CN108752306B

CN108752306B - 6-fluoro-1, 3-dihydroxy xanthone and preparation method and application thereof

Info

Publication number: CN108752306B
Application number: CN201810795885.7A
Authority: CN
Inventors: 周北斗; 王欣; 翁智敏; 阮志鹏
Original assignee: Putian University
Current assignee: Putian University
Priority date: 2018-07-19
Filing date: 2018-07-19
Publication date: 2021-09-17
Anticipated expiration: 2038-07-19
Also published as: CN108752306A

Abstract

The inventor provides a novel fluoro 1, 3-dihydroxy xanthone compound 6-fluoro-1, 3-dihydroxy xanthone, a preparation method thereof and application thereof in antitumor drugs, in particular to drug preparation in leukemia, lung cancer, liver cancer, breast cancer and colon cancer.

Description

6-fluoro-1, 3-dihydroxy xanthone and preparation method and application thereof

Technical Field

The invention relates to the field of medical chemistry, in particular to 6-fluoro-1, 3-dihydroxy xanthone and a preparation method and application thereof.

Background

Xanthones, also known as xanthones, are the first secondary metabolites isolated from some plants and microorganisms, and contain the parent structure of dibenzo-gamma-pyrones. Xanthone compounds with different structures often have different pharmacological actions or activity degrees. Changes in one or more substituents of the xanthone parent ring can affect its biological activity. The xanthones which have been found to date have a number of important biological activities. Such as antihypertensive, antibacterial, antithrombotic, antitumor, and obesity-suppressing activities.

On the basis of natural medicines, the synthesis of new compounds by chemical means is an important direction for the development of new medicines at present.

Disclosure of Invention

The inventor provides a novel fluoro 1, 3-dihydroxy xanthone compound 6-fluoro-1, 3-dihydroxy xanthone, and a preparation method and application thereof. The structure of the compound is shown as (I):

。

the inventor further provides a preparation method of 6-fluoro-1, 3-dihydroxy xanthone, which comprises the process of reacting 4-fluoro-2-hydroxybenzoic acid, phloroglucinol, phosphorus pentoxide and methane sulfonic acid.

Further, the molar ratio of the 4-fluoro-2-hydroxybenzoic acid to the phloroglucinol is 4-5: 4-5.

Further, the method comprises the following steps:

preparing a mixed solution: dissolving phosphorus pentoxide in methanesulfonic acid to prepare a mixed solution; the dissolving temperature is controlled at 105-115 ℃;

and (3) cyclization reaction: adding 4-fluoro-2-hydroxybenzoic acid and phloroglucinol into the mixed solution to carry out cyclization reaction to generate 6-fluoro-1, 3-dihydroxy xanthone, wherein the reaction temperature is controlled at 88-92 ℃, and the reaction time is controlled at 18-22 min;

solid-liquid separation and purification: and (3) precipitating, filtering and carrying out column chromatography separation on the solution containing the 6-fluoro-1, 3-dihydroxy xanthone after the cyclization reaction is finished to obtain a finished product of the 6-fluoro-1, 3-dihydroxy xanthone.

The inventor also provides application of the 6-fluoro-1, 3-dihydroxy xanthone in preparing antitumor drugs.

Further, the tumor comprises: hematopoietic stem cell tumor, liver cell tumor, lung cell tumor, breast cell tumor, colon cell tumor.

The inventor further provides an anti-tumor medicament which contains the 6-fluoro-1, 3-dihydroxy xanthone or the pharmaceutically acceptable salt or ester thereof as an active ingredient.

The inventor further provides an anti-tumor pharmaceutical composition, which contains the 6-fluoro-1, 3-dihydroxy xanthone or the pharmaceutically acceptable salt or ester thereof as an active ingredient.

Further, the medicine and the medicine composition comprise tablets, capsules, pills, injections, sustained-release preparations and various microparticle administration systems.

Different from the prior art, the technical scheme provides a novel fluoro 1, 3-dihydroxy xanthone compound, a preparation method thereof and application thereof in antitumor drugs, in particular to drug preparation in leukemia, lung cancer, liver cancer, breast cancer and colon cancer.

Detailed Description

In order to explain technical contents, structural features, and objects and effects of the technical means in detail, the following detailed description is given with reference to specific embodiments.

