CN111205669A - Method for synthesizing anthracene fluorescent dye - Google Patents

Abstract

The invention provides a method for synthesizing anthracene fluorescent dye, which solves the problems of difficult preparation and low yield of key intermediate in the synthesis of anthracene fluorescent dye, and simultaneously has the advantages of easily obtained raw materials, mild reaction conditions and reduced operation danger. The synthetic method mainly comprises six steps, m-halogenated phenol or substituted m-halogenated phenol reacts with formaldehyde in an acid solution, and the diphenylmethane derivative is obtained through purification; namely a first intermediate product; converting the phenolic hydroxyl of the first intermediate product into methyl ether or silicon-based ether to obtain a diphenylmethane derivative with a phenolic hydroxyl protecting group; a second intermediate product; converting the second intermediate product into an organic lithium salt and reacting with a dihalo-compound to produce an anthracene derivative, i.e. a third intermediate product; oxidizing the third intermediate product to obtain an anthrone compound, namely a fourth intermediate product; reacting the fourth intermediate product with a grignard reagent, a fifth intermediate product; and removing the phenolic hydroxyl protecting group to obtain the fluorescent dye.

Description

Method for synthesizing anthracene fluorescent dye

Technical Field

The invention relates to a synthesis method of near-infrared band anthracene fluorescent dye, belonging to the field of biochemical synthesis. Is especially suitable for synthesizing anthracene compounds and anthracene compounds containing hetero atoms.

Background

With the rapid development of high-throughput sequencing technology in recent years, compared with the traditional sanger sequencing, the method has the greatest advantage that massive sequence information can be read simultaneously. Although the accuracy is not as good as that of the traditional sequencing method, the information beyond the sequence itself can be obtained due to the analysis of mass data, such as gene expression amount, copy number variation and more accurate reading, which is an important direction for the development of high-throughput sequencing.

Fluorogenic sequencing methods are also widely used in high-throughput sequencing, which uses terminal phosphate-labeled nucleotide substrates. Fluorogenic substrates are of the general formula: one class of molecules is modified at the end or in the middle of the 5' polyphosphate with a fluorophore with fluorescence switching properties:

characterized by a fluorogenic fluorophore modified at the terminal phosphate of deoxyribonucleotides (including A, C, G, T, U and others) of 4, 5, 6, or more phosphates, and no label at the base and 3' hydroxyl group. This fluorophore when modified on the phosphate is different from the absorption spectrum and/or emission spectrum of the phosphate when it is detached.

The anthracene fluorescent dye is used as a fluorescein analogue, namely carbon-bridge substituted fluorescein or silicon-bridge substituted fluorescein, has the characteristics of long wavelength, fluorescence producibility, simple and easy derivatization, high molar absorption coefficient, high quantum yield and the like, and can be used for marking nucleotide and applied to fluorogenic high-throughput sequencing technology. Among them, one of the labeling techniques is to connect different fluorescent groups to specific positions of nucleotide bases through a cleavable long chain, and remove the labeled groups by an additional chemical means after the DNA completes recognition and records a fluorescent signal, thereby preparing for the next round of nucleotide introduction. The applicant's prior patent CN201510155218.9 also discloses anthracene fluorescent dyes and a method for synthesizing the same. However, the synthesis yield is low, and the reaction conditions are relatively limited. The invention adopts a novel method for synthesizing anthracene fluorescent dye to solve the problems that key intermediate is difficult to prepare and the yield is low during synthesizing anthracene fluorescent dye; meanwhile, the raw materials required by the new synthetic route are cheap and easy to obtain, the reaction conditions are milder, the risk of reaction operation is reduced, and the harm to the environment is less in the experimental process and the experimental treatment.

Disclosure of Invention

The purpose of the invention is realized by the following technical scheme. The invention provides a method for synthesizing anthracene dye with fluorescence switching property, which is characterized by comprising the following steps,

1) m-halogenated phenol or substituted m-halogenated phenol reacts with formaldehyde in an acid solution, and the diphenylmethane derivative is obtained through purification; namely a first intermediate product;

2) converting the phenolic hydroxyl of the first intermediate product into methyl ether or silicon-based ether to obtain a diphenylmethane derivative with a phenolic hydroxyl protecting group; a second intermediate product;

3) converting the second intermediate product into an organic lithium salt and reacting with a dihalo-compound to produce an anthracene derivative, i.e. a third intermediate product;

4) oxidizing the third intermediate product to obtain an anthrone compound, namely a fourth intermediate product;

5) reacting the fourth intermediate product with a grignard reagent, a fifth intermediate product;

6) and removing the phenolic hydroxyl protecting group to obtain the fluorescent dye.

