CN106543208A - Copper chloride (II) chelate and its synthetic method and application with 1 pyridine β carbolines as part - Google Patents

Abstract

The invention discloses a copper (II) chloride chelate with 1-pyridine-β-carboline as a ligand, a synthesis method and an application thereof. The structural formula of this copper chloride (II) chelate compound is shown in the following formula (I), and its preparation method is: take the compound shown in the following formula (II) and copper chloride dihydrate, dissolve in a polar solvent, and prepare Bit reaction, that is. The copper (II) chloride chelate compound of the present invention exhibits stronger antitumor activity than ligands and cisplatin, has good potential medical value, and is expected to be used in the preparation of various antitumor drugs. The structures represented by formula (I) and formula (II) are as follows.

Description

Copper (II) chloride chelate with 1-pyridine-β-carboline as ligand and its synthesis method and application

technical field

The invention relates to the technical field of medicine, in particular to a copper (II) chloride chelate with 1-pyridine-β-carboline as a ligand and a synthesis method and application thereof.

Background technique

Cancer (also known as malignant tumor) is a systemic and systemic disease involving multiple genes and gradually developing. Statistics show that cancer has become the number one cause of death in quite a few countries and regions, and it is showing a diversified and continuously increasing incidence trend. It is estimated that by 2020, the number of cancer patients worldwide will exceed 20 million, and this trend is developing China is even more serious. How to overcome cancer has become the goal of the medical community. With the development of medical science, people's understanding of tumors is deepening, and every link in the occurrence and development of tumors may become a potential target for treatment.

In recent years, with the clinical application of drugs such as cisplatin, metal-based chelate anticancer drugs have gradually become a research hotspot. The combination of the inherent properties of metals and bioactive ligand molecules provides a broad space for the development of new drugs with high efficiency, low toxicity, broad spectrum and targeted activity.

β-carboline alkaloids are a large class of alkaloids from natural sources, widely distributed in nature. Among them, the most common β-carboline alkaloids extracted from the seeds of the perennial herb of Tribulus terrestris, mainly include dehydrohaeline and haemaline. Relevant studies at home and abroad have found that β-carboline alkaloids exhibit broad-spectrum anti-tumor activity, and their toxic and side effects are relatively small. The structural modification of β-carboline alkaloids has always attracted the attention of researchers. It is generally believed that the planar conjugated structure of most β-carboline alkaloids and the existence of different substituent groups (such as benzyl (-C ₆ H ₅ ), methoxyl (-OCH ₃ )) have important effects on their antitumor activity. Significant relationship. The current research on β-carboline alkaloids mainly includes the following aspects: one is to find and purify from plants of different families and genera based on the chemical research of natural products; the other is to synthesize β-carbolines through organic synthesis. Alkaloids are fully or semi-synthesized to modify their structures. The other is to study the molecular mechanism of its pharmacological activity (such as anti-tumor activity). Such as inserting DNA, inhibiting topoisomerase, inhibiting CDK, etc. However, judging from the current research progress, the content of β-carboline alkaloids in natural plants is generally low, and the extraction is relatively complicated, which is not conducive to in-depth research on it. Although it has certain advantages, it is limited by the cost of preparation, so the current research on the expansion of β-carboline alkaloids is still insufficient.

On the other hand, although cisplatin has been successfully marketed for decades and successfully treated a variety of cancers, clinical results show that it still has some problems, such as drug resistance and toxic side effects, and some cancers naturally show resistance to cisplatin , and some cancers gradually show induced drug resistance after initial treatment. Both preclinical and clinical experiments have shown that non-platinum antitumor drugs have broad prospects for development, but the current research on copper(II) chloride chelates with 1-pyridine-β-carboline as a ligand is still blank.

Contents of the invention

The technical problem to be solved by the present invention is to provide a novel structure copper (II) chloride chelate compound with 1-pyridine-β-carboline as a ligand, as well as its synthesis method and application.

