CN111499608B

CN111499608B - Benzo [ c ] benzopyrone derivatives and application thereof

Info

Publication number: CN111499608B
Application number: CN202010311104.XA
Authority: CN
Inventors: 周本宏; 李妍; 韦平; 兰昱
Original assignee: Wuhan University WHU
Current assignee: Wuhan University WHU
Priority date: 2020-04-20
Filing date: 2020-04-20
Publication date: 2022-04-15
Anticipated expiration: 2040-04-20
Also published as: CN111499608A

Abstract

The invention discloses a benzo [ c ]]Benzopyrone derivatives and their use. Different N-terminal end groups are introduced into the urolithin derivative through a 1, 3-dibromopropane bridge chain, and a series of benzo [ c ] containing Uro B skeleton (shown as formula (I)) is designed and synthesized]Benzopyrone) chemical derivatives. Can be used for preparing various antitumor drugs, especially bladder cancer.

Description

Benzo [ c ] benzopyrone derivatives and application thereof

Technical Field

The invention belongs to the field of medicinal chemistry, and particularly relates to a benzo [ c ] benzopyrone derivative and application thereof in treating cancer diseases.

Background

Bladder cancer is the most common malignancy of the urinary system. The biological behavior of bladder tumor is complex and changeable, and the bladder tumor is easy to recur, invade and transfer. Apart from surgery, systemic chemotherapy has become the standard treatment for metastatic bladder cancer. The chemotherapy drugs commonly used for treating bladder cancer include adriamycin, hydroxycamptothecin, mitomycin and the like, however, because of the wide occurrence of drug resistance, bladder cancer cells are less sensitive to the drugs at present, and the clinical application effect of the existing drugs is poor. Therefore, the development of new anti-bladder cancer drugs is more and more urgent, and the search for low-toxicity and high-efficiency anticancer lead compounds from natural drugs is one of the hot spots of medical research.

Phenolic compounds such as ellagitannins are secondary metabolites of fruits and vegetables, are beneficial to human health, and have been shown to be involved in antioxidant, anti-inflammatory, and potential tumor prevention. In recent years, a great deal of research is carried out on Chinese medicinal materials rich in ellagitannins by a plurality of researchers, and the result shows that the content of the ellagitannins in organisms is low, the ellagitannins cannot reach the concentration of the ellagitannins which can exert the biological activity, and the ellagitannins are mostly further metabolized into ellagic acid and Urolithins (Uro) which are easier to absorb under the action of intestinal flora in the bodies, so the uroritins can be true substances of the ellagitannins which can exert the biological activity in the bodies.

Chemically, Urolithins are benzo [ c ] benzopyrone derivatives with different hydroxyl substitutions. Urolithins is found in plasma mostly in the form of a micromolar concentration of dextran glycoside. The Urolithins skeleton can be regarded as that benzene rings are combined with cis-o-hydroxy cinnamic acid lactone, and the hydroxy groups of Uro B, Uro A, Uro C and Uro D are increased in sequence according to the number of phenolic hydroxy groups. At present, research on the urolithin derivatives is less, and the research mainly focuses on the activity research of the urolithin glucuronic acid derivatives and the modification of coumarin structures similar to the urolithin glucuronic acid derivatives. The glucuronic acid acidification and sulfation forms of the compound are proved to have protective effect on myocardial cells, and the introduction of meta-N-benzyl can be used as a cholinesterase inhibitor, and in addition, the compound has potential application prospect in the treatment of Alzheimer's disease. However, no research on the urolithin skeleton derivatives has been found in the field of anticancer.

Disclosure of Invention

In order to solve the defects in the prior art, the invention provides a benzo [ c ] benzopyrone derivative with a structure shown in a general formula (I) or a pharmaceutically acceptable salt thereof. Different N-terminal end groups are introduced into the Urolithin derivatives through a 1, 3-dibromopropane bridge chain, a series of chemical derivatives containing Uro B skeleton (benzo [ c ] benzopyrone) are designed and synthesized, the core of the design is to improve the water solubility of Urolithin, enhance the bioavailability of Urolithin and obtain stronger biological activity for inhibiting the proliferation of tumor cells.