In this embodiment, the chemical structural formula of 6-fluoro-1, 3-dihydroxyxanthone is as follows:

example 1: synthesis of 6-fluoro-1, 3-dihydroxyxanthone

Phosphorus pentoxide (0.68g, 4.8mmol) and 15mL methanesulfonic acid (CH)₃SO₃H) Then, the mixture was added to a 50mL round-bottom flask, heated to 110 ℃ and stirred to dissolve the resulting mixture, and the reaction mixture was cooled to 90 ℃ and then added with 4-fluoro-2-hydroxybenzoic acid (1.6mmol) and phloroglucinol (0.20g, 1.6mmol) to react at 90 ℃ for 20 min. Pouring the reaction liquid into water, separating out a solid, performing suction filtration and natural drying, and enabling the primary product to pass through a 200-mesh and 300-mesh pressurized silica gel column by using a mixed solvent of ethyl acetate and petroleum ether to obtain the 6-fluoro-1, 3-dihydroxy xanthone which is an orange red solid and has the yield of 56 percent.

The infrared spectrum data are as follows: IR (KBr) v_max 843,1169,1267,1450,1613,1656,3454cm^-1；¹H NMR(500MHz,CD₃SOCD₃)δ6.22(d,1H,J＝2.1Hz,H-2),6.39(d,1H,J＝2.1Hz,H-4),7.33(td,1H,J＝2.4,8.8Hz,H-7),7.55(dd,1H,J＝2.4,9.8Hz,H-5),8.18(dd,1H,J＝6.5,8.8Hz,H-8),11.16(s,OH-3),12.70(s,OH-1)；¹³C NMR(125MHz,CD₃SOCD₃)δ94.2,98.4,102.0,104.7,112.9,117.1,128.2,156.7,157.6,165.0,166.0,167.0,179.0；HR-ESI-MS:m/z 245.0256[M-H]^-(calcd for C₁₃H₆FO₄,245.0256).

The 6-fluoro-1, 3-dihydroxyxanthone can be synthesized according to the following process:

EXAMPLE 26 Synthesis of fluoro-1, 3-dihydroxyxanthone

Phosphorus pentoxide (5mmol) and 15mL of methanesulfonic acid (CH)₃SO₃H) Sequentially adding into a 50mL round-bottom flask, heating to 105 deg.C, stirring to dissolve, cooling the reaction solution to 92 deg.C, adding 4-fluoro-2-hydroxybenzoic acid (1.6mmol) and phloroglucinol (2.0mmol), and reacting at 92 deg.C for 18 min. Pouring the reaction liquid into water, separating out solid, carrying out suction filtration and natural drying, and enabling the primary product to pass through a 200-mesh and 300-mesh pressurized silica gel column by using a mixed solvent of ethyl acetate and petroleum ether to obtain the 6-fluoro-1, 3-dihydroxy xanthone which is an orange red solid.

EXAMPLE 36 Synthesis of fluoro-1, 3-dihydroxyxanthone

Phosphorus pentoxide (4.5mmol) and 15mL of methanesulfonic acid (CH)₃SO₃H) Sequentially adding into a 50mL round-bottom flask, heating to 115 deg.C, stirring to dissolve, cooling the reaction solution to 88 deg.C, adding 4-fluoro-2-hydroxybenzoic acid (2.0mmol) and phloroglucinol (1.6mmol), and reacting at 88 deg.C for 22 min. Pouring the reaction liquid into water, separating out solid, carrying out suction filtration and natural drying, and enabling the primary product to pass through a 200-mesh and 300-mesh pressurized silica gel column by using a mixed solvent of ethyl acetate and petroleum ether to obtain the 6-fluoro-1, 3-dihydroxy xanthone which is an orange red solid.

Example 4: antitumor Activity test of 6-fluoro-1, 3-Dihydroxyxanthone

(1) The MTS method is used for detecting the cell activity principle: MTS is a novel MTT analogue, known as 3- (4, 5-dimethylthiozol-2-yl) -5 (3-carboxymethyloxyphenyl) -2- (4-sulfop heny) -2H-tetrazolium, and is a yellow dye. Succinate dehydrogenase in the mitochondria of living cells can metabolize and reduce MTS to generate soluble Formazan (Formazan) compounds, and the content of the Formazan can be measured at 490nm by using an enzyme labeling instrument. Since the formazan production amount is generally proportional to the number of living cells, the number of living cells can be estimated from the optical density OD value.

(2) Experimental methods

Inoculating cells: preparing single cell suspension by using culture solution (DMEM or RMPI1640) containing 10% fetal bovine serum, inoculating 3000-15000 cells in each well into a 96-well plate, wherein each well volume is 100 mu l, and the adherent cells are inoculated and cultured 12-24h in advance.

