CN102126938B - Curcumin analog and preparation method and application thereof - Google Patents

Abstract

The invention discloses a curcumin analog and a preparation method and application thereof, relates to a curcumin analog and provides a curcumin analog. The curcumin analog is red powder, and has the chemical name of 1,7-di(4-hydroxyl-3-allyl benzene)-1,6-heptadiene-3,5-diketone and the molecular weight of 388.1. The molecular structure of curcumin is modified and reformed on the basis of known active compound curcumin, so that a novel compound which is the 1,7-di(4-hydroxyl-3-allyl benzene)-1,6-heptadiene-3,5-diketone with higher antitumor activity is prepared. The curcumin analog remarkably suppresses the growth of nasopharyngeal darcinoma CNE2 cells, human cervical carcinoma Hela cells, human fibrosarcoma HT1080 cells and human liver cancer Bel-7402 cells, and can be widely applied to preparation of antitumor medicaments.

Description

A kind of curcumin analogue and its preparation method and application

technical field

The invention relates to a compound, in particular to a curcumin analogue and its preparation method and application.

Background technique

The chemical structural formula of curcumin is:

Curcumin (curcumin) is a phenolic pigment extracted from the turmeric rhizome of the genus Curcuma longa in the family Zingiberaceae. Its anti-tumor effect was first proposed by Indian scholar Kuttan in 1985 (1, Ramadasan Kuttan, P.Bhanumathy, K.Nirmala et al. al. Cancer Letters (1985) 29: 197-202), and has been widely concerned by scholars at home and abroad. In recent years, in vivo and in vitro experiments have shown that curcumin can change the metabolic detoxification process and activation process of carcinogenic factors, and can change the nature of carcinogenic factors, reduce or remove carcinogenic factors that can enter the intracellular target and key parts, and can inhibit carcinogenic agents. Initiation and promotion of tumorigenesis and development (2. Wang Chunbin. Research progress of curcumin and its application in cardiovascular diseases. Progress in Cardiovascular Diseases, 2005, 26(6), 614-616; 3. Jin Li, Yang Shiyong. Research progress of curcumin. Foreign Medicine Traditional Chinese Medicine, 1997, 19(3): 49-50), and has the advantages of low toxicity, high bioavailability and rich resources. Curcumin can induce cell cycle arrest and apoptosis (4, Huang Dongsheng, Chen Jinhe, Wu Jiliang. Experimental research on apoptosis of human lung cancer cells induced by curcumin. Journal of Xianning Medical College, 2002, 16 (4): 251-255), while It can inhibit the invasion and metastasis of tumors (5, Xiao Xuyang. Research progress on anti-tumor mechanism of curcumin. Journal of Jinzhou Medical College, 2006, 27(1), 56-58), and is a promising anti-tumor drug. The definite antitumor activity of curcumin and its simple structure, easy synthesis and modification provide a good lead for drug development.

At present, in the modification of the structure of curcumin, many valuable conclusions have been obtained, especially the type and position of the substituent on the benzene ring, the β-diketone part, the 4-position methylene substitution, and the structure of the middle connecting chain. The influence and contribution of the pharmacophore, such as stretching and changing, and the planarity of the molecule as a whole, on the pharmacological activity can be used to guide the further research and development of new curcumin drugs.

Contents of the invention

The first object of the present invention is to provide a curcumin analogue.

The second object of the present invention is to provide a preparation method of curcumin analogs.

The third object of the present invention is to provide an application of a curcumin analogue in the preparation of antitumor drugs.

The chemical name of said a kind of curcumin analogue is 1,7-two (4-hydroxyl-3-allyl phenyl)-1,6-heptadiene-3,5-dione, molecular weight is 388.1, The structural formula is:

The curcumin analog is a red powder.

The preparation method of described a kind of curcumin analogue comprises the following steps:

1) Preparation of 4-allyloxybenzaldehyde

Under the protection of nitrogen, add 4-hydroxybenzaldehyde and anhydrous potassium carbonate to the organic solvent to obtain solution A, add allyl bromide to the organic solvent to obtain solution B, then add solution B to solution A, and carry out heating and reflux reaction After the reaction, the organic solvent is removed, the residue is added to a solution C composed of ethyl acetate and saline solution, shaken, the organic layer is separated, and the ethyl acetate is removed after drying to obtain 4-allyloxybenzaldehyde;

2) Preparation of 3-allyl-4-hydroxybenzaldehyde

The 4-allyloxybenzaldehyde obtained in step 1) is subjected to a rearrangement reaction, and the reaction product is separated by silica gel column chromatography to obtain 3-allyl-4-hydroxybenzaldehyde;

3) Preparation of 1,7-bis(4-hydroxyl-3-allylphenyl)-1,6-heptadiene-3,5-dione

Dissolve acetylacetone and boron trioxide in ethyl acetate to obtain solution D, add n-butylamine to ethyl acetate to obtain solution E; tributyl borate and the 3-allyl-4-hydroxyl obtained in step 2) Benzaldehyde was added to ethyl acetate to obtain solution F, solution F was added to solution D to obtain solution G; then solution E was added to solution G to obtain solution H; solution H was acidified with an acid solution, the aqueous layer and the organic layer were separated, and the organic layer was combined. After layering, carry out drying, reduce pressure, remove ethyl acetate, carry out silica gel column chromatography separation to the residue, promptly obtain curcumin analogue: 1,7-bis(4-hydroxyl-3-allylphenyl)- 1,6-Heptadiene-3,5-dione.

In step 1), the organic solvent can be acetone or tetrahydrofuran, etc.; the molar concentration of 4-hydroxybenzaldehyde in the solution A can be 0.05～0.5mmol/L, and the molar concentration of anhydrous potassium carbonate can be 0.1 ～1.0mmol/L, the molar concentration of allyl bromide in the solution B can be 0.1～1.0mmol/L; the temperature of the heating reflux reaction can be 78～80°C, and the time can be 2～5h; The saline solution can be sodium chloride solution or sodium sulfate solution, etc., and the volume ratio of ethyl acetate and saline solution in the solution C can be 2: (1-2).

In step 2), the conditions of the rearrangement reaction are: temperature 160-200° C., time 5-8 hours; the silica gel of the silica gel column chromatography is 200-400 mesh.

In step 3), the molar concentration of acetylacetone in the solution D can be 0.11～1.1mmol/L, the molar concentration of boron trioxide can be 0.07～0.7mmol/L, and the n-butylamine in the solution E Molar concentration can be 0.11～1.1mmol/L; The molar concentration of tributyl borate in the described solution G can be 0.21～2.1mmol/L, and the molar concentration of 3-allyl-4-hydroxybenzaldehyde can be 0.21～ 2.1mmol/L, the volume ratio of solution E and solution G in described solution H can be 1: 1, the volume ratio of described solution H and acid solution can be 5: (1～2), and described acid solution can be sulfuric acid solution or hydrochloric acid solution.

Starting from the known active compound curcumin, the present invention modifies and transforms the molecular structure of curcumin, and prepares a new compound with better antitumor activity: 1,7-di(4-hydroxy-3-allyl Phenyl)-1,6-heptadiene-3,5-dione compounds. It significantly inhibits the growth of nasopharyngeal carcinoma CNE2 cells, human cervical carcinoma HeLa cells, human fibrosarcoma HT1080 cells and human liver cancer Bel-7402 cells, and can be widely used in the preparation of antitumor drugs.

Detailed ways

Example 11, Preparation of 7-two (4-hydroxyl-3-allylphenyl)-1,6-heptadiene-3,5-dione

The preparation method of 1,7-two (4-hydroxy-3-allylphenyl)-1,6-heptadiene-3,5-dione comprises the following steps:

1) Under the protection of nitrogen, take 1.22g of 4-hydroxybenzaldehyde and 2.76g of anhydrous potassium carbonate into 50mL of anhydrous acetone, and stir at room temperature for 30min to obtain a mixed solution;

2) Add 50 mL of acetone solution containing 1.73 mL of allyl bromide to the mixed solution obtained in step 1), and heat to reflux at 80° C. for 3 h until no raw material point can be detected on the silica gel plate. The solvent was removed under reduced pressure to give a concentrated residue. Add 40mL of ethyl acetate solution and 20mL of sodium chloride aqueous solution, and shake. The organic layer was separated, dried over anhydrous magnesium sulfate, filtered, and the solvent was removed under reduced pressure to obtain intermediate product 1: 4-allyloxybenzaldehyde, 1.60 g, yellow oily liquid, yield 98%.

3) Put 1.60 g of 4-allyloxybenzaldehyde into a round bottom flask, install a condenser, rearrange at 200°C for 5 hours, and then cool to room temperature. Then the product was separated by silica gel column chromatography, V (n-hexane): V (ethyl acetate) = 5: 1 as eluent, to obtain intermediate product two: 3-allyl-4-hydroxybenzaldehyde, 1.38g , colorless oily liquid, yield 86%.

4) Dissolve 426 mg of acetylacetone and 209 mg of diboron trioxide into 10 mL of ethyl acetate, and stir at 50° C. for 30 min. Then add 10ml of ethyl acetate solution containing 1.38g of 3-allyl-4-hydroxybenzaldehyde obtained in step 3) and 1.96g of tributyl borate. After stirring for 30min, the ethyl acetate solution containing 0.43mL of n-butylamine 10 mL of the solution was slowly added dropwise to the above reaction solution. The reaction was stirred overnight at 50°C.

5) The reaction mixture was acidified with HCl (0.4N, 6 mL) and shaken. The aqueous and organic layers were separated, and the aqueous layer was extracted three times with ethyl acetate. The organic layer solutions were combined, dried over anhydrous sodium sulfate and filtered. After removing the organic solvent under reduced pressure, the residue was subjected to silica gel column chromatography, V (petroleum ether): V (acetone) = 3: 1 as eluent, to obtain the purified target product: 1,7-bis(4- Hydroxy-3-allylphenyl)-1,6-heptadiene-3,5-dione, 1.12g (2.89mmol), red powder, yield 68%, the total yield of the whole reaction is: 57.8 %. The product was identified by mass spectrometry.

Curcumin was purchased from Sinopharm Chemical Reagent Co., Ltd., and the reagents used in the synthesis were all analytically pure. ¹ H-NMR and ¹³ C-NMR were measured on a Bruker av400 superconducting nuclear magnetic resonance instrument, and were measured in a deuterated organic solvent with TMS as an internal standard. Mass spectra were measured using an Applied Biosystems 3200Q TRAP LC/MS/MS System quadrupole mass spectrometer. Infrared was measured on a Nicolet Avatar 360 infrared absorption spectrometer. UV-spectrometry uses Shimadzu's UV-260spectrometer; microplate reader uses Bio-Rad's products. The test results are as follows:

Spectral data of 1,7-bis(4-hydroxy-3-allylphenyl)-1,6-heptadiene-3,5-dione:

UV (H ₂ O) λ max (log ε) 285 nm (4.03), 471 nm (4.23). ¹ H NMR (400M Hz, DMSO-d ₆ ): δ3.30(d, J=6.4Hz, 4H), 5.04(m, 4H), 5.98(m, 2H), 6.07(s, 1H), 6.62( d, J=15.9Hz, 2H), 6.85(d, J=7.8Hz, 2H), 7.41(brd, J=7.8Hz, 2H), 7.42(brs, 2H), 7.52(d, J=15.9Hz, 2H), 10.09(s, 2H). ¹³ CNMR (400M Hz, DMSO-d ₆ ): 33.64, 100.75, 115.37, 115.62, 120.62, 125.76, 126.80, 128.32, 130.07, 136.61, 140.44, 157.52.MS-183. ESI, m/z 387.4 [MH] ^- .

The obtained product is a kind of curcumin analog, chemical name is 1,7-bis(4-hydroxyl-3-allylphenyl)-1,6-heptadiene-3,5-dione, The molecular weight is 338.1. The structural formula is:

Example 21, the antitumor activity of 7-bis(4-hydroxyl-3-allylphenyl)-1,6-heptadiene-3,5-dione

The present invention studies the antitumor activity of 1,7-bis(4-hydroxy-3-allylphenyl)-1,6-heptadiene-3,5-dione, and finds that it has obvious antitumor activity. Tumor activity, the tumor cells studied include nasopharyngeal carcinoma CNE2 cells, human cervical cancer HeLa cells, human fibrosarcoma HT 1080 cells and human liver cancer Bel-7402 cells. The inhibitory rate of different concentrations of 1,7-bis(4-hydroxy-3-allylphenyl)-1,6-heptadiene-3,5-dione (BHDD) and curcumin (Curcumin) on tumor cells (%) can be found in Table 1.

Table 1

Take the tumor cells in the logarithmic growth phase to make a single-cell suspension, inoculate 1× ¹⁰⁴ cells per well in three 96-well plates, and place them in a cell culture incubator at 37°C with a saturated humidity of 5% _CO2 Cultivate for 24 hours, and when the cells adhere to the wall and grow to 80%, add 1,7-bis(4-hydroxy-3-allylphenyl )-1,6-heptadiene-3,5-dione, without 1,7-bis(4-hydroxy-3-allylphenyl)-1,6-heptadiene-3,5 - Cells in diketone medium served as a negative control and zeroed for blank wells. The cells were placed in a 37°C incubator with saturated humidity containing 1% O ₂ , 5% CO ₂ and 95% N ₂ , and continued to culture for 48 hours. Then measure the absorbance value (A) of each well at 475 nm in a microplate reader. The inhibition rate was calculated according to the following formula: inhibition rate (%)=[1-(average absorbance value of experimental group-average absorbance value of zero-adjusted group)]/(average absorbance value of control group-average absorbance value of zero-adjusted group)×100%.

1) Inhibitory effect of 1,7-bis(4-hydroxy-3-allylphenyl)-1,6-heptadiene-3,5-dione on nasopharyngeal carcinoma CNE2 cells

The antitumor activity of 1,7-bis(4-hydroxy-3-allylphenyl)-1,6-heptadiene-3,5-dione and curcumin on nasopharyngeal carcinoma CNE2 cells showed that: the The compounds all have significant inhibitory effects on the proliferation of nasopharyngeal carcinoma CNE2 cells. When acting at 50,100 μmol/L, compound 1,7-bis(4-hydroxyl-3-allylphenyl)-1,6-heptadiene-3,5-dione showed stronger activity than curcumin Inhibition of the proliferation of nasopharyngeal carcinoma CNE2 cells.

2) Inhibitory effect of 1,7-bis(4-hydroxy-3-allylphenyl)-1,6-heptadiene-3,5-dione on human cervical cancer HeLa cells

The antitumor activity of 1,7-bis(4-hydroxy-3-allylphenyl)-1,6-heptadiene-3,5-dione and curcumin on human cervical cancer HeLa cells showed that: in When acting at 50, 100, and 200 μM, the compound exhibited a stronger inhibitory effect on the proliferation of human cervical cancer HeLa cells than curcumin, and it had a significant concentration-dependent effect.

3) Inhibitory effect of 1,7-di(4-hydroxy-3-allylphenyl)-1,6-heptadiene-3,5-dione on human fibrosarcoma HT 1080 cells

The results of the antitumor activity of 1,7-bis(4-hydroxy-3-allylphenyl)-1,6-heptadiene-3,5-dione and curcumin on human fibrosarcoma HT 1080 cells showed: When acting at 50,100 μmol/L, compound 1,7-bis(4-hydroxyl-3-allylphenyl)-1,6-heptadiene-3,5-dione showed stronger activity than curcumin Inhibition of proliferation of human fibrosarcoma HT 1080 cells.

4) Inhibitory effect of 1,7-bis(4-hydroxy-3-allylphenyl)-1,6-heptadiene-3,5-dione on human liver cancer Bel-7402 cells

The results of the antitumor activity of 1,7-bis(4-hydroxy-3-allylphenyl)-1,6-heptadiene-3,5-dione and curcumin on human liver cancer Bel-7402 cells showed: When acting at 50, 100, and 200 μmol/L, the compound exhibited a stronger inhibitory effect on the proliferation of human liver cancer Bel-7402 cells than curcumin, and it had a significant concentration dependence.

Claims (8)

1. a preparation method of curcumin analogs, characterized in that the chemical name of said curcumin analogs is 1,7-two (4-hydroxyl-3-allylphenyl)-1,6-heptanedi En-3,5-dione, the molecular weight is 388.1, and the structural formula is:  The preparation method comprises the following steps: 1) Preparation of 4-allyloxybenzaldehyde Under the protection of nitrogen, add 4-hydroxybenzaldehyde and anhydrous potassium carbonate to the organic solvent to obtain solution A, add allyl bromide to the organic solvent to obtain solution B, then add solution B to solution A, and carry out heating and reflux reaction After the reaction, the organic solvent is removed, and the residue is added to a solution C composed of ethyl acetate and saline solution, shaken, and the organic layer is separated, and the ethyl acetate is removed after drying to obtain 4-allyloxybenzaldehyde; 2) Preparation of 3-allyl-4-hydroxybenzaldehyde The 4-allyloxybenzaldehyde obtained in step 1) is subjected to a rearrangement reaction, and the reaction product is separated by silica gel column chromatography to obtain 3-allyl-4-hydroxybenzaldehyde; 3) Preparation of 1,7-di(4-hydroxy-3-allylphenyl)-1,6-heptadiene-3,5-dione Dissolve acetylacetone and diboron trioxide in ethyl acetate to obtain solution D, add n-butylamine to ethyl acetate to obtain solution E; tributyl borate and 3-allyl-4-hydroxyl obtained in step 2) Benzaldehyde was added to ethyl acetate to obtain solution F, solution F was added to solution D to obtain solution G; then solution E was added to solution G to obtain solution H; solution H was acidified with an acid solution, the aqueous layer and the organic layer were separated, and the organic layer was combined. After layering, drying, decompression, removal of ethyl acetate, the residue is subjected to silica gel column chromatography to obtain curcumin analogs: 1,7-bis(4-hydroxyl-3-allylphenyl)- 1,6-Heptadiene-3,5-dione. the 2. the preparation method of a kind of curcumin analog as claimed in claim 1 is characterized in that in step 1), described organic solvent is acetone or THF. the 3. the preparation method of a kind of curcumin analogue as claimed in claim 1 is characterized in that in step 1), the molar concentration of 4-hydroxybenzaldehyde in described solution A is 0.05～0.5mmol/L, without The molar concentration of potassium carbonate water is 0.1-1.0 mmol/L, and the molar concentration of allyl bromide in the solution B is 0.1-1.0 mmol/L. the 4. The preparation method of a curcumin analog as claimed in claim 1, characterized in that in step 1), the temperature of the heating-reflux reaction is 78-80°C, and the time is 2-5h. the 5. the preparation method of a kind of curcumin analog as claimed in claim 1, is characterized in that in step 1) in, described saline solution is sodium chloride solution or sodium sulfate solution, ethyl acetate in described solution C The volume ratio with saline solution is 2:1-2. the 6. The preparation method of a curcumin analog as claimed in claim 1, characterized in that in step 2), the conditions of the rearrangement reaction are: temperature 160-200°C, time 5-8h. the 7. The preparation method of a kind of curcumin analogue as claimed in claim 1, is characterized in that in step 2), the silica gel of described silica gel column chromatography is 200～400 meshes. the 8. The preparation method of a kind of curcumin analog as claimed in claim 1, it is characterized in that in step 3), the molar concentration of acetylacetone in the described solution D is 0.11～1.1mmol/L, diboron trioxide The molar concentration of n-butylamine is 0.07～0.7mmol/L, the n-butylamine molar concentration in the described solution E is 0.11～1.1mmol/L; The molar concentration of tributyl borate in the described solution G is 0.21～2.1mmol/L, The molar concentration of 3-allyl-4-hydroxybenzaldehyde is 0.21～2.1mmol/L, and the volume ratio of solution E and solution G in described solution H is 1: 1, and the volume ratio of described solution H and acid solution The ratio is 5:1~2, and the acid solution is sulfuric acid solution or hydrochloric acid solution. the