A 4-(3,4,5-trimethoxyphenyl)-imidazo[4,5-c]pyridine compound and its application

Abstract

The present invention relates to a 4-(3,4,5-trimethoxyphenyl)-imidazo[4,5-c]pyridine compound, the chemical structure of which is shown in the following formula (I); in the formula (I), R 1 The compound of the present invention has significant inhibitory effects on human breast cancer cells MCF-7, human cervical cancer cells HeLa, human liver cancer cells HepG-2 and mouse melanoma cells B16-F10, and can be used to prepare anti-tumor drugs. #imgabs0#

Description

Technical Field

[0001] The invention relates to 4-(3,4,5-trimethoxyphenyl)-imidazo[4,5-c]pyridine compounds. The compounds can inhibit tumor cell proliferation and can be used for treating tumors. Background Art

[0002] Microtubules are long tubular structures composed of 13 tubulin protofilaments. Tubulin protofilaments are composed of tubulin subunits. Each tubulin subunit is a dimer formed by the combination of two similar globular proteins (α-tubulin and β-tubulin). The growth of cancer cells is highly dependent on the dynamic instability of tubulin. Given that microtubules play a key role in the proliferation and division of cancer cells, tubulin has become an ideal target for anticancer drugs. Tubulin inhibitors interfere with the mitotic process of cancer cells by inhibiting the polymerization of tubulin or the depolymerization of microtubules, interrupting cell mitosis and causing the cell cycle to stagnate at the G2 / M phase, leading to apoptosis of cancer cells.

[0003] All currently marketed tubulin inhibitors bind to the paclitaxel site or the vinca alkaloid site. Although these inhibitors are effective against various cancers, their clinical application faces some problems, including: they cannot be taken orally, have large toxic and side effects, and multidrug resistance. Tubulin inhibitors targeting the colchicine binding site have the following advantages over the marketed tubulin inhibitors. ① Overcoming the drug resistance of paclitaxel and vinca alkaloid: not only do they have different binding sites, but they are also not substrates for multidrug resistance / MDR; ② High oral bioavailability: relatively simple structure, good physicochemical properties / water solubility and pharmacokinetic properties; ③ Relatively small toxic and side effects: high water solubility, no need for cosolvents, and few side reactions.

[0004] Given the key role of tubulin in cancer cell proliferation and division, and the fact that there is currently no marketed drug targeting the colchicine binding site, designing and developing new tubulin inhibitors targeting the colchicine binding site that can overcome paclitaxel resistance, have high oral bioavailability, and have good pharmacokinetic properties as anticancer drugs has important significance and broad application prospects. Summary of the invention

[0005] The technical problem to be solved by the present invention is to provide a 4-(3,4,5-trimethoxyphenyl)-imidazo[4,5-c]pyridine compound, which has a significant inhibitory effect on human breast cancer cells MCF-7, human cervical cancer cells HeLa, human liver cancer cells HepG-2 and mouse melanoma cells B16-F10.

[0006] The solution of the present invention to solve the above technical problems is as follows:

[0007] A 4-(3,4,5-trimethoxyphenyl)-imidazo[4,5-c]pyridine compound, the chemical structure of which is shown in the following formula (I):

[0008]

[0009] In formula (I), R1 is 1-methyl-indol-4-yl, 1-methyl-indol-5-yl or 1-methyl-indol-6-yl.

[0010] The 4-(3,4,5-trimethoxyphenyl)-imidazo[4,5-c]pyridine compound of the present invention is preferably one of the following compounds:

[0011] When R1 is 1-methyl-indol-4-yl, the chemical structure of the 4-(3,4,5-trimethoxyphenyl)-imidazo[4,5-c]pyridine compound is:

[0012]

[0013] When R1 is 1-methyl-indol-5-yl, the chemical structure of the 4-(3,4,5-trimethoxyphenyl)-imidazo[4,5-c]pyridine compound is:

[0014]

[0015] When R1 is 1-methyl-indol-6-yl, the chemical structure of the 4-(3,4,5-trimethoxyphenyl)-imidazo[4,5-c]pyridine compound is:

[0016]

[0017] The preparation method of the above-mentioned 4-(3,4,5-trimethoxyphenyl)-imidazo[4,5-c]pyridine compound comprises the following steps:

[0018] (1) 2-bromopyridine-3,4-diamine (A) is dissolved in ethanol, and 1.2 molar times of 1-methyl-1H-indolealdehyde (B) and 0.15 molar times of p-toluenesulfonic acid are added according to the molar amount of 2-bromopyridine-3,4-diamine, and refluxed for 24 hours. After the reaction solution is concentrated under reduced pressure, water is added, and extraction is performed with ethyl acetate. The organic phase is dried over anhydrous sodium sulfate, and separated and purified by column chromatography to obtain intermediate C;

[0019] The chemical reaction formula of the above step (1) is as follows:

[0020]

[0021] (2) the intermediate C obtained in step (1) is dissolved in ethylenedioxane-water (9:1), 1.2 molar times of 3,4,5-trimethoxyphenylboronic acid (D) and 2.0 molar times of sodium carbonate are added according to the molar amount of the intermediate C, and the mixture is reacted at 80° C. for 12 hours under nitrogen protection under the catalysis of 1,1'-bis(di-tert-butylphosphino)ferrocenepalladium dichloride, and the reaction solution is concentrated under reduced pressure, and water is added, and the mixture is extracted with ethyl acetate. The organic phase is dried over anhydrous sodium sulfate, and separated and purified by column chromatography to obtain the 4-(3,4,5-trimethoxyphenyl)-imidazo[4,5-c]pyridine compound (I);

[0022] The chemical reaction formula of the above step (2) is as follows:

[0023]

[0024] The process route of the above-mentioned method for synthesizing 4-(3,4,5-trimethoxyphenyl)-imidazo[4,5-c]pyridine compounds is as follows:

[0025]

[0026] The 4-(3,4,5-trimethoxyphenyl)-imidazo[4,5-c]pyridine compounds of the present invention have significant inhibitory effects on human breast cancer cells MCF-7, human cervical cancer cells HeLa, human liver cancer cells HepG-2 and mouse melanoma cells B16-F10, and can be used to prepare anti-tumor drugs.

[0027] The present invention is further described below in conjunction with the accompanying drawings and specific embodiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 It is the hydrogen nuclear magnetic resonance spectrum of 2-(1-methyl-indol-4-yl)-4-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-c]pyridine (Ia) described in the present invention.

[0029] Figure 2 The carbon nuclear magnetic resonance spectrum of 2-(1-methyl-indol-4-yl)-4-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-c]pyridine (Ia) of the present invention is shown.

[0030] Figure 3 The hydrogen nuclear magnetic resonance spectrum of 2-(1-methyl-indol-5-yl)-4-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-c]pyridine (Ib) of the present invention is

[0031] Figure 4The carbon nuclear magnetic resonance spectrum of 2-(1-methyl-indol-5-yl)-4-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-c]pyridine (Ib) of the present invention.

[0032] Figure 5 It is the hydrogen nuclear magnetic resonance spectrum of 2-(1-methyl-indol-6-yl)-4-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-c]pyridine (Ic) described in the present invention.

[0033] Figure 6 The carbon nuclear magnetic resonance spectrum of 2-(1-methyl-indol-6-yl)-4-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-c]pyridine (Ic) described in the present invention. DETAILED DESCRIPTION

[0034] Example 1 (Preparation of Ia)

[0035] The structural formula of 2-(1-methyl-indol-4-yl)-4-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-c]pyridine (Ia) prepared in this example is:

[0036]

[0037] The preparation method of the compound represented by the above formula (Ia) comprises the following steps:

[0038] (1) Step 1: Preparation of Compound C1

[0039] The structure is prepared as follows Compounds:

[0040]

[0041] Take (100 mg, 0.532 mmol) 2-bromopyridine-3,4-diamine (A) and dissolve it in 5 mL of anhydrous ethanol, add (101.6 mg, 0.638 mmol) 1-methylindole-4-aldehyde (B1), (13.7 mg, 0.080 mmol) p-toluenesulfonic acid, and reflux for 24 hours. Monitor the reaction by TLC. After the reaction is completed, concentrate the reaction solution, extract the concentrate with ethyl acetate-water, dry the ethyl acetate layer with anhydrous sodium sulfate, concentrate, and obtain 143 mg of solid by petroleum ether-ethyl acetate 2:1 column chromatography, with a yield of 82.2%. The obtained orange solid powder was identified by nuclear magnetic resonance hydrogen spectrum method, and the result was: 1H NMR (400MHz, DMSO) δ8.11(d,J=5.4Hz,1H),7.86(d,J=7.4Hz,1H),7.68(d,J=8.1Hz,1H),7.59(d ,J=5.4Hz,1H),7.55(d,J=2.9Hz,1H),7.47(d,J=2.8Hz,1H),7.36(t,J=7.8Hz,1H),3.88(s,3H).