The present invention relates to a compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof:

The synthesis method of the compound shown in the above formula (I) is: take the compound shown in the following formula (II) and copper chloride dihydrate (CuCl ₂ ·2H ₂ O), dissolve in a polar solvent, and carry out a coordination reaction, namely get the target product;

The synthetic route of above-mentioned synthetic method is as follows:

The raw material formula (II) involved in the above synthesis method participates in the reaction as a ligand, and its chemical name is 1-pyridine-β-carboline (1-Pyridine-β-Carboline), referred to as KL. The compound can be prepared by designing a synthetic route by itself, preferably by the following method: using tryptamine and pyridine-2-carbaldehyde as raw materials, reacting in the first organic solvent, and obtaining compound 1 through dehydration condensation; then placing compound 1 in In the second organic solvent, add an oxidizing agent to close the ring and dehydrogenate to obtain; wherein:

The first organic solvent is one or a combination of two or more selected from toluene, methanol, ethanol, methylene chloride and chloroform;

The second organic solvent is one or a combination of two or more selected from benzene, toluene, p-xylene, glacial acetic acid and methylene chloride;

The oxidizing agent is palladium carbon, manganese acetate hydrate (Mn(Ac) ₃ ·nH ₂ O), lead tetraacetate (Pb(Ac) ₄ ) or 2,3-dichloro-5,6-dicyano-p-phenylene quinone.

The synthetic route of compound method shown in above-mentioned preparation formula (II) is as follows:

Reagents: (a) first organic solvent; (b) oxidizing agent, second organic solvent.

A more specific synthetic method of the compound shown in the above formula (II), comprising the following steps:

① Use tryptamine and pyridine-2-carbaldehyde as raw materials, react in the first organic solvent, discharge the water generated by the reaction during the reaction, evaporate the solvent to dryness after the reaction, and obtain compound 1;

② Place compound 1 in a second organic solvent, add an oxidizing agent, and react under heating conditions. After the reaction is completed, filter, collect the filtrate, and evaporate to dryness to obtain compound 2 represented by formula (II).

In step ① of the compound synthesis method shown in the above formula (II), the ratio of the amount of tryptamine to pyridine-2-carboxaldehyde is usually 0.8 to 1.2:1, and the reaction can be carried out under heating or without heating. During the process, a water separator can be used to discharge the water generated by the reaction, and whether the reaction is complete can be tracked and detected by thin layer chromatography (TLC); preferably, the reaction is heated and refluxed, and the reaction time is controlled at 2 to 6 hours. . In this step, the crude product of compound 1 is obtained. In order to reduce the impurities in the subsequent reaction and increase the yield of the subsequent reaction, it is preferable to carry out the subsequent reaction after purifying the obtained residue. The specific purification operation may be to recrystallize the obtained residue with a small polar solvent, and the obtained recrystallized product is used for subsequent reactions. The low-polarity solvent used for recrystallization is the same as the prior art, specifically petroleum ether and/or n-hexane and the like.

In the step ② of the synthesis method of the compound represented by the above formula (II), the reaction is preferably heated to reflux, and whether the reaction is complete can be detected by thin-layer chromatography. In this step, according to the difference of oxidizing agent, select different second organic solvents for use, specifically as follows:

(1) When the selection of the oxidizing agent is palladium carbon, the second organic solvent is preferably one or more combinations of benzene, toluene and p-xylene, and when the selection of the second organic solvent is the combination of the above two or more , the ratio between them can be any ratio. The amount of palladium carbon added is usually 2-3 g of palladium carbon according to 10 mmol of compound 1, and the palladium carbon can be 5% Pd/C or 10% Pd/C.

(2) When the selection of the oxidizing agent was manganese acetate hydrate or lead tetraacetate, the second organic solvent was preferably glacial acetic acid; the addition of the manganese acetate hydrate or lead tetraacetate was usually 2-2% of the amount of compound 1 substance. 8 times. When the oxidizing agent is manganese acetate hydrate or lead tetraacetate, it is preferable to adjust the pH of the system with lye to ≥7 after the reaction, and then extract it, collect the organic phase, evaporate the residue obtained from the solvent, and then apply the silica gel column layer analysis and purification; wherein, the lye can be an aqueous solution of alkaline substances such as ammonia, sodium acetate, sodium carbonate, sodium phosphate, sodium bicarbonate or potassium carbonate, and the concentration of the lye is preferably 5 to 30/w/ w%; the solvent used to extract the system after adjusting the pH value can specifically be ethyl acetate, dichloromethane, chloroform or ether.