According to a preferred embodiment, the substituted meta-halophenol is a halogenated phenol having a substituent in at least one of the 2,6 positions.

According to a preferred embodiment, the substituted meta-halophenol refers to C1-C6 alkyl, halogenated C1-C6 alkyl.

According to a preferred embodiment, in step (1), the m-halophenol or substituted m-halophenol is reacted with formaldehyde in an acidic solution, meaning that the m-halophenol or substituted m-halophenol is reacted with paraformaldehyde in an acidic heated solution,

according to a preferred embodiment, in the step (1), the heating reaction time is 10-24h, and the heating temperature is 40-50 ℃.

According to a preferred embodiment, in step (5), the Grignard reagent is selected from the group consisting of a phenylmagnesium chloride Grignard reagent, or a phenyllithium Grignard reagent.

According to a preferred embodiment, in step (3), the dihalo-compound is a diphenyl or dialkyl-substituted dihalomethane, or a diphenyl or dialkyl-substituted dihalosilane.

The invention discloses a method for synthesizing anthracene dye with fluorescence switching property, which is characterized by comprising the following steps,

1) the phenolic hydroxyl group of the m-halophenol or the substituted m-halophenol is converted into methyl ether or silicon ether,

namely a first intermediate product;

2) reacting the first intermediate product with formaldehyde in an acid solution, and purifying to obtain a diphenylmethane derivative; i.e. the second intermediate product;

3) converting the second intermediate product into an organic lithium salt and reacting with a dihalo-compound to produce an anthracene derivative, i.e. a third intermediate product;

4) oxidizing the third intermediate product to obtain an anthrone compound, namely a fourth intermediate product;

5) reacting the fourth intermediate product with a grignard reagent, a fifth intermediate product;

6) and removing the phenolic hydroxyl protecting group to obtain the fluorescent dye.

The invention discloses a synthetic method of anthracene fluorescent dye, which has the following advantages: (1) the raw materials of the new route are cheap and easy to obtain. (2) The intermediate is simple to prepare and high in yield. (3) Avoiding the use of organic metal compounds, and being more environment-friendly. (4) The operation and synthesis are simple and easy.

Detailed Description

Exemplary embodiments of the present disclosure are described in more detail below. It should be understood that the present disclosure may be embodied in various forms and should not be limited by the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the disclosure to those skilled in the art.

The applicant has disclosed a series of anthracene compounds in previous patents, see patent CN 201510155218.9. If necessary, part of the disclosure in this patent may be incorporated by reference into this patent.

Except where expressly specified, all terms in this description are to be understood as commonly understood in the art.

The anthracene compound referred to in the present invention means a compound having a structure similar to triphenylanthracene. This term is not intended to be a special term.

Similar terms such as C1-C6 are common terms in the chemical field. C1-C6 means containing 1-6 carbon atoms.

In the reaction related to the present invention, other products exist in some steps, but the target product can be easily obtained by those skilled in the art on the basis of the description of the present invention. The specific structure of the by-product is not the core of the present invention and is not described too much.

The invention discloses a method for synthesizing anthracene compounds, which is characterized by comprising the following steps of (1) reacting m-halogenated phenol or substituted m-halogenated phenol with formaldehyde in an acid solution, and purifying to obtain diphenylmethane derivatives; namely a first intermediate product; (2) converting the phenolic hydroxyl of the first intermediate product into methyl ether or silicon-based ether to obtain a diphenylmethane derivative with a phenolic hydroxyl protecting group; a second intermediate product; (3) converting the second intermediate product into an organic lithium salt and reacting with a dihalo-compound to produce an anthracene derivative, i.e. a third intermediate product; (4) oxidizing the third intermediate product to obtain an anthrone compound, namely a fourth intermediate product; (5) reacting the fourth intermediate product with a Grignard reagent, a fifth intermediate product; (6) and removing the phenolic hydroxyl protecting group to obtain the fluorescent dye.

Generally, phenol is more reactive at the para position. The m-halophenol used in the present invention means that at least one of the 2,6 positions of the halophenol has a substituent. The substituent can be common substituent with nonreactive property, such as methyl, ethyl, C1-C6 alkyl, halogen substituted C1-C6 alkyl and the like.