The invention also aims to provide application of the benzo [ c ] benzopyrone derivative in preparing medicaments for treating cancer diseases.

In order to achieve the purpose, the invention adopts the following technical scheme:

in a first aspect, a benzo [ c ] benzopyrone derivative with a structure of general formula (I) or a derivative thereof is provided

Pharmaceutically acceptable salts:

wherein the content of the first and second substances,

z is substituted or unsubstituted-O (CH2) n-, n is an integer of 2-6, the substituent is hydroxyl or halogen, or a carbon chain in Z contains double bonds or oxygen atoms;

m is 0, 1 or 2;

x is O, N or one of CH;

r is one or more of hydrogen, substituted or unsubstituted C1-5 alkyl, hydroxyl, tert-butyloxycarbonyl and carbonyl.

Preferably, the benzo [ C ] benzopyrone derivative or pharmaceutically acceptable salt with the structure of the general formula (I), wherein the substituted C1-5 alkyl is selected from halogen substituted C1-5 alkyl, the halogen is fluorine, chlorine, bromine and iodine, and the unsubstituted C1-5 alkyl is selected from methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl or isopentyl.

Preferably, the benzo [ c ] benzopyrone derivative or pharmaceutically acceptable salt with the structure of the general formula (I) is selected from any one of the following compounds or pharmaceutically acceptable salts thereof:

3- (3- (4-methylpiperazin-1-yl) -propyl) -6H-benzo [ c ] chromen-6-one (BH 1901);

3- (3-morpholin-1-yl) -propyl) -6H-benzo [ c ] chromen-6-one (BH 1902);

3- (3- (4-methylpiperidin-1-yl) -propyl) -6H-benzo [ c ] chromen-6-one (BH 1903);

3- (3- (pyrrolidinyl-1-yl) -propyl) -6H-benzo [ c ] chromen-6-one (BH 1905).

Preferably, the benzo [ c ] benzopyrone derivative or pharmaceutically acceptable salt with the structure of the general formula (I) is a salt containing a pharmaceutically acceptable anion: any one of hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate or bisulfate, phosphate or acid phosphate, acetate, lactate, citrate, tartrate, maleate, fumarate, methanesulfonate, gluconate, saccharate, benzoate, ethanesulfonate, benzenesulfonate and p-toluenesulfonate.

The general synthetic method of the benzo [ c ] benzopyrone derivative with the structure of the general formula (I) is formed by connecting a benzo [ c ] benzopyrone parent body with a nitrogen-containing structure through a carbon-containing chain,

in a second aspect, there is provided a pharmaceutical composition comprising an amount of a benzo [ c ] benzopyrone derivative or a pharmaceutically acceptable salt thereof having a structure of formula (I) sufficient to exert anticancer activity and a pharmaceutically acceptable excipient.

Preferably, the pharmaceutical composition comprises pharmaceutically acceptable excipients which are inert solid fillers or diluents and sterile aqueous solution or organic solvents.

Benzo [ c ] benzopyrone derivatives having the structure of formula (I) may contain a chiral center and thus may exist in different enantiomeric and diastereomeric forms. The present invention relates to all optical isomers and all stereoisomers of the compounds of general formula (I), as racemic mixtures and in the form of individual enantiomers and diastereomers of such compounds, and to methods of treatment of all pharmaceutical compositions containing or using them, as defined above.

In a third aspect, an application of the benzo [ c ] benzopyrone derivative with the structure of the general formula (I) or the pharmaceutically acceptable salt or the pharmaceutical composition in preparing a medicament for treating cancer diseases is provided.

Preferably, the cancer-like disease is bladder cancer.

Compared with the prior art, the technical scheme of the invention has the following beneficial effects:

CCK-8 experiment results for detecting cancer cell proliferation activity show that the compound hydrochloride has obvious proliferation inhibition effect on human bladder cancer cell T24, and particularly the derivative BH1905 shows the inhibition activity which is optimal to a positive control medicament. Therefore, the compound can be applied to preparing various antitumor drugs, and is particularly suitable for bladder cancer.