(3) When the selection of the oxidant is 2,3-dichloro-5,6-dicyano-p-benzoquinone, the second organic solvent is preferably one or more than two combinations of benzene, toluene and methylene chloride, when When the selection of the two organic solvents is a combination of the above two or more, the ratio between them can be any ratio. The amount of 2,3-dichloro-5,6-dicyano-p-benzoquinone added is usually 1 to 4 times the amount of compound 1.

What the above-mentioned method prepares is the crude product of the compound shown in formula (II), in order to further improve its purity, be conducive to the carrying out of follow-up reaction, preferably above-mentioned gained crude product is purified, purified and then used for the synthesis of the target product of the present invention method. The described purification operation is the same as the prior art, and the specific purification operation can be to recrystallize the obtained residue with methanol, or to purify the obtained residue by silica gel column chromatography. When the silica gel column chromatography is used, the eluent It is a mixed solvent composed of petroleum ether and methylene chloride at a volume ratio of 6:1 to 1:1.

In the synthetic method of the compound represented by the formula (I) of the present invention, the polar solvent is methanol and selected from water, acetone, chloroform, dimethylsulfoxide, N,N-dimethylformamide and acetonitrile one or any combination of two or more. Preferably, the proportion of methanol in the polar solvent is 70-85v/v%. When the polar solvent contains any two or more of water, acetone, chloroform, dimethyl sulfoxide, N,N-dimethylformamide and acetonitrile, on the premise that their total amount does not exceed 30% Under certain conditions, their ratio can be any ratio. The amount of the polar solvent can be determined according to the needs. Usually, 1 mmol of the compound represented by formula (II) and 1 mmol of copper chloride dihydrate are dissolved in 35-40 mL of polar solvent. In the specific dissolving step, generally the compound shown in formula (II) and cupric chloride dihydrate are mixed and then a polar solvent is added; the compound shown in formula (II) and cupric chloride dihydrate can also be respectively The solvent is dissolved, and then mixed for reaction.

In the synthesis method of the compound represented by the formula (I) of the present invention, the molar ratio of the copper chloride dihydrate to the compound represented by the formula (II) is a stoichiometric ratio of 1:1.

The compound represented by the formula (I) of the present invention can be synthesized by normal pressure solution method or high pressure solvothermal method when synthesizing.

When adopting normal pressure solution method, its synthetic method comprises: take compound shown in formula (II) and cupric chloride dihydrate to be dissolved in polar solvent, gained mixed solution is reacted under heating condition, reactant removes part solvent, static Place, precipitate, and separate the solid to obtain the target product.

In the above normal pressure solution method, the reaction preferably adopts reflux reaction, and the reaction is preferably carried out in the range of 45°C to the reflux temperature of the polar solvent, more preferably at 60-80°C. To judge whether the reaction is complete or not, thin-layer chromatography can be used for follow-up detection. Under the above limited conditions, it is more appropriate to control the reaction time within 24-48 hours. The reactant is concentrated to remove part of the solvent, usually 70-90% of the added amount of the polar solvent is removed by concentration.

When the high-pressure solvothermal method is adopted, the synthesis method comprises: dissolving the compound shown in formula (II) and cupric chloride dihydrate in a polar solvent, placing the resulting mixture in a container, freezing it with liquid nitrogen, and then pumping it to a vacuum , melted and sealed, and then reacted under heating conditions to obtain the target product.

In the above-mentioned high-pressure solvothermal method, the container is usually a thick-walled borosilicate glass tube, and the reaction temperature is usually carried out at 60-80°C. Under this temperature condition, the reaction time is preferably controlled at 48-72h. The actual situation extended to more than 72h.

The present invention also includes the use of the compound represented by the above formula (I) or a pharmaceutically acceptable salt thereof in the preparation of antitumor drugs.

The present invention also includes the antitumor drug prepared from the compound represented by the above formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient.

Compared with prior art, the present invention provides a kind of new cupric chloride (II) chelate, promptly take 1-pyridine-beta-carboline as the cupric chloride (II) chelate of ligand, and Its synthesis method and application. The applicant investigated its inhibitory effect on a variety of tumor cell lines, and the results showed that the chelate showed stronger antitumor activity than its ligand and cisplatin, and had good potential medicinal value, which is expected to be applied to various Preparation of an antitumor drug.

