CN107698550A - A kind of xanthone compound of 2,4 difluorophenyl substitution and its preparation method and application - Google Patents

Abstract

A 2,4-difluorophenyl-substituted xanthone compound, which is 7-(2,4-difluorophenyl)-1,3-dihydroxyxanthone, its structure is shown in the following formula : 7-(2,4-difluorophenyl)-1,3-dihydroxyxanthone is a new xanthone compound with obvious antitumor activity and tyrosinase inhibitory activity, It can be used to prepare anti-tumor drugs and drugs, health care products, food and cosmetics for regulating tyrosinase. The preparation method of 7-(2,4-difluorophenyl)-1,3-dihydroxyxanthone provided by the present invention has simple operation, mild reaction conditions and high yield, and can be used for 7-(2, Large-scale preparation of 4-difluorophenyl)-1,3-dihydroxyxanthone.

Description

A kind of 2,4-difluorophenyl substituted xanthone compound and its preparation method and application

technical field

The present invention relates to the field of biochemical medicine, in particular to a 2,4-difluorophenyl-substituted xanthone compound and its preparation method and application.

Background technique

Xanthone, also known as xanthone, is a secondary metabolite isolated from some plants and microorganisms, containing the parent structure of dibenzo-γ-pyrone. These compounds have many important biological activities, such as antihypertensive, anti-dysplasia, antibacterial, antithrombotic and antitumor activities. These different biological activities are determined by different substituents on the parent ring of xanthone.

The different substituents on the parent ring of xanthone determine its various biological activities, and the change of one or more substituents will affect its biological activity. In order to obtain a xanthone compound with a certain biological activity, it is necessary to synthesize the xanthone compound first, and then perform relevant activity screening.

Contents of the invention

For this reason, the object of the present invention is to provide a kind of xanthone compound with certain physiological and pharmacological activities.

For realizing the purpose of the present invention, the present invention adopts following technical scheme:

A 2,4-difluorophenyl-substituted xanthone compound, the compound is 7-(2,4-difluorophenyl)-1,3-dihydroxyxanthone, the structure of which is as follows Shown in chemical formula (I):

The present invention also provides a preparation method of the compound shown in formula I, comprising 5-(2,4-difluorophenyl)-2-hydroxybenzoic acid, phloroglucinol, phosphorus pentoxide and methanesulfonic acid the process of reacting.

Preferably, the molar ratio of 5-(2,4-difluorophenyl)-2-hydroxybenzoic acid to phloroglucinol is 1:1.

Preferably, the following steps are included:

1) dissolving phosphorus pentoxide in methanesulfonic acid to prepare the first mixed solution;

2) Add 5-(2,4-difluorophenyl)-2-hydroxybenzoic acid and phloroglucinol to the first mixed solution for ring closure reaction to generate 7-(2,4-difluorophenyl) -1,3-Dihydroxyxanthone;

Preferably, the following steps are also included:

2) After the reaction is completed, the solution containing 7-(2,4-difluorophenyl)-1,3-dihydroxyxanthone is precipitated, filtered by suction, and separated by column chromatography to obtain purified 7-( 2,4-difluorophenyl)-1,3-dihydroxyxanthone.

Preferably, in the step 1), the dissolution temperature is controlled at 105-115°C.

Preferably, in the step 2), the reaction temperature is controlled at 88-92°C, and the reaction time is controlled at 18-22 minutes.

During preparation, temperature control is crucial. If the temperature is too high, the raw materials will be carbonized and the yield will be reduced, while if the temperature is too low, the reaction rate will be affected.

The present invention also provides the use of the compound represented by formula I in inhibiting tyrosinase activity.

The present invention also provides the application of the compound represented by formula I in antitumor drugs.

The present invention also provides the application of the compound represented by formula I in tyrosinase metabolism regulating medicine, health care products, food and cosmetics.

The compound 7-(2,4-difluorophenyl)-1,3-dihydroxyxanthone shown in the formula I provided by the present invention is a new xanthone compound with obvious anti- The tumor activity and the activity of inhibiting tyrosinase can be used to prepare antitumor drugs and tyrosine metabolism regulating drugs, health care products, food and cosmetics. The preparation method of the compound represented by the formula I of the present invention has simple operation, mild reaction conditions and high yield, and can be used for mass preparation of the compound represented by the formula I.

detailed description

In order to describe in detail the technical content, structural features, achieved purpose and effect of the technical solution, it will be described in detail below in conjunction with specific embodiments.