[0042] (2) Step 2: Preparation of Compound Ia

[0043] The structure is prepared as follows Compounds:

[0044]

[0045] Take (140 mg, 0.428 mmol) of compound C1 and dissolve it in 9.0 mL of dioxane and 1.0 mL of water, add (108.9 mg, 0.513 mmol) of 3,4,5-trimethoxyphenylboronic acid (D), (90.7 mg, 0.856 mmol) of sodium carbonate, (10 mg) of 1,1'-bis(di-tert-butylphosphino)ferrocenepalladium dichloride, and react at 80°C for 12 hours under nitrogen protection. Monitor the reaction by TLC, and concentrate the reaction solution after the reaction is completed. The concentrate is extracted with ethyl acetate-water, and the ethyl acetate layer is dried over anhydrous sodium sulfate and concentrated. 112 mg of solid is obtained by column chromatography with petroleum ether-ethyl acetate 2:1, with a yield of 63.2%.

[0046] The obtained yellow solid powder was identified by H NMR and C NMR, and the identification results were: 1 HNMR (400MHz, DMSO) δ8.41(d,J=3.1Hz,3H),7.89(d,J=7.4Hz,1H),7.68(d,J=8.1Hz,1H),7.60(d,J=2.7Hz,1H ),7.56(d,J=2.9Hz,1H),7.52(d,J=5.3Hz,1H),7.38(t,J=7.8Hz,1H),3.97(s,6H),3.89(s,3H),3.78(s,3H). 13 C NMR (101MHz, DMSO) δ153.16,152.69,145.82,140.96,140.63,138.64,138.32,137.26,133.63, 131.15,125.99,120.93,120.51,118.92,112.45,106.09,105.83,101.85,60.16,55.87,32.71.

[0047] Example 2 (Preparation of Ib)

[0048] The structural formula of 2-(1-methyl-indol-5-yl)-4-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-c]pyridine (Ib) prepared in this example is:

[0049] The compound represented by the above formula (Ib) uses 2-bromopyridine-3,4-diamine (A), 1-methylindole-5-aldehyde (B2), and 3,4,5-trimethoxyphenylboronic acid (D) as raw materials. The specific preparation method is the same as that in Example 1. The chemical reaction formula is as follows:

[0050]

[0051] The product obtained by the above method was identified by nuclear magnetic resonance spectroscopy, and the result was: 1 H NMR (400MHz, DMSO) δ8.49(s,1H),8.41(s,2H),8.36(d,J=5.3Hz,1H),8.14(d,J=8.5Hz,1H),7.67(d ,J=8.6Hz,1H),7.47(t,J=4.5Hz,2H),6.62(d,J=2.9Hz,1H),3.96(s,6H),3.86(s,3H),3.77(s,3H). 13 C NMR (101MHz, DMSO) δ154.07,152.61,145.38,141.23,140.61,138.50,137.58,133.60,131. 27,128.08,120.35,120.21,119.54,110.54,106.15,105.72,101.48,60.14,55.81,32.68.

[0052] Example 3 (Preparation Ic)

[0053] The structural formula of 2-(1-methyl-indol-6-yl)-4-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-c]pyridine (Ic) prepared in this example is:

[0054] The compound represented by the above formula (Ic) uses 2-bromopyridine-3,4-diamine (A), 1-methylindole-5-aldehyde (B3), and 3,4,5-trimethoxyphenylboronic acid (D) as raw materials. The specific preparation method is the same as that in Example 1. The chemical reaction formula is as follows:

[0055]

[0056] The product obtained by the above method was identified by nuclear magnetic resonance spectroscopy, and the result was: 1 H NMR (400MHz, DMSO) δ8.45(s,2H),8.41(s,1H),8.38(d,J=5.3Hz,1H),7.99(d,J=8.2Hz,1H),7.74(d,J=8.3Hz,1 H),7.53(d,J=3.0Hz,1H),7.50(d,J=5.3Hz,1H),6.53(d,J=2.9Hz,1H),3.98(s,6H),3.91(s,3H),3.77(s,3H). 13 C NMR(101MHz,DMSO)δ153.92,152.62,145.45,141.21,140.73,138.46,138.15,136.35,133.51, 132.32,129.94,122.03,120.90,117.81,108.75,106.10,105.78,100.83,60.13,55.75,32.54.