Description of drawings

Fig. 1 is the proton nuclear magnetic resonance spectrogram of the final product that the embodiment of the present invention 1 makes;

Fig. 2 is the carbon nuclear magnetic resonance spectrogram of the final product that the embodiment of the present invention 1 makes;

Fig. 3 is the electrospray mass spectrogram of the final product that the embodiment of the present invention 1 makes;

Fig. 4 is the X-ray single crystal structure figure of the final product that the embodiment 1 of the present invention makes;

Fig. 5 is the X-ray single crystal structure figure of the final product that the embodiment 5 of the present invention makes;

Fig. 6 is the electrospray mass spectrogram of the final product prepared in Example 5 of the present invention.

detailed description

The present invention will be described in further detail below in conjunction with specific examples to better understand the content of the present invention, but the present invention is not limited to the following examples.

Embodiment 1: the compound shown in formula (II) is the synthesis of 1-pyridine-beta-carboline (KL)

1) Dissolve 1.60g (10mmol) of tryptamine in 70mL of dichloromethane, then add 1.07g (10mmol) of pyridine-2-carbaldehyde, and heat to reflux for 8 hours; after the reaction, distill under reduced pressure to obtain the crude compound 1 ;

2) Dissolve 2.49g (10mmol) of compound 1 in 80mL of glacial acetic acid, then add 13.4g of manganese acetate hydrate (Mn(Ac) ₃ nH ₂ O), heat to 80°C, react overnight, evaporate the solvent to dryness, add 100mL of water, extracted 3 times with ethyl acetate, combined the organic phases, and evaporated the solvent to obtain an oily crude product, which was then purified by flash liquid chromatography (V _{petroleum ether} : V _{dichloromethane} = 1:1) to obtain light yellow crystals Compound 2 (~73% yield).

The resulting pale yellow crystals were subjected to proton nuclear magnetic resonance spectrum, carbon nuclear magnetic resonance spectrum, electrospray mass spectrometry and single crystal diffraction analysis. The specific spectral characteristics are as follows:

(1) Proton nuclear magnetic resonance spectrum and carbon spectrum, and their spectrograms are shown in Figures 1 and 2 respectively.

¹ H NMR (500MHz, CDCl ₃ ) δ: 11.22(s, 1H), 8.82～8.74(m, 2H), 8.53(d, J=5.1Hz, 1H), 7.98(d, J=5.1Hz, 1H) ,7.90(td,J=7.8,1.8Hz,1H),7.60(d,J=2.4Hz,1H),7.54(d,J=8.8Hz,1H),7.37–7.32(m,1H),7.25( dd, J=8.8, 2.5Hz, 1H), 3.96 (s, J=4.0Hz, 3H). 1H NMR (500MHz, DMSO-d6) δ11.97(s, 1H), 8.90(d, J=4.7Hz, 1H), 8.65(d, J=8.0Hz, 1H), 8.51(d, J=5.0Hz, 1H), 8.30(d, J=7.8Hz, 1H), 8.25(d, J=5.0Hz, 1H), 8.05(t, J=7.7Hz, 1H), 7.90(d, J=8.2Hz, 1H) , 7.60 (t, J = 7.7Hz, 1H), 7.53 (m, 1H), 7.29 (t, J = 7.5Hz, 1H).

¹³ C NMR(126MHz,CDCl ₃ )δ:157.87,154.08,148.25,138.09,137.44,136.83,135.68,135.30,130.36,122.92,121.41,121.35,118.51,115.36,112.67,103.57,77.30,77.25,77.04,76.79 , 56.02 13C NMR (150MHz, DMSO-d6) δ157.10, 148.72, 141.01, 138.08, 137.70, 137.24, 133.54, 129.90, 128.36, 123.38, 121.62, 120.83, 120.33, 119.591, 115

(2) Electrospray mass spectrometry, as shown in Figure 3, ESI-MS m/z: 246.10[M+H] ⁺ .

(3) X-ray single crystal diffraction analysis, to determine its single crystal structure as shown in Figure 4.