Phenolic polycondensation is a common chemical reaction. In the phenol molecule, a P orbit where lone pair electrons of oxygen atoms in phenolic hydroxyl are located and a pi electron orbit of a large pi bond conjugated by a benzene ring are mutually crossed and covered to form a conjugated system, so that a P electron cloud on the oxygen atoms is transferred to the benzene ring, the electron cloud density on the benzene ring is increased, the benzene ring is activated, the electron cloud density of the hydroxyl at the ortho-position and the para-position is increased, the activity of hydrogen atoms is enhanced, in the formaldehyde molecule, the electron cloud density on the carbon atoms is reduced due to the attraction of the oxygen atoms, and the hydrogen atoms at the ortho-position or the para-position of the phenol and the carbonyl of the formaldehyde are subjected to addition reaction under the action of acid or alkali to generate the hydroxymethyl phenol. Under the catalysis of acid (commonly used concentrated hydrochloric acid), because the acid catalyst inhibits the ionization of phenol, the conjugation of lone pair electrons of hydroxyl oxygen atoms of phenol and benzene rings is reduced, the electron cloud density of the benzene rings is increased little, the catalytic action of the acid enhances the reaction activity of formaldehyde and hydroxymethyl phenol, when the phenol is excessive, the phenol mainly reacts at two ortho positions to generate hydroxymethyl phenol, and the hydroxymethyl phenol and the phenol are mutually dehydrated and condensed to form the linear phenolic resin. When the reactants and conditions are controlled, a polycondensation product of two molecules can be obtained. See, for example, the first reaction step in scheme 1 below.

The present invention will be further described with reference to specific route examples. See scheme 1 below:

(1) m-halogenated phenol or substituted m-halogenated phenol reacts with formaldehyde solution under the acidic heating condition to obtain an addition product, namely a diphenylmethane derivative, wherein the position of a substituent can be 2 position, 6 position or 2 position and 6 position and simultaneously has a substituent; or m-halogenated anisole or substituted m-halogenated anisole reacts with formaldehyde under acidic conditions to directly obtain the diphenylmethane derivative with the protecting group.

(2) Converting the hydroxyl of the diphenylmethane derivative into methyl ether or silicon-based ether for protection (such as adding sodium hydride to remove hydrogen on the hydroxyl and reacting with methyl iodide to generate halogenated anisole, and reacting phenolic hydroxyl with chlorosilane compounds under the catalysis of imidazole to generate silicon ether), thereby obtaining the diphenylmethane derivative with a hydroxyl protecting group.

(3) The brominated diphenylmethane derivatives are firstly converted into organic lithium salts, and then react with dihalogenated compounds to form anthracene derivatives.

(4) Oxidizing the anthracene derivative into the anthrone compound by an oxidant.

(5) The anthracene ketone compound with the protecting group reacts with the Grignard reagent to obtain the aryl-substituted anthracene derivative.

(6) Removing phenolic hydroxyl protecting groups to finally obtain different types of fluorescent compounds.

Can be explained. As can be appreciated by those skilled in the art through this step. The protection step of the phenolic hydroxyl group is not fixed. For example, the phenolic hydroxyl group may be first methylated or silylized protected and then reacted.

It will be appreciated that in step 5, the format reagent may generally be adapted to be a newly formed format reagent. May be phenylmagnesium chloride Grignard reagent, phenyllithium, etc.; the phenyl group may be a substituted phenyl group.

It will be appreciated that in step 3, both halogens are removed as the dihalo-compound is reacted. The compound distorts to form a new six-membered ring.

More specifically, the following will explain the second route in more detail as an example. The principle of the two routes and the one route are completely the same. However, specific compounds are used for some compounds in the process. Scheme 2 below:

except for the description of scheme 2. The invention is also exchangeable in other ways. For example, scheme 3 below:

1 the synthetic routes listed herein are general formulas of the synthetic routes developed this time, and the substitutable functional groups are all replaced with R.

2 scheme 2 herein is a scheme for the synthesis of carbon heteroanthracene fluorescent dyes and scheme 3 is a scheme for the synthesis of silanthracene fluorescent dyes. Scheme 2 differs from scheme 3 in that different anthracene-based fluorescent dyes (e.g., 2-dibromopropane and dichlorodimethylsilane used in the third step) were synthesized using different halogenating agents.