For realizing the purpose of the present invention, the present invention adopts following technical scheme:

A 2,4-difluorophenyl-substituted xanthone compound, the compound is 7-(2,4-difluorophenyl)-1,3-dihydroxyxanthone, the structure of which is as follows Shown in chemical formula (I),

The present invention also provides a preparation method of the compound represented by formula I, comprising the process of reacting 4-trifluoromethyl-2-hydroxybenzoic acid, phloroglucinol, phosphorus pentoxide and methanesulfonic acid.

Preferably, the molar ratio of 4-trifluoromethyl-2-hydroxybenzoic acid to phloroglucinol is 1:1.

The present invention also provides a preparation method of the compound shown in formula I, comprising 5-(2,4-difluorophenyl)-2-hydroxybenzoic acid, phloroglucinol, phosphorus pentoxide and methanesulfonic acid the process of reacting.

Preferably, the molar ratio of 5-(2,4-difluorophenyl)-2-hydroxybenzoic acid to phloroglucinol is 1:1.

Preferably, the following steps are included:

1) dissolving phosphorus pentoxide in methanesulfonic acid to prepare the first mixed solution;

2) Add 5-(2,4-difluorophenyl)-2-hydroxybenzoic acid and phloroglucinol to the first mixed solution for ring closure reaction to generate 7-(2,4-difluorophenyl) -1,3-Dihydroxyxanthone;

Preferably, the following steps are also included: 2) the solution containing 7-(2,4-difluorophenyl)-1,3-dihydroxyxanthone after the completion of the reaction is subjected to precipitation, suction filtration, and column chromatography Separation afforded purified 7-(2,4-difluorophenyl)-1,3-dihydroxyxanthone.

Preferably, in the step 1), the dissolution temperature is controlled at 105-115°C.

Preferably, in the step 2), the reaction temperature is controlled at 88-92°C, and the reaction time is controlled at 18-22 minutes.

The present invention also provides the use of the compound represented by formula I in inhibiting tyrosinase activity.

The present invention also provides the application of the compound represented by formula I in antitumor drugs.

The present invention also provides the application of the compound represented by formula I in tyrosinase metabolism regulating medicine, health care products, food and cosmetics.

The compound 7-(2,4-difluorophenyl)-1,3-dihydroxyxanthone shown in the formula I provided by the present invention is a new xanthone compound with obvious antitumor The activity and the activity of inhibiting tyrosinase can be used for preparing antitumor drugs and drugs for regulating tyrosine metabolism. The preparation method of the compound represented by the formula I of the present invention has simple operation and mild reaction conditions, and can be used for mass preparation of the compound represented by the formula I.

In order to further understand the present invention, the present invention will be described in detail below in conjunction with examples.

Example 1: Synthesis of 7-(2,4-difluorophenyl)-1,3-dihydroxyxanthone

Add 0.68g of phosphorus pentoxide (4.8mmol) and 15mL of methanesulfonic acid (MeSO ₃ H) into a 50mL round-bottomed flask successively, heat to 110°C, stir to dissolve it, then drop the reaction solution to 90°C, and add 0.40 g 5-(2,4-difluorophenyl)-2-hydroxybenzoic acid (1.6mmol) and 0.20g phloroglucinol (1.6mmol) were reacted at 90°C for 20min. Pour the reaction solution into water to precipitate a solid, filter it with suction, and dry it naturally. Use a mixed solvent of ethyl acetate and petroleum ether (V _{ethyl acetate} : V _{petroleum ether} = 1:10) to pressurize the initial product through 200-300 mesh. A silica gel column was used to obtain 0.41 g of a yellow solid with a yield of 76.0%.

¹ H _NMR ⁽ CD ₃ COCD ₃ , 500MHz)δ6.29(1H,d,J=2.2Hz,H-2),6.48(1H,d,J=2.2Hz,H-4),7.20(2H,m,H-3 ',H-5'),7.66(1H,d,J＝8.7Hz,H-5),7.70(1H,m,H-6'),8.02(1H,dq,J＝0.6,1.8,8.7Hz , H-6), 8.32 (1H, t, J=1.8Hz, H-8), 10.3 (1H, s, HO-3), 12.8 (1H, s, HO-1); ¹³ C NMR (CD ₃ COCD ₃ ,125MHz)δ95.0,99.2,103.7,105.0,105.2,105.4, 112.8,119.0,126.3,132.8,136.6,156.3,164.7,166.8,180.9; MS-ESI, m/z 339[M-H] ; HRMS (ESI) m/z _calcd for _C19H9F2O4 (MH): _339.0474 , found: _339.0465 .