[0057] Example 4 (Study on Antitumor Activity)

[0058] The in vitro anti-tumor activity of the compounds of the present invention is demonstrated by the following test methods. These effects indicate that the compounds of the present invention can be used to treat cancer, especially solid tumors, such as human breast cancer, human cervical cancer, human liver cancer and mouse melanoma. The specific test methods are as follows:

[0059] (1) Cells in the logarithmic growth phase were cultured at a rate of 5×10 4 The cells were seeded in a 96-well plate at a density of 100 μL / well and cultured in a 37°C, 5% CO2, saturated humidity incubator until the cells adhered to the wall.

[0060] (2) The original culture medium was aspirated, and different concentrations of compounds were added to each group, with the concentrations of the compounds being 0.001 μM, 0.003 μM, 0.01 μM, 0.03 μM, 0.1 μM, and 1 μM, respectively. A 0.1% DMSO control group was used, and the cells were cultured in a cell culture incubator for 48 h.

[0061] (3) Add 10 μL MTT solution to each well and incubate in the incubator for 4 h.

[0062] (4) Discard the culture medium, add 150 μL DMSO to each well, and shake for 10 min to fully dissolve the formazan crystals.

[0063] (5) Measure the absorbance at 570 nm using an enzyme-linked immunosorbent assay (ELISA).

[0064] (6) Calculate the cell growth inhibition rate according to the following formula:

[0065] Inhibition rate = [(As-Ab) / (Ac-Ab)] × 100%

[0066] As: absorbance of the experimental well (containing cells, MTT, compounds)

[0067] Ac: Absorbance of control well (containing cells, MTT, no compound)

[0068] Ab: absorbance of blank wells (without cells and compounds, with MTT)

[0069] The measured activity results are shown in Table 1:

[0070] Table 1 Comparison of antitumor activity (Mean±SD)

[0071]

[0072] The positive control in the above table is compound 7 disclosed in Example 1 of patent application with publication number CN106794174A (paragraph 0357 of patent application with publication number CN106794174A). The synthesis route of compound 7 is as follows:

[0073]

[0074] The synthesis method of compound 7 is the same as that of Example 1 in the patent application with publication number CN106794174A (see

[0351] and

[0352] of CN106794174A):

[0075] (1) Indole-5-aldehyde, 2-bromopyridine-3,4-diamine (3mmol), p-toluenesulfonic acid (0.3mmol), and 15mL of EtOH were refluxed under argon protection for 24h. The solvent was removed by concentration under reduced pressure, 25mL of water was added, and the mixture was extracted with ethyl acetate (3×50mL). The combined organic layer was dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified by column chromatography to obtain the desired 4-bromo-2-(1H-indol-5-yl)-1H-imidazo[4,5-c]pyridine.

[0076] (2) The corresponding bromide (1 eq), 3,4,5-trimethoxyphenylboronic acid (1 eq), sodium carbonate (2 eq), and tetrakistriphenylphosphine palladium (0.1 eq) obtained in the above step (1) were dissolved in THF (3 ml) / water (0.3 ml) and refluxed overnight. The solvent was removed by concentration under reduced pressure, and after adding water to the concentrate, it was extracted with ethyl acetate. The organic layer was dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified by column chromatography to obtain the desired 2-(1H-indol-5-yl)-4-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-c]pyridine.

[0077] 2-(1H-indol-5-yl)-4-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-c]pyridine. 1 H NMR (400MHz, CDCl3) δ8.51-8.44(m,2H),8.36(s,1H),8.00(d,J=8.5Hz,1H), 7.54-7.50(m,2H),7.35-7.30(m,2H),6.65(s,1H),4.01(s,6H),3.93(s,3H).

[0078] The above in vitro experimental results show that 4-(3,4,5-trimethoxyphenyl)-imidazo[4,5-c]pyridine compounds have a strong inhibitory effect on human breast cancer cells MCF-7, human cervical cancer cells HeLa, human liver cancer cells HepG-2 and mouse melanoma cells B16-F10. The inhibitory effects on the above-mentioned cancer cells are significantly better than those of the positive control substance and the control drug (colchicine).

Claims

1. A use of a 4-(3,4,5-trimethoxyphenyl)-imidazo[4,5-c]pyridine compound in the preparation of an anti-liver cancer drug, characterized in that: The structural formula of the 4-(3,4,5-trimethoxyphenyl)-imidazo[4,5-c]pyridine compound is: or 2. The use according to claim 1, characterized in that: The anti-liver cancer drug is composed of the 4-(3,4,5-trimethoxyphenyl)-imidazo[4,5-c]pyridine compound and medically acceptable auxiliary materials.

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