Adding a solution of a compound to be detected: compounds were dissolved in DMSO and compounds were prescreened at a concentration of 40. mu.M, with a final volume of 200. mu.l per well, with 3 replicates per treatment.

③ developing color: after culturing for 48h at 37 ℃, removing culture solution in each hole of the adherent cells, and adding 20 mul of MTS solution and 100 mul of culture solution into each hole; discarding 100 mul of culture supernatant from the suspension cells, and adding 20 mul of MTS solution into each well; 3 blank double wells (mixed solution of 20. mu.l MTS solution and 100. mu.l culture medium) were set, and incubation was continued for 2-4 hours to allow the reaction to proceed sufficiently, and then the light absorption value was measured.

And fourthly, color comparison: selecting 492nm wavelength, reading the light absorption value of each hole by a multifunctional microplate reader (MULTISKAN FC), recording the result, and drawing a tumor cell inhibition rate graph by taking the compound number as the abscissa and the cell inhibition rate as the ordinate after data processing.

The human tumor and normal cells detected are as follows: leukemia HL-60, lung cancer A-549, liver cancer SMMC-7721, breast cancer MCF-7, colon cancer SW480, liver cancer Huh-7, liver cancer HepG2, liver cancer MHCC97H, liver cancer PLC/PRF/5, liver cancer SK-HEP-1 and human normal liver cell HL-7702.

Sixthly, for the tumor cells with the inhibition rate of more than 50 percent, determining the IC of the compound on the tumor cells₅₀The value is obtained. Two positive compounds of cisplatin (DDP) and paclitaxel (Taxol) are set in each experiment, a cell growth curve is drawn by taking the concentration as the abscissa and the cell survival rate as the ordinate, and the IC of the compound is calculated by using a two-point method (Reed and Muench method)₅₀The value is obtained.

Using cisplatin (DDP) and paclitaxel (Taxol) as positive control compounds, a small amount of 6-fluoro-1, 3-dihydroxyxanthone synthesized in example 1 was dissolved in DMSO by MTS method to prepare a 40. mu.M solution, and then the tumor cells were examined for inhibitory activity (tables 1 to 3).

Inhibitory Activity of the Compounds of Table 1 against 5 tumor cells

Table 1 The inhibitory activities of compound against five tumor cells

TABLE 2 inhibitory Activity of the Compounds on 6 other hepatocytes

Table 2 The inhibitory activities of compound against the other six liver cells

IC of Table 3 Compounds on liver cancer SMMC-7721 cells₅₀Value of

Table 3 The IC₅₀values of the compound against liver tumor SMMC-7721 cells

From tables 1-2, it can be derived: the inhibition rate of 40 mu M6-fluoro-1, 3-dihydroxy xanthone on leukemia HL-60, lung cancer A-549, breast cancer MCF-7, colon cancer SW480, liver cancer Huh-7, liver cancer HepG2, liver cancer PLC/PRF/5, liver cancer SK-HEP-1 and human normal liver cell HL-7702 is 10-40%. Thus, 6-fluoro-1, 3-dihydroxyxanthone has some inhibitory activity against these cells.

It can also be derived from tables 1-3: the inhibition rate of 40 μ M6-fluoro-1, 3-dihydroxy xanthone on human liver cancer SMMC-7721 is 63.56 + -0.60%, corresponding IC₅₀The value was 23.53. + -. 0.36. mu.M. Therefore, the 6-fluoro-1, 3-dihydroxy xanthone has better inhibitory activity on human liver cancer SMMC-7721.

The 6-fluoro-1, 3-dihydroxy xanthone is a new compound, the synthesis and the biological activity of which are not reported in documents or patents, and can be used for preparing antitumor drugs.

It is noted that, herein, relational terms such as first and second, and the like may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or terminal that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or terminal. Without further limitation, an element defined by the phrases "comprising … …" or "comprising … …" does not exclude the presence of additional elements in a process, method, article, or terminal that comprises the element. Further, herein, "greater than," "less than," "more than," and the like are understood to exclude the present numbers; the terms "above", "below", "within" and the like are to be understood as including the number.

It should be noted that, although the above embodiments have been described herein, the invention is not limited thereto. Therefore, based on the innovative concepts of the present invention, the technical solutions of the present invention can be directly or indirectly applied to other related technical fields by making changes and modifications to the embodiments described herein or by using equivalent structures or equivalent processes performed in the present specification, and are included in the scope of the present invention.

Claims

1.6-fluoro-1, 3-dihydroxyxanthone having the chemical structure shown in formula (I):

Ⅰ

application in preparing anti-leukemia HL-60 medicine.