And 3, summarizing: the method described in scheme 1 is clearly different from the prior method for synthesizing the heteroanthracene compound (CN105315698B) by the applicant. The method comprises the following steps: (1) the key point of the synthesis method is to synthesize the heteroanthrone derivative firstly and synthesize the fluorescent dye precursor by utilizing the reaction of the heteroanthrone derivative and the Grignard reagent. The key step of the previous method is to synthesize the fluorescent dye precursor by a methanesulfonic acid heated cyclization method. (2) The prior synthesis method uses toxic and harmful reagents such as organic zinc, titanium tetrachloride and the like, and the feeding is not easy to operate, and the method adopts conventional reagents. (3) The method obviously improves the total yield because of optimizing the key step of low ring closure yield of ketone and benzene ring under the acidic condition. The overall yield of the present invention was 40% and the overall yield of the previous invention was approximately 10%. 4 abbreviations for the compounds used herein are as follows: TBSCl is tert-butyldimethylsilyl chloride; THF, tetrahydrofuran; DMF is N, N-dimethylformamide; DMSC is dichlorodimethylsilane; TBAF tetrabutylammonium fluoride.

It can be illustrated that in scheme 2, dimethyldibromomethane (2, 2-dibromopropane) is used during the synthesis of compound 3-4; as can be seen from the analysis of the specific synthetic route. In the latter reaction, two methyl groups remain in the anthracene compound. Therefore, the fluorescent material can be replaced by various other compounds without affecting the fluorescence performance of the whole compound. The compound is characterized in that two hydrogens in a methane molecule are replaced by halogens, and the other two hydrogens are replaced by other structures. And various alternatives to the other structures described herein are possible. For example, methyl, ethyl, propyl, isopropyl, substituted or unsubstituted C1-C6 alkyl. Halogen substituted C1-C6 alkyl. And so on. These substitutions do not result in too much change of the compound. Meanwhile, it is to be noted that the two substituents may be the same or different.

The invention discloses a method for synthesizing anthracene compounds, which is characterized by comprising the following steps:

(1) taking m-halogenated phenol or substituted m-halogenated phenol; dissolving in methanol water solution of concentrated hydrochloric acid; adding paraformaldehyde, heating to 40-60 ℃, and reacting for 10-40 hours; separating the product to obtain a diphenylmethane derivative; namely a first intermediate product;

(2) reacting the first intermediate product with tert-butyldimethylsilyl chloride to obtain a diphenylmethane derivative with a phenolic hydroxyl protecting group TBS; a second intermediate product;

(3) reacting the second intermediate product with n-butyllithium to obtain an organic lithium salt, and reacting the organic lithium salt with a dihalogenated compound 2, 2-dibromopropane to obtain an anthracene derivative, namely a third intermediate product;

(4) oxidizing the third intermediate product by potassium permanganate to obtain an anthrone compound, namely a fourth intermediate product;

(5) reacting the fourth intermediate product with a Grignard reagent isopropyl magnesium chloride lithium chloride, a fifth intermediate product;

(6) and removing the phenolic hydroxyl protecting group to obtain the fluorescent dye.

In the following examples, all anhydrous operations were carried out under nitrogen and the reaction vessels used were dried. The chemical raw materials, reagents, solvents and the like used are, unless otherwise specified, indicated as analytical pure reagents which are commercially available as they are. The anhydrous treatment of the used dry solvent is carried out according to a conventional method or the anhydrous super-dry solvent is directly purchased.

Method and apparatus for the isolation and identification of products: silica gel of thin layer chromatography column is GF 25460 (Merk), 254nm and 365nm double-wavelength portable ultraviolet detector; the column chromatography silica gel is 200-plus 300-mesh silica gel of Qingdaozhiyida silica gel reagent limited company; measuring the nuclear magnetic resonance spectrum by using a VArian300 nuclear magnetic resonance instrument; hydrogen spectrum, carbon spectrum.

Example 1

The synthesis of the fluorescent dye COOH-PO is shown in a synthetic route 2;

1) synthesis of compound 4,4' -methyenebis (3-bromopenol) 2: compound 1(10g, 57.8mmol) was weighed out, dissolved in 40% concentrated HCl in methanol and water (1:1), and paraformaldehyde (360mg, 4mmol) was added to heat the reaction mixture to 50 ℃ for 24 hours. Most of methanol in the reaction solution was removed by rotary column chromatography, and then 23.6 g of an oily compound was obtained by adding brine, extracting with dichloromethane, and purifying by rotary column chromatography.

