CN118580215A - A piperate ester compound with anti-tumor activity and its preparation method and application - Google Patents

Abstract

The present invention relates to the technical field of pharmaceutical chemistry, and specifically discloses a piperate ester compound with anti-tumor activity and a preparation method and application thereof, including a compound as shown in formula I or a pharmaceutically acceptable salt thereof: wherein Linker is independently selected from C1-C4 alkane chain, olefin chain, halogen-substituted C1-C4 alkane chain, olefin chain; R ₁ to R ₉ are independently selected from hydrogen, C ₁ -C ₆ alkyl, halogen, cyano, -NO ₂ , acetyl or trifluoromethyl. The compound of formula I of the present invention or its pharmaceutical composition is used in the preparation of a drug for preventing and/or treating tumors. The present invention changes the amide bond of piperine into an ester bond to obtain a novel ester anti-tumor active compound; the compound has a significant inhibitory effect on Hela tumor cells and MDA tumor cells, which is significantly better than piperine, and has a good application value.

Description

A piperate ester compound with anti-tumor activity and its preparation method and application

Technical Field

The invention relates to the technical field of pharmaceutical chemistry, in particular to a piperate ester compound with anti-tumor activity and a preparation method and application thereof.

Background Art

Malignant tumors are a common disease that endangers human life and health. The incidence and mortality of malignant tumors have been on the rise worldwide. At present, the treatment methods for malignant tumors mainly include surgery, radiotherapy and chemotherapy, among which chemotherapy is mainly based on synthetic drugs. The inhibitory effect of chemotherapy drugs on tumors is worthy of recognition. It is also one of the more effective and commonly used treatment methods for malignant tumors, but its toxic side effects are extensive and serious, and there is a problem of drug resistance. In addition, chemotherapy drugs have poor selectivity for tumor cells and normal cells. While killing or inhibiting tumor cells, they can also damage the growth of normal cells, and have a direct impact on the functions of the heart, liver, kidneys and nervous system, and have certain toxicity to the human body. Therefore, it is very necessary to find low-toxic and efficient anti-tumor drugs in tumor treatment. Anti-tumor drugs derived from natural plants have shown unique advantages and broad application prospects due to their low toxicity and high efficiency.

Piperine is a natural amide compound extracted from pepper, with the chemical formula of C ₁₇ H ₁₉ NO ₃ and the chemical name of (E,E)-1-[5-(1,3-benzodioxolan-5-yl)-1-oxo-2,4-pentadienyl]-piperidine.

The properties are colorless, odorless, monoclinic prism crystals with a burning sensation after tasting. The structure of piperine can be mainly divided into three parts: the parent structure aromatic heterocycle-benzodioxolane, α, β-unsaturated fatty hydrocarbon chain, and piperidine ring. Modern pharmacology has found that piperine has pharmacological effects such as cardiovascular protection, enhancing immunity, anti-tumor, improving nervous system diseases, and anti-inflammatory. On December 14, 2021, piperine started a Phase II clinical trial in the United States for the treatment of multiple myeloma, prostate cancer, smoldering multiple myeloma, etc. However, during use, its solubility is poor, absorption is poor, and activity is weak. For this reason, a piperic acid ester compound with anti-tumor activity and its preparation method and application are provided.

Summary of the invention

The purpose of the present invention is to provide a piperate ester compound with anti-tumor activity and a preparation method and application thereof in view of the defects of the prior art, so as to solve the problems raised by the above-mentioned background technology.

To achieve the above object, the present invention provides the following technical solution: a piperate ester compound with anti-tumor activity, comprising a compound as shown in Formula I or a pharmaceutically acceptable salt thereof:

Wherein Linker is independently selected from C1-C4 alkane chain, alkene chain, halogen-substituted C1-C4 alkane chain or alkene chain;

R ₁ to R ₉ are each independently selected from hydrogen, C ₁ -C ₆ alkyl, halogen, cyano, -NO ₂ , acetyl or trifluoromethyl.

As a preferred technical solution of the present invention, it also includes a compound of any of the following general structural formulas or its salt, stereoisomer or hydrate:

As a preferred technical solution of the present invention, the compound is one of the following compounds:

A method for preparing a piperate ester compound with anti-tumor activity, the specific preparation route is as follows:

wherein R ₁ to R ₉ are as defined in the compound of formula I;

The specific steps are as follows:

The compound of formula 1 and the compound of formula 2 are reacted by condensation to obtain the compound of formula I.

As a preferred technical solution of the present invention, the reaction is carried out at 20-50°C;

The reaction solvent is dichloromethane, tetrahydrofuran, N,N-dimethylformamide or acetonitrile;

The base is potassium carbonate, triethylamine, cesium carbonate, DIPEA, NMI or NMM;

The condensing agent is EDCI and HOBT, EDCI and DMAP, HATU, TCFH, COMU or TBTU.

As a preferred technical solution of the present invention, the reaction solvent is dichloromethane or acetonitrile;

The base is triethylamine or potassium carbonate;

The condensing agent is EDCI and HOBT, EDCI and DMAP or HATU.

As a preferred technical solution of the present invention, the molar ratio of the compound of formula I to the compound of formula 2 is 1:0.8 to 1:1.6, and the molar ratio of the compound of formula I to the condensing agent is 1:0.8 to 1:1.6.

As a preferred technical solution of the present invention, the molar ratio of the compound of formula I to the compound of formula 2 is 1:1; the molar ratio of the compound of formula I to the condensing agent is 1:1.2.

A pharmaceutical composition comprises a compound of formula I and a pharmaceutically acceptable carrier.

A use of a compound of formula I or a pharmaceutical composition thereof in the preparation of a drug for preventing and/or treating a tumor, wherein the tumor is cervical cancer.

Compared with the prior art, the present invention has the following beneficial effects:

The present invention has the following significant advantages: (1) the amide bond of piperine is changed into an ester bond to obtain a novel ester anti-tumor active compound; (2) the compound has a significant inhibitory effect on Hela tumor cells and MDA tumor cells, which is significantly better than piperine and has good application value.

DETAILED DESCRIPTION

The preparation method of compound of the present invention is described in more detail below, but these specific preparation methods do not constitute any restriction to the scope of the present invention.In addition, reaction conditions such as reactant, solvent, alkali, the amount of compound used, reaction temperature, reaction time etc. are not limited to the following examples.

The compounds of the present invention can also be conveniently prepared by optionally combining various synthetic methods described in the specification or known in the art. Such a combination can be easily performed by those skilled in the art.

To facilitate the description of the synthetic routes and methods of the examples below, the abbreviations of the main raw materials or reagents used in the examples are summarized in Table 1 below.

Table 1: Abbreviations of main raw materials or reagents used in the examples

Example 1: Preparation of (2E,4E)-5-(benzo[d][1,3]dihydroxy-5-yl)penta-2,4-dienoic acid-3-(1-(dimethylamino)ethyl)phenyl ester:

Step 1: Synthesis of (2E,4E)-5-(benzo[d][1,3]dioxol-5-yl)penta-2,4-dienoic acid:

5g of piperine was dissolved in 20% KOH ethanol solution and heated to reflux at 80°C for 7h. After filtration, the solid compound was washed with anhydrous ethanol and dried to obtain 4g of brown solid compound (5-(3,4-methylenedioxyphenyl)-2,4-pentadienoic acid potassium). Subsequently, the brown solid was directly dissolved in distilled water and diluted hydrochloric acid was added to adjust the pH to 1. The mixture was extracted with ethyl acetate three times and evaporated to dryness to obtain 3g of yellow crystalline compound (2E,4E)-5-(benzo[d][1,3]dioxolane-5-yl)penta-2,4-dienoic acid with a yield of 81%.

Step 2: Synthesis of (2E,4E)-5-(benzo[d][1,3]dihydroxy-5-yl)penta-2,4-dienoic acid-3-(1-(dimethylamino)ethyl)phenyl ester:

3-(1-(dimethylamino)ethyl)phenol (200 mg, 1.210 mmol), (2E,4E)-5-(benzo[d][1,3]dihydroxy-5-yl)penta-2,4-dienoic acid (264.03 mg, 1.210 mmol), EDCI (278.35 mg, 1.452 mmol), HOBT (196.19 mg, 1.452 mmol), triethylamine (504.71 μL, 3.631 mmol), and 3 mL of dichloromethane were added to a 100 mL round-bottom flask in sequence, and the mixture was stirred at room temperature for 2 h. After concentration under reduced pressure, column chromatography (DCM: MeOH = 60:1) was performed to obtain 124 mg of a white solid with a yield of 28.0%. ¹ H NMR (500MHz, DMSO-d6) δ7.58(ddd,J＝15.2,7.4,2.9Hz,1H),7.37(t,J＝7.8Hz,1H),7.28(d,J＝1.5Hz,1H),7.19(d,J＝7.7Hz,1H),7.13–7.08(m,3H),7.06( ¹³ C NMR (126MHz, DMSO-d6) δ165.36,150.94,148.94,148.54,147.57,146.24,142.11,130.80,129.50,125.10,125.06,124.05,120.94,120.59,119.18,10 9.05,108.94,106.31,106.21,102.07,101.95,101.88,64.67,42.93,19.93.

Example 2: Preparation of 3-(1-(dimethylamino)ethyl)phenyl(E)-3-(benzo[d][1,3]dihydroxy-5-yl)acrylate:

3-(1-(dimethylamino)ethyl)phenol (200 mg, 1.210 mmol), (E)-3-(benzo[d][1,3]dihydroxy-5-yl)acrylic acid (232.53 mg, 1.210 mmol), EDCI (278.35 mg, 1.452 mmol), HOBT (196.19 mg, 1.452 mmol), triethylamine (504.71 μL, 3.631 mmol), and 3 mL of dichloromethane were added to a 100 mL round-bottom flask in sequence, and the mixture was stirred at room temperature for 2 h. After concentration under reduced pressure, column chromatography (DCM: MeOH = 60: 1) was performed to obtain 111 mg of a white solid with a yield of 27.0%. ¹ HNMR (500MHz, DMSO-d6) δ7.78(d,J＝15.9Hz,1H),7.51(d,J＝1.5Hz,1H),7.38(t,J＝7.8Hz,1H),7.29(dd,J＝8.1,1.6Hz,1H),7.20(d,J＝7.7Hz,1H),7.11(d,J =1.5Hz,1H),7.07(dd,J＝7.6,1.9Hz,1H),7.00(d,J＝8.0Hz,1H),6.74(d,J＝15.9Hz,1H),6.11(s,2H),3.35(d,J＝6.6Hz,1H),2.12(s,6H),1.29(s,3H) ^1. 3C NMR(126MHz,DMSO-d6)δ165.62,150.96,150.21,148.61,146.75,146.11,129.55,128.83,126.02,125.15,121.00,120.65,115.47,109.02,107.35,10 2.18,64.63,42.89,19.88.

Example 3: Preparation of 3-(1-(dimethylamino)ethyl)benzene 2-(benzo[d][1,3]dioxy-5-yl)acetate:

2-(Benzo[d][1,3]dihydroxy-5-yl)acetic acid (200 mg, 1.11 mmol), EDCI (255 mg, 1.33 mmol), DMAP (160 mg, 1.33 mmol), triethylamine (309 μL, 2.22 mmol), 2 mL N,N-dimethylformamide were added to a 100 mL round-bottom flask in sequence. After reacting at room temperature for 1 h, 3-(1-(S)-(N,N-dimethylamino)ethyl)phenol (200.00 mg, 1.11 mmol) was added and the reaction was continued at room temperature for 4 h. After concentration under reduced pressure, column chromatography (DCM: MeOH = 40: 1) was performed to obtain 170 mg of a transparent oil with a yield of 40.57%. ¹ H NMR (500MHz, DMSO-d6) δ7.34(t,J＝7.8Hz,1H),7.17(dt,J＝7.7,1.3Hz,1H),7.04(t,J＝2.0Hz,1H),6.97(dd,J＝9.3,1.8Hz,2H),6.89(d,J＝7.9Hz,1H),6.83( dd,J＝7.9,1.7Hz,1H),6.01(s,2H),3.86(s,2H),3.28(q,J＝6.7Hz,1H),2.08(s,6H),1.24(d,J＝6.7Hz,3H). ¹³ C NMR (126MHz, DMSO-d6) δ170.14,150.47,147.25,146.25,145.97,129.08,127.47,124.72,122.71,120.31,119.97,109.96,108.14,100.91,64.22,42. 48,19.50.

Example 4: Preparation of 3-(1-(dimethylamino)ethyl)phenylbenzo[d][1,3]dioxolane-5-carboxylate:

3-(1-(dimethylamino)ethyl)phenol (200 mg, 1.210 mmol), benzo[d][1,3]dioxolane-5-carboxylic acid (201.02 mg, 1.210 mmol), EDCI (278.35 mg, 1.452 mmol), HOBT (196.19 mg, 1.452 mmol), triethylamine (504.71 μL, 3.631 mmol), and 3 mL of dichloromethane were added to a 100 mL round-bottom flask in sequence, and reacted at room temperature for 2 h. After reduced pressure concentration, column chromatography (DCM: MeOH = 60: 1) was performed to obtain 105 mg of a white oil with a yield of 27.7%. ¹ H NMR (500MHz, CDCl ₃ ) δ7.84(dd,J＝8.2,1.7Hz,1H),7.63(d,J＝1.6Hz,1H),7.38(t,J＝7.8Hz,1H),7.22(d,J＝7.7Hz,1H),7.19–7.16(m,1H),7.13–7.09(m,1 H), 6.92 (d, J = 8.2Hz, 1H), 6.09 (s, 2H), 3.32 (q, J = 6.7Hz, 1H), 2.24 (s, 6H), 1.40 (d, J = 6.7Hz, 3H). ¹³ C NMR (126MHz, CDCl ₃ )δ151.02,148.06,145.74,129.15,126.08,124.88,123.55,120.77,120.03,109.92,108.14,101.95,77.05,65.55,43.12.

Example 5: Preparation of 3-(1-(dimethylamino)ethyl)phenyl(E)-3-(6-bromobenzo[d][1,3]dihydroxy-5-yl)acrylate:

To a 100 mL round-bottom flask, (E)-3-(6-bromobenzo[d][1,3]dihydroxy-5-yl)acrylic acid (300.00 mg, 1.11 mmol), HATU (507.08 mg, 1.33 mmol), potassium carbonate (460.77 mg, 3.33 mmol) were added in sequence, dissolved in 6 mL of acetonitrile and stirred in an ice bath for 30 minutes, and finally 3-(1-(dimethylamino)ethyl)phenol (201.99 mg, 1.22 mmol) was added, and the mixture was reacted at 25°C for 4 hours. The mixture was concentrated under reduced pressure and column chromatography (eluted with ethyl acetate) was used to obtain 150.24 mg of an oily compound with a yield of 33.4%. ¹ H NMR (500MHz, DMSO-d6) δ8.02(d,J＝15.8Hz,1H),7.71(s,1H),7.48(t,J＝7.9Hz,1H),7.38(s,1H),7.35–7.30(m,1H),7.28(s,1H),7.24–7.18(m,1H),6 .88(d,J＝15.7Hz,1H),6.18(s,2H),4.03(q,J＝7.1Hz,1H),2.39(s,6H),1.45(d,J＝6.8Hz,3H). ¹³ CNMR(126MHz,DMSO-d6)δ170.79,165.27,151.13,150.95,148.57,144.17,130.10,126.80,126.14,121.89,118.43,118.35,113.27,107.59,103.24,6 4.57,60.22,41.74,40.59,40.50,40.42,40.33,40.26,40.16,40.09,40.00,39.92,39.83,39.66,39.50,38.71,21.22,18.07,14.55.

Example 6: Preparation of 3-(1-(dimethylamino)ethyl)phenyl 2,2-difluorobenzo[d][1,3]dioxolane-5-carboxylate:

2,2-Difluorobenzo[d][1,3]dioxolane-5-carboxylic acid (300.00 mg, 1.31 mmol), HATU (600.38 mg, 1.58 mmol), potassium carbonate (545.55 mg, 3.94 mmol) were added to a 100 mL round-bottom flask in sequence, dissolved in 6 mL acetonitrile and stirred in an ice bath for 30 minutes, and finally 3-(1-(dimethylamino)ethyl)phenol (239.15 mg, 1.44 mmol) was added, and the mixture was reacted at 25°C for 4 hours. The mixture was concentrated under reduced pressure and column chromatography (DCM:MeOH=40:1) was used to obtain 210.15 mg of a yellow solid with a yield of 45.91%. ¹ H NMR (500MHz, CDCl ₃ ) δ8.05(dd,J＝8.4,1.7Hz,1H),7.89(d,J＝1.7Hz,1H),7.38(t,J＝7.9Hz,1H),7.24(dt,J＝7.8,1.3Hz,1H),7.20(t,J＝2.0Hz,1H),7.18( d, J＝8.4Hz, 1H), 7.11 (ddd, J＝8.1, 2.4, 1.0Hz, 1H), 3.33 (q, J＝6.7Hz, 1H), 2.25 (s, 6H), 1.40 (d, J＝6.7Hz, 3H). ¹³ C NMR (126MHz, CDCl ₃ )δ163.62,150.78,147.42,146.12,143.82,133.77,131.73,129.68,129.27,127.20,125.87,125.24,120.50,120.07,111.20,109.34,77.34,77 .09,76.83,65.54,43.06,31.92,30.29,29.69,26.89,22.68,19.90,14.09.

Compound activity test on cancer cells:

To explore the effects of compounds on the activity of cancer cells, the MTT method was used to detect cytotoxicity.

The specific test method is as follows:

Cell culture: Human cancer cell line Hela cells, human breast cancer cells MDA-MB-231 and human normal kidney embryonic 293T cells were purchased from Beijing National Biomedical Laboratory and cultured in DMEM (KGM12800-500) or MEM medium (KGM41500-500) containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin, in a 5% CO ₂ , 37°C constant temperature incubator (Thermo Fisher Scientific, BB150). When the cell confluence reached 70%-80%, 0.25% trypsin was added for digestion, resuspending and culturing. Cells in the logarithmic growth phase and in good growth state were selected for research.

Methyl thiazolyl tetrazolium (MTT) was used to determine cell viability. The hemocytometer method was used for cell counting, and in all experiments, cell viability was greater than 95%. 293T and Hela cells were seeded in 96-well plates at 1×10 ⁴ cells per ml. Different concentrations of drugs (0-50 μM) dissolved in 100 μL of culture medium (containing 1% FBS) were added to each well and incubated for 48 h. Using a plate centrifuge (5 min, 2000 rpm), the supernatant was discarded, 10 μL MTT (5 mg/mL) solution was added to each well, incubated at 37°C for 4 h, the plate was centrifuged and the supernatant was discarded, 100 μL DMSO was added to each well, and the plate was placed on a shaker for 10 min to fully dissolve the formazan crystals. The absorbance was measured at a wavelength of 570 nm using a microplate reader (BioTek, USA). Cytotoxicity was determined compared with the control group (DMSO). The concentration of drug inducing 50% inhibition of cell growth ( _IC50 ) was determined by nonlinear regression using the curve fitting algorithm of GraphPad Prism 9 (GraphPad software, La Jolla, CA, USA).

To explore the effect of the compound on the activity of tumor cells, the MTT method was used to detect cytotoxicity. As shown in Table 1, the effect of the compound on the activity of different tumor cells was calculated, and the half inhibitory concentration IC ₅₀ (48h) of the drug on each cell line was calculated. The results showed that the inhibitory effect of the compound on Hela tumor cells and MDA tumor cells was significantly better than that of piperine, and it has good development potential.

Table 2: Inhibitory activity of compounds against different tumor cells

The above embodiments only express the implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention.

Claims (10)

1. A piperate ester compound having anti-tumor activity, characterized in that it comprises a compound as shown in Formula I or a pharmaceutically acceptable salt thereof: Wherein Linker is independently selected from C1-C4 alkane chain, alkene chain, halogen-substituted C1-C4 alkane chain or alkene chain; R ₁ to R ₉ are each independently selected from hydrogen, C ₁ -C ₆ alkyl, halogen, cyano, -NO ₂ , acetyl or trifluoromethyl. 2. The piperate ester compound with anti-tumor activity according to claim 1, characterized in that it also comprises a compound of any of the following general structural formulas or its salt, stereoisomer or hydrate: 3. The piperate ester compound with anti-tumor activity according to claim 2, characterized in that the compound is one of the following compounds: 4. A method for preparing a piperate ester compound with anti-tumor activity as claimed in any one of claims 1 to 3, characterized in that the specific preparation route is as follows: wherein R ₁ to R ₉ are as defined in the compound of formula I; The specific steps are as follows: The compound of formula 1 and the compound of formula 2 are reacted by condensation to obtain the compound of formula I. 5. The method for preparing a piperate ester compound with anti-tumor activity according to claim 4, characterized in that the reaction is carried out at 20-50° C.; The reaction solvent is dichloromethane, tetrahydrofuran, N,N-dimethylformamide or acetonitrile; The base is potassium carbonate, triethylamine, cesium carbonate, DIPEA, NMI or NMM; The condensing agent is EDCI and HOBT, EDCI and DMAP, HATU, TCFH, COMU or TBTU. 6. The method for preparing a piperate ester compound with anti-tumor activity according to claim 4, characterized in that the reaction solvent is dichloromethane or acetonitrile; The base is triethylamine or potassium carbonate; The condensing agent is EDCI and HOBT, EDCI and DMAP or HATU. 7. The method for preparing a piperate ester compound with antitumor activity according to claim 4, characterized in that the molar ratio of the compound of formula 1 to the compound of formula 2 is 1:0.8 to 1:1.6, and the molar ratio of the compound of formula 1 to the condensing agent is 1:0.8 to 1:1.6. 8. The method for preparing a piperate ester compound with anti-tumor activity according to claim 7, characterized in that the molar ratio of the compound of formula 1 to the compound of formula 2 is 1:1; and the molar ratio of the compound of formula 1 to the condensing agent is 1:1.2. 9. A pharmaceutical composition, characterized in that it comprises the compound of formula I as claimed in claim 1 and a pharmaceutically acceptable carrier. 10. Use of the compound of formula I according to claim 1 or the pharmaceutical composition according to claim 9 in the preparation of a medicament for preventing and/or treating a tumor, wherein the tumor is cervical cancer.