Example 2: Antitumor activity test of 6-trifluoromethyl-1,3-dihydroxyxanthone

(1) Principle of MTS assay for cell viability

MTS is a brand new MTT analog, its full name is 3-(4,5-dimethylthiazol-2-yl)-5(3-carboxymethoxyphenyl)-2-(4-sulfopheny)-2H-tetrazolium, it is a yellow color of dyes. Succinate dehydrogenase in the mitochondria of living cells can metabolize and reduce MTS to generate soluble formazan (Formazan) compounds, and the content of formazan can be measured at 490nm with a microplate reader. Under normal circumstances, the amount of formazan produced is directly proportional to the number of living cells, so the number of living cells can be inferred from the optical density OD value.

(2) Experimental method

① Cell inoculation: Prepare a single cell suspension with culture medium (DMEM or RMPI1640) containing 10% fetal bovine serum, inoculate 3,000-15,000 cells per well into a 96-well plate with a volume of 100 μl per well, and inoculate the adherent cells in advance. 12 ~ 24h inoculation culture.

②Add the compound solution to be tested: the compound is dissolved in DMSO, the compound is screened at a concentration of 40 μM/L, the final volume of each well is 200 μl, and 3 replicate wells are set for each treatment.

③Color development: After culturing at 37°C for 48 hours, discard the culture medium in the well for adherent cells, add 20 μl of MTS solution and 100 μl of culture medium for each well; discard 100 μl of culture supernatant for suspension cells, and add 20 μl of MTS solution for each well; set 3 blanks Duplicate wells (a mixture of 20 μl of MTS solution and 100 μl of culture medium), and continue to incubate for 2 to 4 hours to allow the reaction to fully proceed before measuring the light absorption value.

④Colorimetry: Select a wavelength of 492nm, read the light absorption value of each well with a multi-functional microplate reader (MULTISKAN FC), record the results, after data processing, use the compound number as the abscissa, and the cell inhibition rate as the ordinate to draw the inhibition rate of tumor cells picture.

⑤ Nine types of human tumor cells detected are as follows: leukemia HL-60, T cell leukemia MT-4, lung cancer A-549, liver cancer SMMC-7721, liver cancer HepG2, liver cancer Huh-7, colon cancer SW 480, breast cancer MCF-7 and cervical cancer Hela cells.

⑥ For tumor cells with an inhibition rate exceeding 50%, measure the IC ₅₀ value of the compound on the tumor cells. In each experiment, two positive compounds of cisplatin (DDP) and paclitaxel (Taxol) were set, the concentration was taken as the abscissa, and the cell viability was plotted as the ordinate, and the cell growth curve was drawn, and the two-point method (Reed and Muench method) was used to calculate the compound _IC50 values.

According to the MTS method, with cisplatin (DDP) and paclitaxel (Taxol) as positive control compounds, a small amount of 7-(2,4-difluorophenyl)-1,3-dihydroxyxanthone synthesized in Example 1 was taken (Compound I for short) was dissolved in DMSO to make a 40 μM/L solution, and then the inhibitory activity of the above nine tumor cells was detected (Table 1-3).

Table 1 The inhibitory activity of compound Ⅰ to 5 kinds of tumor cells

Table 2 The inhibitory activity of compound Ⅰ to 4 kinds of tumor cells

Table 3 IC ₅₀ values of compound Ⅰ on three kinds of tumor cells

It can be seen from Table 1-3 that 7-(2,4-difluorophenyl)-1,3-dihydroxyxanthone has inhibitory effects on 9 kinds of tumor cells, among which human lung cancer A-549, human The inhibitory rate of liver cancer SMMC-7721 and human T-cell leukemia MT-4 cells exceeds 50%, and the IC ₅₀ value is between 18-25 μM, so 7-(2,4-difluorophenyl)-1,3-dihydroxy Xanthones have good antitumor activity against these three kinds of human tumor cells.

As can be seen from Table 3, the IC ₅₀ of 7-(2,4-difluorophenyl)-1,3-dihydroxyxanthone on lung cancer A-549 was 24.89±1.11 μM, while cisplatin on lung cancer A-549 The IC ₅₀ of -549 is 32.39±0.54 μM, so its inhibitory activity against lung cancer A-549 is stronger than that of cisplatin.

Example 3: Test of 7-(2,4-difluorophenyl)-1,3-dihydroxyxanthone inhibiting tyrosinase activity

Mix 7-(2,4-difluorophenyl)-1,3-dihydroxyxanthone with L-Dopa, add tyrosinase (final concentration 25U/mL) to start the reaction, set 3 repetitions Well, set a blank control without 6-trifluoromethyl-1,3-dihydroxyxanthone and a positive control of Kojic Acid at the same time, room temperature, 5min, measure the OD value with a microplate reader, and the detection wavelength is 490nm. The inhibition rate of tyrosinase activity was calculated.

Inhibition rate of tyrosinase activity (%) = (1 - sample OD _490nm / experimental control well OD _490nm ) × 100

The inhibitory effect of table 4 compound Ⅰ on tyrosinase

As can be seen from Table 4: 7-(2,4-difluorophenyl)-1,3-dihydroxyxanthone has the effect of inhibiting tyrosinase to a certain extent (inhibition rate is nearly 20%).

In summary, the compound 7-(2,4-difluorophenyl)-1,3-dihydroxyxanthone synthesized by the present invention is a new xanthone compound, and there is no literature or patent on it. Synthesis and biological activities are reported. The compound is effective against leukemia HL-60, T-cell leukemia MT-4, lung cancer A-549, liver cancer SMMC-7721, liver cancer HepG2, liver cancer Huh-7, colon cancer SW480, breast cancer MCF-7 and cervical cancer Hela cells Cells have inhibitory effect, among them, it has strong inhibitory effect on human lung cancer A-549, human liver cancer SMMC-7721 and human T-cell leukemia MT-4 cells, especially the inhibitory effect on lung cancer A-549 is better than clinical Cisplatin, which is widely used in the world, is also stronger and can be applied to the production of antitumor drugs.

At the same time, 7-(2,4-difluorophenyl)-1,3-dihydroxyxanthone also has the effect of inhibiting tyrosinase to a certain extent, so it has good transformation or application prospects, and can be widely used Drugs, health products, and foods used in the regulation of tyrosinase metabolism.

It should be noted that in this article, relational terms such as first and second are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply that there is a relationship between these entities or operations. any such actual relationship or order exists between them. Furthermore, the term "comprises", "comprises" or any other variation thereof is intended to cover a non-exclusive inclusion such that a process, method, article, or end-equipment comprising a set of elements includes not only those elements, but also includes elements not expressly listed. other elements identified, or also include elements inherent in such a process, method, article, or end-equipment. Without further limitations, an element defined by a statement "comprising..." or "comprising..." does not exclude the presence of additional elements in the process, method, article or terminal equipment comprising said element. In addition, in this article, "greater than", "less than", "exceeding", etc. are understood as not including the original number; "above", "below", "within" and so on are understood as including the original number.

Although the above-mentioned embodiments have been described, those skilled in the art can make additional changes and modifications to these embodiments once they know the basic creative concept, so the above-mentioned are only the implementation of the present invention For example, it is not intended to limit the scope of patent protection of the present invention. Any equivalent structure or equivalent process transformation made by using the content of the description of the present invention, or directly or indirectly used in other related technical fields, is also included in the patent of the present invention. within the scope of protection.

Claims (10)

1. one kind 2, the xanthone compound of 4- difluorophenyls substitution, it is characterised in that the compound is 7- (2,4- bis- Fluorophenyl) -1,3- dihydroxy xanthones, shown in its structure such as chemical formula (I)：

2. the preparation method of compound described in a kind of claim 1, it is characterised in that including by 5- (2,4- difluorophenyl) -2- The process that hydroxybenzoic acid, phloroglucin, phosphorus pentoxide and Loprazolam are reacted.

3. preparation method according to claim 2, it is characterised in that 5- (2,4- difluorophenyl) -2 hydroxybenzoic acids with The mol ratio of phloroglucin is 1:1.

4. preparation method according to claim 2, it is characterised in that comprise the following steps：

1) phosphorus pentoxide is dissolved in Loprazolam, prepares the first mixed solution；

2) the first mixed solution is added into 5- (2,4- difluorophenyl) -2 hydroxybenzoic acids and phloroglucin, carries out ring-closure reaction, Generate 7- (2,4 difluorobenzene base) -1,3- dihydroxy xanthones.

5. preparation method according to claim 2, it is characterised in that further comprising the steps of：

The solution containing 7- (2,4 difluorobenzene base) -1,3- dihydroxy xanthones after the completion of the step 2) is reacted is analysed Go out, filter, column chromatography for separation, 7- (2,4- difluorophenyl) -1, the 3- dihydroxy xanthones purified.

6. preparation method according to claim 4, it is characterised in that in the step 1), solution temperature is controlled 105 ~115 DEG C.

7. preparation method according to claim 4, it is characterised in that in the step 2), reaction temperature is controlled 88 ~92 DEG C, the reaction time is controlled in 18~22min.

8. the xanthone compound of the 2,4 difluorobenzene base substitution described in claim 1 is in terms of tyrosinase activity is suppressed Application.

9. xanthone compound the answering in terms of antineoplastic of the 2,4 difluorobenzene base substitution described in claim 1 With.

10. the xanthone compound of the 2,4 difluorobenzene base substitution described in claim 1 is in tyrosinase Metabolism regulation medicine Application in terms of thing, health products, food, cosmetics.