1H NMR(300MHz,CDCl3)δ8.95(s,2H),7.22(dt,J＝7.5,1.1Hz, 2H),7.01(d,J＝1.5Hz,2H),6.75(dd,J＝7.5,1.5Hz,2H),4.02(t, J＝1.0Hz,2H).13C NMR(75MHz,CDCl3)δ156.32,131.75,127.53,125.48, 118.98,118.58,42.17.HRMS:Caled for C13H11Br2O2(M+H),358.0290, Found m/z 358.0291。

2) Synthesis of the compound bis (2-bromoo-4- ((tert-butylmethylistylyl) oxy) phenyl) methane 3: dissolving the compound 2(4g,11.3mmol) in 20ml DMF, adding tert-butyldimethylsilyl chloride (5g,33.8mmol), adding imidazole (4.6g, 67.8mmol), reacting at room temperature for 8 hours, monitoring the reaction by TCL until the reaction is completed, adding water for washing, extracting by ethyl acetate, removing the solvent by rotation, and separating by column chromatography to obtain 36 g of oily compound.

1H NMR(300MHz,CDCl3)δ7.23–6.96(m,4H),6.78(dd,J＝7.5, 1.5Hz,2H),4.03(t,J＝1.0Hz,2H),1.01(s,18H),0.21(s,12H).13C NMR(75MHz,CDCl3)δ150.52,135.85,128.72,116.45,116.01,109.27, 35.30,25.93,18.36,-3.53.HRMS:Caled forC25H39Br2O2Si2(M+H), 586.5550,Found m/z 586.5548。

3) Synthesis of compound ((9,9-dimethyl-9, 10-dihydrothracene-2, 7-dimethyl) bis (oxy)) bis (tert-butyl dimethyl) 4: dissolving a compound 3(1.5g, 2.56mmol) in 5ml of anhydrous THF, cooling to-78 ℃ under the protection of argon, slowly dropping n-butyllithium (4.1ml, 5.3mmol) for reaction for 1 hour, adding 2, 2-dibromopropane (0.3ml, 2.8mmol) and slowly raising the temperature to room temperature for reaction for 24 hours, adding water for quenching reaction, adding a saturated aqueous solution of sodium chloride, extracting with ethyl acetate, and removing a solvent for purification to obtain 41.3 g of a white solid compound.

1H NMR(300MHz,CDCl3)δ7.08(dt,J＝7.5,1.0Hz,2H),6.87 (dd,J＝11.7,1.5Hz,2H),6.73(dd,J＝7.5,1.5Hz,2H),3.81(t,J ＝1.0Hz,2H),1.57(s,6H),1.01(s,18H),0.21(s,12H).

13C NMR(75MHz,CDCl3)δ151.63,143.72,130.02,129.98,127.35, 119.77,114.81,114.79,40.05,35.12,31.08,25.54,17.56,-4.57.HRMS: Caled for C28H45O2Si2(M+H),468.2880,Found m/z 468.2876

4) Synthesis of the compound 3,6-bis ((tert-butylmethylisolyl) oxy) -10, 10-dimethyllantracen-9 (10H) -one 5: compound 4(0.58g, 1.0mmol) was dissolved in 5ml of acetone solution, potassium permanganate (1.0g, 7mmol) was added under 1 ice bath to react at 0 ℃ or lower for 3 hours, 10ml of dichloromethane was added, and insoluble matter was filtered off to give 50.25 g of a white solid compound.

1H NMR(300MHz,CDCl3)δ7.88(d,J＝7.5Hz,2H),7.02(dd,J＝ 7.5,1.6Hz,2H),6.85(d,J＝1.6Hz,2H),1.59(s,9H),1.01(s,24H), 0.21(s,17H)..13C NMR(75MHz,CDCl3)δ182.48,156.25,145.73,129.32, 128.95,118.83,114.76,40.75,30.96,25.54,17.56,-4.57.HRMS:Caled for C28H43O3Si2(M+H),482.8110,Found m/z 482.8108

5) The compound methyl 4- (3,6-bis (tert-butylmethylilyl) oxy) -9-hydroxy-10,10-dimethyl-9, 10-

dihydroanthracenone-9-yl) -3-me-tylbenzoate 7 synthesis: dissolving compound 6(0.8mg, 2.8mmol) in 5ml of anhydrous tetrahydrofuran solution, adding 2.9mmol of isopropyl magnesium chloride lithium chloride, and reacting at room temperature for 2 hours; the tetrahydrofuran solution containing compound 5(1.3mg, 2.8mmol) was slowly added to the freshly prepared Grignard reagent and reacted at room temperature for 8 hours; TCL detects the reaction condition and adds 5ml of water after the reaction is completed, and the stirring reaction is continued for 5 hours; the solvent was removed by rotation, and the red solid compound 7200 mg was obtained by column chromatography with ethyl acetate.

1H NMR(300MHz,CDCl3)δ8.11–7.84(m,1H),7.79(dd,J＝7.5, 1.5Hz,1H),7.23(d,J＝7.5Hz,1H),7.17(d,J＝7.5Hz,2H),6.96 –6.66(m,4H),3.93(d,J＝14.3Hz,4H),2.35(s,3H),1.57(s,6H), 1.02(s,18H),0.21(s,12H).13C NMR(75MHz,CDCl3)δ166.40,152.55, 143.72,142.11,135.48,132.55,132.25,130.53,129.01,128.68,127.21,126.85,119.91,116.20,74.75,52.11,40.04,30.96,25.54,21.90,17.54, -4.57.HRMS:Caled for C37H53O5Si2(M+H),632.9880,Found m/z 632.9879

6) The compound 4- (6-hydroxy-10,10-dimethyl-3-oxo-3,10-dihydroanthracen-9-yl) -3-meth ylbenzoic

acid 8 synthesis: dissolving 70.2 g of the compound in 1ml of methanol solution, adding 1ml of concentrated hydrochloric acid, 2ml of water and 0.5g of sodium fluoride, and continuously stirring for reacting for 5 hours; and (4) removing the solvent by spinning. Dissolving the crude product in 5ml mixed solution of methanol and water 1:1, adding lithium hydroxide (75mg, 3mmol), reacting at room temperature for 10 hr, hydrolyzing completely, removing methanol, extracting with saturated saline solution ethyl acetate, and purifying by column chromatography to obtain 850 mg red solid compound.

1H NMR(300MHz,CDCl3)δ8.74(s,1H),7.97–7.77(m,2H),7.50 –7.29(m,3H),6.91(d,J＝1.4Hz,1H),6.77(dd,J＝7.5,1.5Hz, 1H),6.68–6.46(m,2H),2.43(s,3H),1.56(s,6H).13C NMR(75MHz, CDCl3)δ185.60,168.99,156.26,151.59,145.24,141.46,139.53,137.64, 136.58,132.42,130.53,130.47,129.93,128.61,128.17,128.10,127.26,121.32,115.68,113.27,38.88,30.40,20.16.

HRMS:Caled for C24H22O4(M+H),372.4200,Found m/z 372.4210

Detailed description of the preferred embodiment

Synthesis of the fluorescent dyes of COOH-TM is shown in scheme 3

1 Synthesis of Compound 4,4' -Methylenbis (3-bromopenol) 2:

compound 1(10g, 57.8mmol) was weighed out, dissolved in 40% concentrated HCl in methanol and water (1:1), and reacted at room temperature for 20 minutes, and paraformaldehyde (360mg, 4mmol) was added to heat the reaction mixture to 50 ℃ for 24 hours. Most of methanol in the reaction solution was removed by rotary column chromatography, and then 23.6 g of an oily compound was obtained by adding brine, extracting with dichloromethane, and purifying by rotary column chromatography.

1H NMR(300MHz,CDCl3)δ8.95(s,2H),7.22(dt,J＝7.5,1.1Hz, 2H),7.01(d,J＝1.5Hz,2H),6.75(dd,J＝7.5,1.5Hz,2H),4.02(t, J＝1.0Hz,2H).13C NMR(75MHz,CDCl3)δ156.32,131.75,127.53,125.48, 118.98,118.58,42.17.HRMS:Caled for C13H11Br2O2(M+H),358.0290, Found m/z 358.0291。

2 Synthesis of Compound bis (2-bromoo-4- ((tert-butylmethylistylyl) oxy) phenyl) methane 3:

dissolving the compound 2(4g,11.3mmol) in 20ml DMF, adding tert-butyldimethylsilyl chloride (5g,33.8mmol), adding imidazole (4.6g, 67.8mmol), reacting at room temperature for 8 hours, monitoring the reaction by TCL until the reaction is completed, adding water for washing, extracting by ethyl acetate, removing the solvent by rotation, and separating by column chromatography to obtain 36 g of oily compound.

