaldehyde Schiff base, and ethyl acetate can be used for recrystallization to obtain a purer product.