Drawings

FIG. 1 is a nuclear magnetic resonance hydrogen spectrum of a compound BH 1901;

FIG. 2 is a NMR spectrum of Compound BH 1902;

FIG. 3 is a hydrogen nuclear magnetic resonance spectrum of compound BH 1903;

FIG. 4 is a hydrogen nuclear magnetic resonance spectrum of compound BH 1905;

Detailed Description

The features and advantages of the present invention will be further understood from the following detailed description taken in conjunction with the accompanying drawings. The examples provided are merely illustrative of the method of the present invention and do not limit the remainder of the disclosure in any way. Example 1 preparation of 3- (3- (4-methylpiperazin-1-yl) -propyl) -6H-benzo [ c ] chromen-6-one (BH1901)

Reaction scheme 1

1) Synthesizing an intermediate: urolithin B (20.2g), 1, 3-dibromopropane (40.4g) and potassium carbonate (27.6g) were weighed in this order in a 500mL round-bottom flask, and heated at 50 ℃ for 2 hours with acetone (200mL) as a solvent. The reaction was stopped when the fluorescent spot of the light blue starting material disappeared by monitoring with Thin Layer Chromatography (TLC). And (3) a spin-drying system, dissolving the solid powder by adding petroleum ether, and performing secondary spin-drying. Silica gel is used as a stationary phase, and petroleum ether: ethyl acetate 1:0, 20:1, 10:1, 8:1, and concentrated to give 14.4g of a white powdery solid. The nuclear magnetic resonance test proves that the intermediate is the target compound, namely the dibromopropylurolithin B intermediate.

2) Synthesizing a target compound: 2.2g of the intermediate, 1.1g of 4-methylpiperazine, 1.1g of potassium carbonate and 20mL of acetonitrile were sequentially added to a 100mL round-bottom flask and refluxed at 60 ℃ for 2 hours. The feeding is carried out according to the molar ratio of 1:1:2, the system exists in the form of suspension during the reaction, the proportion of the fluorescence spots with different polarities is not changed, and the reaction is stopped after the TLC detection reaction is finished. The system was spin dried by repeated addition of acetonitrile, 5.4g of silica gel was added, and the sample was sequentially stir-fried with methanol and dichloromethane as solvents. Silica gel as stationary phase, dichloromethane: methanol was eluted in the forward direction at 1:0, 20:1, 10:1, 0:1 and concentrated to give 0.6g of a viscous yellow-white solid. And re-dissolving with 5ml methanol, separating with thick silica gel plate as stationary phase and methanol as mobile phase, and separating with normal phase thin layer plate. When the blue fluorescent stripes are separated from the dark blue stripes at the bottom, the silica gel on which the blue fluorescent stripes are arranged at the upper part of the thin-layer plate is scraped off, hot methanol is stirred and dissolved at 60 ℃, and the mixture is subjected to suction filtration and spin drying to obtain 0.4g of yellow oily solid. The structure was confirmed by nuclear magnetic resonance, as shown in FIG. 1.

3) Preparation of hydrochloride salt: saturated ethyl acetate hydrochloride solution is prepared firstly, and the principle is that high boiling point acid is used for preparing low boiling point acid. Reacting excessive concentrated sulfuric acid with sodium chloride to generate hydrogen chloride gas and sodium bisulfate, and introducing the hydrogen chloride gas into an ethyl acetate solution to obtain the catalyst. And introducing a compound dissolved by ethyl acetate, oscillating, observing that snowflake-shaped particles are separated out, gradually forming milky floccule precipitates along with the reaction, standing, performing suction filtration to obtain white needle-shaped crystals, and drying to constant weight.