Therefore, it can be determined that the above-mentioned light yellow solid product is 1-pyridine-β-carboline, and its chemical structural formula is as shown in the following formula (II):

Embodiment 2: the synthesis of ligand KL

1) Dissolve 1.60 g (10 mmol) of tryptamine in 70 mL of toluene, then add 1.07 g (10 mmol) of pyridine-2-carbaldehyde, and heat to reflux for 6 hours. After the reaction, distill under reduced pressure to obtain the crude compound 1;

2) Dissolve 2.49g (10mmol) of compound 1 in 80mL of glacial acetic acid, then add 13.4g of manganese acetate hydrate (Mn(Ac) ₃ ·nH ₂ O), heat to 70°C, react overnight, evaporate the solvent to dryness, add 100mL of water, extracted 3 times with ethyl acetate, combined the organic phases, and evaporated the solvent to obtain an oily crude product, which was then purified by flash liquid chromatography (V _{petroleum ether} : V _{dichloromethane} = 2:3) to obtain light yellow crystals Compound 2 (~55% yield).

The obtained pale yellow crystals were analyzed by H NMR, C NMR, electrospray mass spectrometry and single crystal diffraction, and it was determined to be the target product 1-pyridine-β-carboline.

Embodiment 3: the synthesis of ligand KL

1) 1.60g (10mmol) tryptamine is dissolved in the mixed solvent that is made up of the methanol of 30mL and the ethanol of 40mL, then add 1.07g (10mmol) pyridine-2-carboxaldehyde, heat to reflux for 12 hours, after the reaction finishes, reduce Press distillation to obtain crude product compound 1;

2) Dissolve 2.49g (10mmol) of compound 1 in 80mL of glacial acetic acid, then add 20mmol of Pb(Ac) ₄ , heat to 90°C, react overnight, evaporate the solvent to dryness, add 100mL of water, and extract 3 times with ethyl acetate, The organic phases were combined, and the solvent was evaporated to dryness to obtain an oily crude product, which was then purified by flash liquid chromatography (V _{petroleum ether} : V _{dichloromethane} =3:2) to obtain light yellow crystal compound 2 (the yield was about 67%) .

The obtained pale yellow crystals were analyzed by H NMR, C NMR, electrospray mass spectrometry and single crystal diffraction, and it was determined to be the target product 1-pyridine-β-carboline.

Embodiment 4: the synthesis of ligand KL

1) Add 1.6g (10mmol) tryptamine, 1.1g (10mmol) pyridine-2-carboxaldehyde and 50ml chloroform to a 150ml round bottom flask, heat to reflux for 4 hours, evaporate the solvent after the reaction is over, and use 100ml Recrystallization from n-hexane gave compound 1 (2.3 g, yield 92%);

2) Add 2.5g (10mmol) compound 1, 2.5g palladium carbon (10%Pd/C) and 100ml p-xylene to a 250ml round bottom flask, heat to reflux, and use thin layer chromatography to track and detect until the end of the reaction, and let stand Suction filtration and the filtrate was evaporated to dryness, and the obtained residue was purified by silica gel column (chromatographic purification (V petroleum ether: V dichloromethane = 1:1) to obtain light yellow crystal compound 2 (2.1 g, yield 86%).

The obtained pale yellow crystals were analyzed by H NMR, C NMR, electrospray mass spectrometry and single crystal diffraction, and it was determined to be the target product 1-pyridine-β-carboline.

Embodiment 5: the synthesis of chelate [Cu ^II (KL) Cl ₂ ]

In a thick-walled borosilicate glass tube with an open end, directly add 0.1 mmol of copper chloride dihydrate and 0.1 mmol of ligand KL, and then add 1.5 mL of a mixed solvent consisting of methanol and dimethyl sulfoxide (the volume of methanol and acetonitrile The ratio is 4:1); after freezing with liquid nitrogen, pump it to a vacuum, seal the open end, and then fully react at 80°C for 72h; A green rod-like crystalline solid product was precipitated.

The above-mentioned yellow-green product obtained by electrospray mass spectrometry (as shown in Figure 6) and X-ray single crystal diffraction analysis structure (as shown in Figure 5) was determined to be a chlorine compound with 1-pyridine-β-carboline as a ligand. Copper (II) chelate, i.e. the target chelate [Cu ^II (KL) Cl ₂ ], its structural formula is as shown in the following formula (I):

Embodiment 6: the synthesis of chelate [Cu ^II (KL) Cl ₂ ]

Repeat embodiment 5, difference is:

The polar solvent was changed to a mixed solvent composed of methanol and water (the volume ratio of methanol and water was 8:2), the reaction temperature was changed to 60°C, and the reaction time was 72h. After the reaction was completed, it was directly cooled (yield 76%) .

