CN116283961A - A kind of 4-(3,4,5-trimethoxyphenyl)-imidazo[4,5-c]pyridine compound and its application - Google Patents

Abstract

The invention relates to a 4- (3, 4, 5-trimethoxyphenyl) -imidazo [4,5-c]Pyridine compounds, the chemical structure of which is shown in the following formula (I); in the formula (I), R ¹ Is 1-methyl-indol-4-yl, 1-methyl-indol-5-yl or 1-methyl-indol-6-yl. The compound has remarkable inhibition 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, and can be used for preparing antitumor drugs.

Description

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

technical field

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

Background technique

Microtubules are long tubular structures composed of 13 tubulin fibrils. Tubulin fibrils are composed of tubulin subunits. Each tubulin subunit is a dimer formed by the association 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 mitosis process of cancer cells by inhibiting the polymerization of tubulin or the depolymerization of microtubules, interrupting cell mitosis, arresting the cell cycle in G2/M phase, and leading to apoptosis of cancer cells.

All currently marketed tubulin inhibitors bind to either the paclitaxel site or the vinca alkaloid site. Although these inhibitors are effective for various cancers, their clinical application faces some problems, including: not being able to take it orally, high toxicity and side effects, and multidrug resistance. Tubulin inhibitors targeting the colchicine-binding site have the following advantages over marketed tubulin inhibitors. ① Overcome drug resistance of paclitaxel and vinblastine: not only different binding sites, but also not a substrate of multidrug resistance/MDR; ②High oral bioavailability: relatively simple structure, good physical and chemical properties/water solubility and pharmacokinetics Kinetic properties; ③Toxicity and side effects are relatively small: high water solubility, no need to use co-solvents, and less side effects.

In view of the critical role of tubulin in the proliferation and division of cancer cells, and currently there is no marketed drug targeting the colchicine binding site. Designing and developing novel tubulin inhibitors that can overcome paclitaxel resistance, have high oral bioavailability, and have good pharmacokinetic properties targeting the microcolchicine binding site are of great significance and broad scope as anticancer drugs. application prospects.

Contents of the invention

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

The scheme that the present invention solves the problems of the technologies described above is as follows:

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

In formula (I), R ₁ is 1-methyl-indol-4-yl, 1-methyl-indol-5-yl or 1-methyl-indol-6-yl.

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

When R ₁ is 1-methyl-indol-4-yl, the chemistry of the 4-(3,4,5-trimethoxyphenyl)-imidazo[4,5-c]pyridine compounds The structure is:

When R ₁ is 1-methyl-indol-5-yl, the chemistry of the 4-(3,4,5-trimethoxyphenyl)-imidazo[4,5-c]pyridine compounds The structure is:

When R ₁ is 1-methyl-indol-6-yl, the chemistry of the 4-(3,4,5-trimethoxyphenyl)-imidazo[4,5-c]pyridine compounds The structure is:

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

(1) Dissolve 2-bromopyridine-3,4-diamine (A) in ethanol, add 1.2 molar times of 1-methyl-1H- Indolealdehyde (B), 0.15 mole times of p-toluenesulfonic acid, reflux reaction for 24 hours, after the reaction solution was concentrated under reduced pressure, water was added, extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, and column chromatography Separation and purification to obtain intermediate C;

The chemical reaction formula of above-mentioned steps (1) is as follows:

(2) Take the intermediate C obtained in step (1) and dissolve it in ethylene dioxane-water (9:1), and add 1.2 molar times of 3,4,5-trimethoxy according to the molar amount of intermediate C Phenylboronic acid (D), 2.0 mole times of sodium carbonate, catalyzed by 1,1'-bis(di-tert-butylphosphino)ferrocene dichloride palladium, reacted at 80°C for 12 hours under nitrogen protection, and the reaction solution After concentration under reduced pressure, water was added, extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, separated and purified by column chromatography to obtain the 4-(3,4,5-trimethoxyphenyl)- Imidazo[4,5-c]pyridine compounds (I);

The chemical reaction formula of above-mentioned steps (2) is as follows:

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

The 4-(3,4,5-trimethoxyphenyl)-imidazo[4,5-c]pyridine compound of the present invention has the effect on human breast cancer cell MCF-7, human cervical cancer cell HeLa, human liver cancer cell The inhibitory effect of HepG-2 and mouse melanoma cell B16-F10 is remarkable, and can be used for preparing antitumor drugs.

The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments.

Description of drawings

Fig. 1 is 2-(1-methyl-indol-4-yl)-4-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-c] of the present invention Proton NMR spectrum of pyridine (Ia).

Fig. 2 is 2-(1-methyl-indol-4-yl)-4-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-c] of the present invention Carbon NMR spectrum of pyridine (Ia).

Fig. 3 is 2-(1-methyl-indol-5-yl)-4-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-c] of the present invention Proton NMR Spectrum of Pyridine (Ib)

Fig. 4 is 2-(1-methyl-indol-5-yl)-4-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-c] of the present invention Carbon NMR spectrum of pyridine (Ib).

Fig. 5 is 2-(1-methyl-indol-6-yl)-4-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-c] of the present invention Proton NMR spectrum of pyridine (Ic).

Figure 6 shows the 2-(1-methyl-indol-6-yl)-4-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-c] of the present invention Carbon NMR spectrum of pyridine (Ic).

Detailed ways

Embodiment 1 (preparation Ia)

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

The preparation method of compound shown in above-mentioned formula (Ia) is made up of the following steps:

(1) Step 1: Preparation of Compound C1

的化合物：Prepare the structural formula as follows

compound of:

Dissolve (100mg, 0.532mmol) 2-bromopyridine-3,4-diamine (A) in 5mL of absolute ethanol, add (101.6mg, 0.638mmol) 1-methylindole-4-aldehyde (B1), (13.7mg, 0.080mmol) p-toluenesulfonic acid, reflux for 24h. The reaction was monitored by TLC. After the reaction was completed, the reaction solution was concentrated, the concentrate was extracted with ethyl acetate-water, the ethyl acetate layer was dried over anhydrous sodium sulfate, concentrated, and 143 mg of solid was obtained by petroleum ether-ethyl acetate 2:1 column chromatography. Yield 82.2%. The obtained orange solid powder was identified by proton nuclear magnetic resonance spectroscopy, and the result was: ¹ H 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).

(2) Step 2: Preparation of Compound Ia

的化合物：Prepare the structural formula as follows

compound of:

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

The obtained yellow solid powder was identified by proton nuclear magnetic resonance spectrum and carbon nuclear magnetic resonance spectrum, and the identification result was: 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 . 71.

Embodiment 2 (preparation 1b)

The 2-(1-methyl-indol-5-yl)-4-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-c]pyridine ( 1b) the structural formula is:

The compound shown in above-mentioned formula (Ib) is with 2-bromopyridine-3,4-diamine (A), 1-methylindole-5-aldehyde (B2), 3,4,5-trimethoxyphenylboronic acid ( D) as a raw material, the specific preparation method is the same as in Example 1, and the chemical reaction formula is as follows:

The product obtained by the above method was identified by nuclear magnetic resonance spectrum, 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, 152.61,145.38,140.61,138.50,133.60,131.28.08,120.. 35, 120.21, 119.54, 110.54, 106.15, 105.72, 101.48, 60.14, 55.81, 32.68.

Embodiment 3 (preparation Ic)

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

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

The product obtained by the above method was identified by nuclear magnetic resonance spectrum, 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,1H),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.

Embodiment 4 (antitumor activity research)

The in vitro antitumor activity of the compound of the present invention is proved by the following method test. These effects indicate that the compound of the present invention can be used in the treatment of cancer, especially in the treatment of solid tumors, such as human breast cancer, human cervical cancer, human liver cancer and mouse melanoma. The specific test method is as follows:

(1) Cells in the logarithmic growth phase were seeded in a 96-well plate at a density of 5×10 ⁴ cells/mL and 100 μL/well. Culture in a 37°C, 5% CO ₂ , saturated humidity incubator until the cells adhere to the wall.

(2) Aspirate the original medium, and add different concentrations of compounds to each group, the compound concentrations are 0.001 μM, 0.003 μM, 0.01 μM, 0.03 μM, 0.1 μM, 1 μM. 0.1% DMSO was used as the control group, and the culture was continued for 48 hours in the cell culture incubator.

(3) Add 10 μL of MTT solution to each well and incubate in the incubator for 4 hours.

(4) Discard the medium, add 150 μL DMSO to each well, and shake for 10 minutes to fully dissolve the formazan crystals.

(5) Measure the absorbance value at 570 nm with an enzyme-linked immunosorbent assay.

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

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

As: Absorbance of experimental wells (including cells, MTT, compounds)

Ac: Absorbance of control wells (with cells, MTT, no compound)

Ab: Absorbance of blank wells (without cells and compounds, with MTT)

The measured activity results are shown in Table 1:

Table 1 Comparison of antitumor activity (Mean ± SD)

The positive reference substance in the above table is the compound 7 disclosed in Example 1 of the CN106794174A patent application (the publication number is the 0357th paragraph of the CN106794174A patent application). The synthetic route of the compound 7 is as follows:

The synthetic method of compound 7 is the same as Example 1 in the CN106794174A patent application (see [0351] and [0352] of CN106794174A):

(1) Reflux indole-5-aldehyde, 2-bromopyridine-3,4-diamine (3mmol), p-toluenesulfonic acid (0.3mmol), and 15mL of EtOH under argon protection for 24h. Concentrate under reduced pressure to remove the solvent, add 25 mL of water, and extract the mixture with ethyl acetate (3×50 mL). The combined organic layers were dried over MgSO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography to give the desired 4-bromo-2-(1H-indol-5-yl)-1H-imidazo[4,5-c]pyridine.

(2) The corresponding bromide (1eq), 3,4,5-trimethoxyphenylboronic acid (1eq), sodium carbonate (2eq), tetrakistriphenylphosphine palladium (0.1eq) obtained in the above step (1) ) was dissolved in THF (3ml)/water (0.3ml) 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 MgSO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography to give the desired 2-(1H-indol-5-yl)-4-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-c ] Pyridine.

2-(1H-indol-5-yl)-4-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-c]pyridine. ¹ H NMR (400MHz, CDCl ₃ ) 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).

对上述癌细胞的抑制效果均明显优于阳性对照品和对照药品(秋水仙碱)。The above in vitro experimental results show that 4-(3,4,5-trimethoxyphenyl)-imidazo[4,5-c]pyridine compounds have the effect on human breast cancer cell MCF-7, human cervical cancer cell HeLa, human Liver cancer cells HepG-2 and mouse melanoma cells B16-F10 have a strong inhibitory effect, and the compound

The inhibitory effects on the above-mentioned cancer cells are significantly better than the positive control substance and the control drug (colchicine).

Claims (4)

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

In formula (I), R ₁ is 1-methyl-indol-4-yl, 1-methyl-indol-5-yl or 1-methyl-indol-6-yl. 2. A kind of 4-(3,4,5-trimethoxyphenyl)-imidazo[4,5-c]pyridine compound according to claim 1, characterized in that, the described 4 -(3,4,5-trimethoxyphenyl)-imidazo[4,5-c]pyridine compound is one of the following compounds: When R ₁ is 1-methyl-indol-4-yl, the chemistry of the 4-(3,4,5-trimethoxyphenyl)-imidazo[4,5-c]pyridine compounds The structure is:

When R ₁ is 1-methyl-indol-5-yl, the chemistry of the 4-(3,4,5-trimethoxyphenyl)-imidazo[4,5-c]pyridine compounds The structure is:

When R ₁ is 1-methyl-indol-6-yl, the chemistry of the 4-(3,4,5-trimethoxyphenyl)-imidazo[4,5-c]pyridine compounds The structure is:

3. The application of the 4-(3,4,5-trimethoxyphenyl)-imidazo[4,5-c]pyridine compound according to claim 1 or 2 in the preparation of antitumor drugs. 4. application according to claim 3, is characterized in that, described antitumor drug is made of 4-(3,4,5-trimethoxyphenyl)-imidazo[4,5 -c] composed of pyridine compounds and medically acceptable auxiliary materials.

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