White needle-shaped crystals are obtained. 1H NMR (400MHz, DMSO-d6) δ 8.30(d, J ═ 8.1Hz,1H, Ar-H),8.26 to 8.17(m,2H, Ar-H),7.90(t, J ═ 7.1Hz,1H, Ar-H),7.59(t, J ═ 7.6Hz,1H, Ar-H),7.04 to 6.88(m,2H, Ar-H),4.10(t, J ═ 6.3Hz,2H, -CH2-),2.48 to 2.17(m,10H, -CH2-),2.14(s,3H, N-CH3),1.93 to 1.83(m,2H, -CH2-), anal.calcd for C21H24O3N2, ESI-MS: 352.43[ M + H ] +

Example 2 preparation of 3- (3-morpholin-1-yl) -propyl) -6H-benzo [ c ] chromen-6-one (BH1902)

The title compound was prepared as in example 1, substituting morpholine for 4-methylpiperazine. White crystals are obtained. As shown in fig. 2. 1H NMR (400MHz, DMSO-d6) δ 8.29(d, J ═ 8.1Hz,1H, Ar-H),8.25 to 8.15(m,2H, Ar-H),7.89(t, J ═ 7.6Hz,1H, Ar-H),7.58(t, J ═ 7.5Hz,1H, Ar-H),6.97(dd, J ═ 4.5,2.2Hz,2H, Ar-H),4.10(t, J ═ 6.3Hz,2H, O-CH2),3.57(t, J ═ 4.4Hz,4H, -CH2-O-CH2),2.41(dd, J ═ 18.6,11.4Hz,6H, N-CH2),1.90(p, J ═ 6.6, H, CH2 — cd 20 Hz, C — 5920 Hz, C: 339.38[ M + H ] +.

Example 3 preparation of 3- (3- (4-methylpiperidin-1-yl) -propyl) -6H-benzo [ c ] chromen-6-one (BH1903)

The title compound was prepared by substituting 4-methylpiperazine for 4-methylpiperidine according to the procedure of example 1. Yellow crystals are obtained. As shown in fig. 3. 1H NMR (400MHz, DMSO-d6) δ 8.17 to 8.27(m,3H, Ar-H),7.87(t, J ═ 7.5Hz,1H, Ar-H),7.56(t, J ═ 7.5Hz,1H, Ar-H),6.93(d, J ═ 6.8Hz,2H, Ar-H),4.06(t, J ═ 5.8Hz,2H, -O-CH2-),2.81(d, J ═ 10.9Hz,2H),2.39(t, J ═ 6.9Hz,2H, -CH2-),1.84(t, J ═ 9.6Hz,4H, -CH2-),1.54(d, J ═ 12.0Hz,2H, -CH2-),1.27(s, 1H-), 1H ═ 11, 10H ═ 10H, 2H ═ CH2-),1.54(d, J ═ 12.10H, — 10H, — 11H, 10H ═ 10H, —,2H —: 351.44[ M + H ] +.

Example 4 preparation of 3- (3- (pyrrolidin-1-yl) -propyl) -6H-benzo [ c ] chromen-6-one (BH1905)

The title compound was prepared by the method of example 1, substituting 4-methylpiperazine for pyrrolidine. White crystals are obtained. As shown in fig. 4. 1H NMR (400MHz, DMSO-d6) δ 8.29(d, J ═ 8.1Hz,1H, Ar-H),8.25 to 8.14(m,2H, Ar-H),7.89(t, J ═ 8.2Hz,1H, Ar-H),7.58(t, J ═ 7.6Hz,1H, Ar-H),6.97(dd, J ═ 6.5,2.3Hz,2H, Ar-H),4.10(t, J ═ 6.3Hz,2H, -O-CH2-),2.62(t, J ═ 7.3Hz,2H, -CH2-),2.00 to 1.85(m,6H, -CH2-),1.71(s,4H, -CH2-), anal.esi for C20H21 cd 3 MS, 3N: 323.44[ M + H ] +.