The precipitated yellow-green product was determined to be the target chelate [Cu ^II (KL)Cl ₂ ] by electrospray mass spectrometry and X-ray single crystal diffraction analysis for structure determination.

Embodiment 7: the synthesis of chelate [Cu ^II (KL) Cl ₂ ]

Repeat embodiment 5, difference is:

The polar solvent was changed to a mixed solvent composed of methanol and chloroform (the volume ratio of methanol and chloroform was 5:1), the reaction temperature was still 80°C, and the reaction time was 72h. After the reaction was completed, it was directly cooled (68% yield) .

The precipitated yellow-green product was determined to be the target chelate [Cu ^II (KL)Cl ₂ ] by electrospray mass spectrometry and X-ray single crystal diffraction analysis for structure determination.

Embodiment 8: the synthesis of chelate [Cu ^II (KL) Cl ₂ ]

Weigh 0.1mmol of ligand KL and 0.1mmol of copper chloride dihydrate, dissolve ligand KL in 25mL of methanol, dissolve copper chloride dihydrate in 10mL of N,N-dimethylformamide, and mix well Two solutions; reacted at 80°C for 24h, distilled off most of the solvent (80% of the added amount of solvent) under reduced pressure, cooled to room temperature, stood still, precipitated a green solid, separated the solid, washed with water, and dried to obtain a green solid Product (79% yield).

The precipitated yellow-green product was determined to be the target chelate [Cu ^II (KL)Cl ₂ ] through electrospray mass spectrometry combined with X-ray single crystal diffraction analysis for structure determination.

Embodiment 9: the synthesis of chelate [Cu ^II (KL) Cl ₂ ]

Repeat embodiment 8, difference is:

N,N-dimethylformamide in the polar solvent was changed to acetone, wherein the volume ratio of methanol to acetone was 10:1, the reaction temperature was 75° C., and the reaction time was 48 h (yield 71%).

The precipitated yellow-green product was determined to be the target chelate [Cu ^II (KL)Cl ₂ ] through electrospray mass spectrometry combined with X-ray single crystal diffraction analysis for structure determination.

Embodiment 10: the synthesis of chelate [Cu ^II (KL) Cl ₂ ]

Weigh 0.1mmol of ligand KL and 0.1mmol of copper chloride dihydrate, dissolve ligand KL and copper chloride dihydrate in a mixed solvent consisting of 10mL methanol, 1mL acetonitrile and 1mL water, and react at 70°C for 48h , Most of the solvent was distilled off under reduced pressure (90% of the added amount of solvent), cooled to room temperature, left standing, and a green solid was precipitated. The solid was isolated, washed with water and dried to give the product as a green solid (75% yield).

The precipitated yellow-green product was determined to be the target chelate [Cu ^II (KL)Cl ₂ ] through electrospray mass spectrometry combined with X-ray single crystal diffraction analysis for structure determination.

In order to fully illustrate the use of copper chloride (II) chelates of the present invention as ligands in pharmacy with 1-pyridine-β-carboline, the applicant has carried out this chelate and its ligand KL Antitumor activity assay.

Experimental example 1: take 1-pyridine-β-carboline as the copper chloride (II) chelate compound (made by the method described in Example 5) and ligand KL (made by the method described in Example 1) of ligand In vitro inhibitory activity experiments on a variety of human tumor strains

1. Cell lines and cell culture

NCI-H-460 (human large cell lung cancer), SK-OV-3 (human ovarian adenocarcinoma cell line), Hep G2 (human liver cancer cell line), MGC803 (human gastric cancer cell line), T24 (human bladder cancer cell line) were used in this experiment. Wait.

All cell lines were cultured in DMEM medium containing 10% FBS fetal bovine serum and 1% penicillin and streptomycin mixture, and cultured in an incubator containing 5% CO ₂ at a volume concentration of 37°C.

2. Primary screening

After the 96-well plate of each cell strain adhered to the wall, it was replaced with serum-free DMEM, and the drug was added 12 to 18 hours later. Compounds used in this experiment (both purity ≥ 95%) were formulated to an intermediate concentration of 200 μmol/L, and 20 μL of the intermediate concentration compound solution was added to a 96-well plate with 180 μL of cell fluid, and the final concentration of the compound was 20 μmol/L. The final concentration of DMSO is ≤1%, and the inhibitory degree of the compound on tumor cell growth is tested at this concentration. If the inhibition rate is greater than 50%, and conforms to the morphological changes (such as cell shrinkage, fragmentation, floating, etc.) of the cells under the light microscope, it is judged that the primary screening is effective, and the results of the inhibition rate are as follows Table 1 shows.