1H NMR(300MHz,CDCl3)δ7.23–6.96(m,4H),6.78(dd,J＝7.5, 1.5Hz,2H),4.03(t,J＝1.0Hz,2H),1.01(s,18H),0.21(s,12H).13C NMR(75MHz,CDCl3)δ150.52,135.85,128.72,116.45,116.01,109.27, 35.30,25.93,18.36,-3.53.HRMS:Caled forC25H39Br2O2Si2(M+H), 586.5550,Found m/z 586.5548。

3 Synthesis of the Compound 3,7-bis (tert-butyldithiolyl) oxy) -5,5-dimethyl-5,10-dihydrodiben zo [ b, e ] sine 9: dissolving the compound 3(8.5g, 14.5mmol) in 50ml of anhydrous THF, cooling to-78 ℃ under the protection of argon, slowly dropping n-butyllithium (12.7ml, 30.6mmol) for reaction for 1 hour, adding dichlorodimethylsilane (2ml, 16mmol) and slowly raising to room temperature for reaction for 24 hours, adding water for quenching reaction, adding a saturated aqueous solution of sodium chloride, extracting with ethyl acetate, and removing the solvent by spinning to purify to obtain 4.8g of the compound.

1H NMR(300MHz,CDCl3)δ7.24(dt,J＝7.5,1.0Hz,2H),7.02(t, J＝1.7Hz,2H),6.84(dd,J＝7.5,1.5Hz,2H),4.33(t,J＝1.0Hz, 2H),1.01(s,18H),0.46(s,6H),0.21(s,12H).13C NMR(75MHz,CDCl3) δ153.05,137.65,134.17,127.65,122.64,121.92,37.73,25.54,17.55, -3.15,-4.57.HRMS:Caled for C27H45O2Si3(M+H),484.9020,Found m/z484.9019

Synthesis of 4 Compound 3,7-bis ((tert-butylmethylisolyl) oxy) -5, 5-dimethylibenzo [ b, e ] silin-10 (5H) -one 10: compound 9(4.8g, 10.3mmol) was dissolved in 15ml of acetone solution, potassium permanganate (4.4g, 27.8mmol) was added under ice bath to react at 0 ℃ or lower for 3 hours, 100ml of dichloromethane was added, and insoluble matter was filtered off to give 103.5 g of a red solid compound.

1H NMR(300MHz,CDCl3)δ8.00(d,J＝7.5Hz,2H),7.19–7.11 (m,4H),1.01(s,18H),0.50(s,6H),0.21(s,12H).13C NMR(75MHz,CDCl3) δ184.28,156.43,137.38,135.80,135.49,128.49,122.34,120.47,25.54, 17.56,-3.25,-4.57.HRMS:Caled forC27H43O3Si3(M+H),498.8850,Found m/z 498.8847.

5 Compound methyl 4- (7-hydroxy-5,5-dimethyl-3-oxo-3, 5-dihydrodibenzo)

[ b, e ] silin-10-yl) -Methylbenzoate11 Synthesis: dissolving compound 6(1.3g, 4.6mmol) in anhydrous tetrahydrofuran solution, adding 4.6mmol of isopropyl magnesium chloride lithium chloride, and reacting at room temperature for 2 hours; slowly adding a tetrahydrofuran solution dissolved with the compound 10 into a newly prepared Grignard reagent, and reacting at room temperature for 8 hours; TCL detects the reaction condition, 5ml of concentrated hydrochloric acid, 20ml of water and 1ml of tetrabutyl ammonium fluoride are added after the reaction is completed, and the mixture is continuously stirred and reacts for 5 hours; removing solvent, and purifying with ethyl acetate column chromatography to obtain red solid compound 11500 mg.

1H NMR(300MHz,CDCl3)δ9.11(s,1H),8.09–8.02(m,2H),7.87 –7.83(m,1H),7.62(d,J＝7.5Hz,1H),7.48(d,J＝7.5Hz,1H),6.97 (d,J＝1.4Hz,1H),6.85(dd,J＝7.5,1.5Hz,1H),6.80(d,J＝2.2 Hz,1H),6.68(dd,J＝10.8,2.2Hz,1H),3.92(s,3H),2.47(s,3H), 0.53(s,6H).13C NMR(75MHz,CDCl3)δ185.53,166.48,157.84,146.96, 143.49,142.48,139.51,137.59,137.03,135.42,132.15,130.07,129.97, 129.86,129.67,129.37,127.47,125.03,118.60,118.07,52.09,20.16, -3.46.HRMS:Caled forC24H23O4Si(M+H),402.5210,Found m/z 402.5212

Synthesis of 4- (7-hydroxy-5,5-dimethyl-3-oxo-3,5-dihydrodibenzo [ b, e ] silin-10-yl) -3-methyllbenzoic acid 12, a compound of 6: dissolving compound 11(500mg, 1.243mmol) in a mixed solution of methanol and water 1:1, adding lithium hydroxide (522mg, 12.4mmol) to react at room temperature for 10 hours, after complete hydrolysis, removing methanol by rotation, adding saturated saline water, extracting with ethyl acetate, and purifying by column chromatography to obtain 12300 mg of red solid compound.

1H NMR(300MHz,CDCl3)δ9.11(s,1H),8.05(d,J＝11.0Hz,1H), 7.91–7.86(m,2H),7.62(d,J＝7.5Hz,1H),7.48(d,J＝7.7Hz,1H), 6.97(d,J＝1.5Hz,1H),6.85(dd,J＝7.5,1.5Hz,1H),6.80(d,J＝ 2.2Hz,1H),6.68(dd,J＝10.8,2.2Hz,1H),2.47(s,3H),0.53(s,6H).13C NMR(75MHz,CDCl3)δ185.53,168.99,157.84,146.96,145.22,142.48,139.51,137.86,136.91,135.46,132.14,130.43,129.97,129.37,129.20, 129.06,128.10,125.03,118.60,118.08,20.13,-3.47.HRMS:Caled for C23H21O4Si(M+H),388.4940,Found m/z 388.4939

The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are included in the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the appended claims.

Claims (8)

1. A method for synthesizing anthracene fluorescent dye is characterized by comprising the following steps,

(1) m-halogenated phenol or substituted m-halogenated phenol reacts with formaldehyde in an acid solution, and the diphenylmethane derivative is obtained through purification; namely a first intermediate product;

(2) converting the phenolic hydroxyl of the first intermediate product into methyl ether or silicon-based ether to obtain a diphenylmethane derivative with a phenolic hydroxyl protecting group; a second intermediate product;

(3) converting the second intermediate product into an organic lithium salt and reacting with a dihalo-compound to produce an anthracene derivative, i.e. a third intermediate product;

(4) oxidizing the third intermediate product to obtain an anthrone compound, namely a fourth intermediate product;

(5) reacting the fourth intermediate product with a Grignard reagent to obtain a fifth intermediate product;

(6) and removing the phenolic hydroxyl protecting group to obtain the fluorescent dye.

2. The method of claim 1, wherein said substituted meta-halophenol is substituted at least in one of the 2,6 positions of the halophenol.

3. The method of claims 1-2, wherein the substituted meta-halophenol is C1-C6 alkyl, halogenated C1-C6 alkyl.

4. The method according to any one of the preceding claims, wherein in step (1), the m-halophenol or substituted m-halophenol is reacted with formaldehyde in an acidic solution, which means that the m-halophenol or substituted m-halophenol is reacted with paraformaldehyde in an acidic heated solution.

5. The method according to any one of the preceding claims, wherein in step (1), the reaction is heated for 10-24h at a temperature of 40-50 ℃.

6. The method of any preceding claim, wherein in step (5) the Grignard reagent is selected from phenylmagnesium chloride Grignard reagents, or phenyllithium, or substituted phenylmagnesium chloride Grignard reagents, or substituted phenyllithium Grignard reagents.

7. The process according to any one of the preceding claims, characterized in that in step (3) the dihalo-compound is a diphenyl or dialkyl substituted dihalomethane or a diphenyl or dialkyl substituted dihalosilane.

8. A method for synthesizing anthracene fluorescent dye is characterized by comprising the following steps,

(1) converting the phenolic hydroxyl group of the m-halophenol or the substituted m-halophenol into methyl ether or silicon ether, namely a first intermediate product;

(2) reacting the first intermediate product with formaldehyde in an acid solution, and purifying to obtain a diphenylmethane derivative; i.e. the second intermediate product;

(3) converting the second intermediate product into an organic lithium salt and reacting with a dihalo-compound to produce an anthracene derivative, i.e. a third intermediate product;

(4) oxidizing the third intermediate product to obtain an anthrone compound, namely a fourth intermediate product;

(5) reacting the fourth intermediate product with a Grignard reagent to obtain a fifth intermediate product;

(6) and removing the phenolic hydroxyl protecting group from the fifth intermediate product to obtain the fluorescent dye.