Table 1: number of Compounds prepared in examples and structural formulas thereof

[ example 5 ] in vitro anti-human bladder cancer cell T24 proliferation Activity study of derivatives

Adopts CCK-8[2- (2-methoxy-4-nitrophenyl) -3- (4-nitrophenyl) -5- (2, 4-disulfonic acid benzene) -2H-tetrazole monosodium salt]Method for measuring the Inhibitory Concentration (IC) of a urolithin derivative on T24 cells of human bladder cancer₅₀)。

1) T24 cell culture and passage: the complete culture medium is RPMI-1640 culture medium: fetal bovine serum: qing-streptomycin ═ 90:10: 1; at 37 ℃ with 5% CO₂As shown in FIG. 1, the culture was carried out in an incubator saturated with humidity. When the cell growth density is more than 80%, digesting with trypsin digestion solution containing 0.05% EDTA, observing the cells in suspension state under a microscope, stopping digestion with complete culture medium, centrifuging at 1000r/min for 5 minutes, pouring out supernatant, resuspending the cells with fresh culture medium, and carrying out passage at a ratio of 1: 4.

2) Preparing liquid medicine and grouping administration: the hydrochloride of the compound to be tested in the experimental group is dissolved by a basic culture medium and diluted to different concentration gradients, and 5-fluorouracil is used as a positive control drug. And a blank group and a normal control group are additionally arranged, wherein the blank group is only culture medium liquid with the same amount so as to eliminate the influence of the color of the culture medium on the absorbance, and the normal control group is only added with the culture medium and the cells and is not added with a test medicament.

3) And (3) activity determination: taking T24 cells in logarithmic growth phase, adjusting cell density by a cell counter, and then carrying out cell density adjustment at 1x10 per hole⁵The density of (A) was inoculated in a 96-well plate, 6 multiple wells were set, and 5% CO was performed at 37 ℃₂Culturing in an incubator. After 24 hours of adherent culture, the cell supernatant was discarded, and 200. mu.L of each of 6 derivatives at different concentrations was added to continue the culture. The plate was removed after 24, 36, 48 hours of drug stimulation, the supernatant was discarded, 100. mu.L of serum-free medium containing 5% CCK8 was added, incubation was carried out in an incubator for 1 to 2 hours, and the absorbance value (A) was measured at 450nm with a microplate reader.

Each set of experiments was repeated 3 times. According to the formula: the cell growth inhibition (%) was (normal control a value-experimental a value)/(normal control a value-blank a value) x 100%), and the T24 cell inhibition (%) was calculated. Median Inhibitory Concentration (IC)₅₀) Defined as the drug concentration when 50% of the tumor cells survived. Making standard curve of cell growth inhibition rate according to the measured A value, and obtaining corresponding medicine on the standard curveAnd (4) concentration.

Measured IC₅₀See table 2.

TABLE 2 Activity of the individual Compounds HCl on human bladder cancer cells T24

Claims

1. A benzo [ c ] benzopyrone derivative or a pharmaceutically acceptable salt thereof, characterized by being selected from any one of the following compounds:

3- (3-morpholin-1-yl) -propyl) -6H-benzo [ c ] chromen-6-one (BH 1902);

3- (3- (pyrrolidinyl-1-yl) -propyl) -6H-benzo [ c ] chromen-6-one (BH 1905).

2. The benzo [ c ] benzopyrone derivative or pharmaceutically acceptable salt thereof according to claim 1, wherein the salt is a salt containing a pharmaceutically acceptable anion: any one of hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate or bisulfate, phosphate or acid phosphate, acetate, lactate, citrate, tartrate, maleate, fumarate, methanesulfonate, gluconate, benzoate, ethanesulfonate, benzenesulfonate and p-toluenesulfonate.

3. A pharmaceutical composition comprising a therapeutically effective amount of a benzo [ c ] benzopyrone derivative or a pharmaceutically acceptable salt thereof according to claim 1 or 2 and a pharmaceutically acceptable excipient.

4. The pharmaceutical composition of claim 3, wherein the pharmaceutically acceptable excipient is an inert solid filler or diluent or an organic solvent.

5. Use of a benzo [ c ] benzopyrone derivative or a pharmaceutically acceptable salt thereof according to claim 1 or 2 or a pharmaceutical composition according to claim 3 or 4 for the preparation of a medicament for the treatment of bladder cancer.