3. Cell growth inhibition test (MTT method)

After dissolving the test compound to be tested in DMSO, dilute the intermediate concentration in serum-free medium to 200 μmol/L, 100 μmol/L, 50 μmol/L, 25 μmol/L, 12.5 μmol/L, and then use a microtube with a diameter of d=0.22 μm Sterilize by filtering through a pore filter and store at 4°C.

A series of tumor cell lines in the logarithmic growth phase were respectively inoculated in a 96-well plate at 180 μL per well, and 200 μL of PBS (phosphate-buffered saline powder preparation solution) was added to the edge wells. The cell concentration was about 1×10 ⁴ /well, and cultured for 12 After the cells adhered to the wall, change serum-free DMEM, 12 to 18 hours compound medium concentration liquid, set 5 duplicate wells in parallel for each concentration of the compound, wherein the final concentration of DMSO ≤ 1%, set the corresponding negative control group (cultivation There were only cells and the same amount of DMSO in the medium, no drug) and a blank control group (only the same amount of drug in the culture medium, no cells), and each group was also set up in parallel with 5 replicate wells, and the drug action time was 48h. Add 10 μL of MTT (5 mg/mL PBS) to each well 4 hours before the end of the culture, continue to culture for 4 hours, pour out the culture solution, add 100 μL/well of DMSO, and vibrate with a plate shaker for 7 minutes to fully dissolve the crystals, and set the blank control group to zero , using a microplate reader to measure the absorbance (A) value after removing the background light absorption value with a dual wavelength of 570nm/630nm, and measure the inhibition rate, IC ₅₀ , the test results are shown in Table 2 below:

Table 1:

Table 2:

Claims (10)

1. compound or its pharmaceutically acceptable salt shown in following formula (I): 2. the synthetic method of the described compound of claim 1 is characterized in that: get compound shown in following formula (II) and cupric chloride dihydrate, be dissolved in polar solvent, carry out coordination reaction, promptly obtain target product; 3. synthetic method according to claim 2, is characterized in that: described polar solvent is methyl alcohol and is selected from water, acetone, chloroform, dimethylsulfoxide, N, N-dimethylformamide and acetonitrile one or any combination of two or more. 4. according to the described synthetic method of claim 2 or 3, it is characterized in that: get compound shown in formula (II) and copper chloride dihydrate, be dissolved in polar solvent, gained mixed solution reacts under heating condition, reacts Part of the solvent was removed from the product, left to stand, precipitated, and the solid was separated to obtain the target product. 5. The synthetic method according to claim 4, characterized in that: the reaction is carried out at 60° C. to the reflux temperature range of the polar solvent. 6. according to the described synthetic method of claim 2 or 3, it is characterized in that: get compound shown in formula (II) and copper chloride dihydrate, be dissolved in polar solvent, gained mixed solution is placed in container, through liquid After being frozen with nitrogen, it was pumped to a vacuum, melted and sealed, and then reacted under heating conditions to obtain the target product. 7. The synthesis method according to claim 6, characterized in that: the reaction is carried out at 60-80°C. 8. according to the described synthetic method of claim 2 or 3, it is characterized in that: the compound shown in described formula (II) is prepared by the following method: take tryptamine and pyridine-2-carbaldehyde as raw material, in the first organic Reaction in a solvent, dehydration and condensation to obtain compound 1; then place compound 1 in a second organic solvent, add an oxidizing agent to close the ring and dehydrogenate to obtain; wherein: The first organic solvent is one or a combination of two or more selected from toluene, methanol, ethanol, methylene chloride and chloroform; The second organic solvent is one or a combination of two or more selected from benzene, toluene, p-xylene, glacial acetic acid and methylene chloride; The oxidizing agent is palladium carbon, manganese acetate hydrate, lead tetraacetate or 2,3-dichloro-5,6-dicyano-p-benzoquinone. 9. The use of the compound of claim 1 or a pharmaceutically acceptable salt thereof in the preparation of antitumor drugs. 10. The antitumor drug prepared with the compound of claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient.