CN116589456A - AIE phototherapy agent with excellent I-type photodynamic activity and photothermal effect, and preparation method and application thereof - Google Patents

AIE phototherapy agent with excellent I-type photodynamic activity and photothermal effect, and preparation method and application thereof Download PDF

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CN116589456A
CN116589456A CN202310547941.6A CN202310547941A CN116589456A CN 116589456 A CN116589456 A CN 116589456A CN 202310547941 A CN202310547941 A CN 202310547941A CN 116589456 A CN116589456 A CN 116589456A
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aie
phototherapy agent
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杨晓静
汤显蛟
陈丽萍
赵美霞
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Henan University
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Abstract

The present invention relates to an AIE phototherapy agent having excellent type I photodynamic activity and photothermal effect, having the structure shown below:

Description

AIE phototherapy agent with excellent I-type photodynamic activity and photothermal effect, and preparation method and application thereof
Technical Field
The invention belongs to the technical field of organic synthesis and nano-drugs, and particularly relates to an AIE phototherapy agent with excellent I-type photodynamic activity and photothermal effect, a preparation method and application thereof in preparing anti-tumor drugs.
Background
At present, the traditional means for treating cancers, including techniques of surgical treatment, chemotherapy, radiotherapy and the like, have obvious limitations, such as toxic and side effects and unsatisfactory curative effects. Phototherapy is a non-invasive, efficient, low cost cancer treatment technique, including photodynamic therapy (PDT) and photothermal therapy (PTT).
PDT uses photosensitizing agents to intersystem crossing (ISC) to excited triplet states (T) under excitation by light 1 ），T 1 Highly cytotoxic reactive oxygen species ROS are produced by both type i (electron transfer) and type ii (energy transfer) mechanisms. Phototherapy has been attracting attention in recent years by using heat generated by light energy to kill cancer cells. The AIE-type photoactive molecules reported to date are based primarily on production 1 O 2 The oxygen concentration is severely dependent on the type II PDT, and the hypoxic environment of the tumor can seriously affect the PDT curative effect. In contrast, the type I photosensitizer has low oxygen dependence, and can kill cancer cells by generating active oxygen through disproportionation reaction, fenton reaction, etc. even under severe hypoxia condition. The type I photodynamic property of the photosensitive molecule is important to overcome the hypoxia problem faced in the photodynamic therapy of tumors and enhance the phototherapy effect. The AIE-type photoactive molecules reported to date are based primarily on production 1 O 2 How to design highly efficient type I photosensitizing molecules remains a challenging problem.
Disclosure of Invention
In view of this, the present invention provides an AIE phototherapy agent having excellent type I photodynamic activity and photothermal effect. The phototherapy agent has excellent I-type photodynamic activity and photothermal effect, and the nanoparticle prepared from the phototherapy agent can be used for passively targeting tumor tissues and can realize the combined treatment of PDT and PTT.
The invention also provides a preparation method of the AIE phototherapy agent with excellent I-type photodynamic activity and photothermal effect and application of the AIE phototherapy agent in preparation of antitumor drugs.
In order to achieve the above purpose, the invention adopts the following technical scheme:
an AIE phototherapy agent having excellent type I photodynamic activity and photothermal effect, having a structure represented by formula I:
；
formula I.
The AIE phototherapy agent with excellent type I photodynamic activity and photothermal effect comprises triphenylamine modified phenothiazine with AIE activity and bithiophene-linked malononitrile modified indane. The structure shown in the formula I generates I-type active oxygen under the excitation action of 660 and nm and has excellent photo-thermal conversion effect. The invention utilizes Bovine Serum Albumin (BSA) to load AIE phototherapy agent to prepare nano medicine/reagent, the phototherapy agent has hydrophobic inner core which can enter the BSA, the BSA has good biocompatibility, and the bioavailability of the phototherapy agent can be effectively improved.
The invention provides a preparation method of the AIE phototherapy agent, which comprises the following steps:
1) Mixing 10-ethylphenothiazine, N-bromosuccinimide (NBS) and a solvent I, and performing a first-step substitution reaction at room temperature for 10-15 h to obtain a compound PB;
2) Mixing a compound PB, 4-triphenylamine borate, tetra (triphenylphosphine) palladium, anhydrous sodium carbonate and a solvent II, and carrying out a second-step esterification reaction at 80-100 ℃ under the protection of nitrogen for 10-14 h to obtain a compound TPB;
3) Mixing a compound TPB,5 '-formyl-2, 2' -bithiophene-5-phenylboronic acid (CAS number: 1369328-62-0), tetrakis (triphenylphosphine) palladium, anhydrous sodium carbonate and a solvent three, and carrying out a third chemical reaction (esterification reaction) at 80-100 ℃ under the protection of nitrogen for 10-14 h to obtain a compound TPSS;
4) Mixing a compound TPSS, 1, 3-bis (dicyanomethylene) indane and a solvent IV, and carrying out a fourth chemical reaction at 55-70 ℃ for 10-15 h to obtain an AIE phototherapy agent, namely TPSSI with a structure shown in a formula I;
the structural formulas of the compounds PB, TPB and TPSS are shown in the following formulas II, III and IV respectively:
a formula II;
formula III;
formula IV.
Specifically, in the step 1), the molar ratio of the 10-ethylphenothiazine to the N-bromosuccinimide is preferably 1:2-2.5; the solvent used in the first substitution reaction is preferably N, N-Dimethylformamide (DMF). The temperature of the substitution reaction is preferably room temperature and the time is preferably 12 h.
Further, in the step 2), the molar ratio of the compound PB, the triphenylamine 4-borate, the tetrakis (triphenylphosphine) palladium and the anhydrous sodium carbonate is preferably 0.9-1:1:0.05-0.1:1.0-2, more preferably 1:1:0.05-0.08:1.3-1.8; the second solvent used in the second esterification reaction is preferably a mixture of water, toluene and absolute ethyl alcohol, and the volume ratio of the second solvent to the absolute ethyl alcohol is preferably 7-9:23-27:7-9, and more preferably 8:25:8. The temperature is preferably 80-100 ℃, more preferably 85-95 ℃, and the time of the esterification reaction is preferably 10-14 h, more preferably 12-13 h. In a specific embodiment of the present invention, PB, triphenylamine-4-borate, tetrakis (triphenylphosphine) palladium are preferably dissolved in toluene, then anhydrous sodium carbonate is dissolved in water, and then the solution is added to ethanol and stirred for the second chemical reaction. The purification method of TPB is not particularly limited in the present invention, and methods well known to those skilled in the art may be employed.
Specifically, in the step 3), the molar ratio of the compound TPB to the 5 '-formyl-2, 2' -bithiophene-5-phenylboronic acid to the tetra (triphenylphosphine) palladium to the anhydrous sodium carbonate is 1:1-1.5:0.5-0.8:1-2, and more preferably 1:1-1.2:0.6-0.7:1.1-1.3; the solvent III is preferably a mixture of water, toluene and absolute ethyl alcohol, and the volume ratio of the solvent III to the mixture is preferably 7-9:23-27:7-9, and more preferably 8:25:8. The temperature of the chemical reaction is preferably 80-100 ℃, more preferably 85-95 ℃ under the protection of nitrogen, and the time of the esterification reaction is preferably 10-14 h, more preferably 12-13 h. In a specific embodiment of the present invention, it is preferable to dissolve TPB, triphenylamine-4-borate, tetrakis (triphenylphosphine) palladium in toluene, then dissolve anhydrous sodium carbonate in water, and then add the solution to ethanol with stirring to perform the third chemical reaction.
Further, in the step 4), the molar ratio of the compound TPSS to the 1, 3-bis (dicyanomethylene) indane is preferably 1:1-1.5, more preferably 1:1-1.1; the fourth solvent is preferably acetic anhydride. The reaction temperature is preferably 50 to 80 ℃, more preferably 55 to 65 ℃.
In the above method for preparing AIE phototherapeutic agent, it is further preferable that after the fourth step of chemical reaction is completed, the excess acetic anhydride solvent is neutralized preferably with NaOH, dichloromethane is extracted, the crude product obtained by extraction is washed with ethanol and n-hexane respectively to remove unreacted raw materials, and then recrystallized with mixed solution of chloroform-n-hexane to obtain the product.
The invention also provides a nano reagent containing the AIE phototherapy agent, which comprises drug-loaded nano particles and a solvent, wherein the nano carrier is BSA, and the nano carrier BSA is activated by glutathione; the preparation method comprises the following steps: dissolving AIE phototherapy agent in tetrahydrofuran solvent to prepare 1-2 mg/mL TPSSI mother liquor; dissolving nano carrier BSA in PBS buffer solution, adding glutathione, ultrasound, adding TPSSI mother liquor, continuing ultrasound, removing tetrahydrofuran, then probe ultrasound, dialyzing to remove residual tetrahydrofuran and glutathione, and obtaining the nano carrier BSA.
In the invention, the solvent of the nano-carrier is preferably PBS buffer solution, and the pH value of the phosphate buffer solution is preferably 7.4; the solvent of the AIE phototherapeutic agent is preferably tetrahydrofuran. Specifically, the mass ratio of the nano-carrier BSA to the AIE phototherapy agent is preferably 20:0.5-5, and more preferably 20: 1-2. The particle size of the drug-carrying nano reagent is preferably 66-75 nm, more preferably 67.21 +/-0.32 nm.
In the present invention, the dialysis bag for dialysis preferably has a molecular weight cut-off of 2000Da, the dialysis time is preferably 24 hours, and water is preferably changed every 4 hours during the dialysis process; the nanoparticle form can effectively improve the bioavailability of the phototherapy agent.
In the present invention, the loading amount of TPSSI in the nano-agent containing the AIE phototherapy agent is preferably 4.25% ± 0.05% by mass; the calculation method of the load capacity is shown in a formula a:
w%=m 1 /m 2 x 100% formula a;
in formula a:w% is the loading, m 1 Is the mass, m of TPSSI contained in the nano-drug structure 2 The mass of the nano reagent.
The invention also provides application of the AIE phototherapy agent or the nano agent in preparation of antitumor drugs, and further preferably application in preparation of antitumor drugs.
The source of the 10-ethylphenothiazine is not particularly limited in the present invention, and can be either purchased directly from commercial sources or synthesized by methods well known to those skilled in the art.
The AIE phototherapy agent structure provided by the invention comprises triphenylamine modified phenothiazine AIE functional groups and bithiophene-linked malononitrile modified indane. The organic micromolecule has excellent I-type photodynamic activity and photothermal effect, and can realize the targeted delivery of chemotherapeutic drugs by using Bovine Serum Albumin (BSA) to load the organic micromolecule, thereby effectively improving the bioavailability of the phototherapy agent. The nanoparticle loaded phototherapy agent can realize the synergistic treatment of photodynamic therapy and photothermal therapy guided by fluorescence imaging, and improve the treatment efficiency of cancers, especially breast cancers.
Compared with the prior art, the invention has the following beneficial effects:
in the nano reagent provided by the invention, the phenothiazine has AIE activity, can promote fluorescence emission in an aggregation state, remarkably improve fluorescence intensity, can increase the imaging function on tumors, and realizes diagnosis and treatment integration; the nano reagent medicine is used for treating tumors, the nano medicine carries the phototherapy agent to be passively targeted and enriched in tumor tissues, then PDT and PTT are carried out for synergistic treatment, and the PDT-PTT combined treatment is used for killing tumor cells, especially breast cancer cells, so that the treatment efficiency is improved.
Drawings
FIG. 1 is a graph showing the variation of the temperature of the prepared TPSSI (100. Mu.M) with the laser irradiation time after 10 minutes of 660 nm laser irradiation;
FIG. 2 shows TPSSI or BSA-TPSSI NP S (TPSSI content 10. Mu.M), fluorescence intensity of Dichlorofluorescein (DCFH) at 525 nm was varied with 660 nm laser irradiation, I/I 0 -1 value change curve; the fluorescence intensity increases rapidly with the prolonged irradiation time;
FIG. 3 shows detection of type II using ABDA 1 O 2 The change in peak intensity of each maximum absorption peak of the ABDA+TPSSI (10. Mu.M) group compared to the ABDA (blank) group was negligible under 660 nm laser (0.3W) light irradiation, indicating the resulting form II 1 O 2 Can be ignored. The change of the absorption values of the ABDA at the wavelength of 381nm with time can be found that the ultraviolet absorption change of the experimental group is negligible compared with that of the blank group;
FIG. 4 is a graph of the validation of the generation of type I ROS by fluorescence probe, dihydrorhodamine DHR123 as an indicator. I/I of TPSSI (10. Mu.M) +DHR123 group 0 The value of-1 is greatly enhanced with the increase of the irradiation time of 660 nm laser (0.3W);
FIG. 5 is a nanoparticle BSA-TPSSI NP S Is a transmission electron microscope image;
FIG. 6 is a nanoparticle BSA-TPSSI NP S Temperature rise profile in aqueous solution (TPSSI content 100 μm) under laser irradiation at power 0.8. 0.8W;
FIG. 7 shows BSA-TPSSI NP S Effect on MCF-7 cell viability; in the figure, dark: no laser irradiation; light:660 Laser irradiation at nm (0.8W/cm) 2 ）；
FIG. 8 is a hydrogen spectrum of the AIE phototherapy agent (i.e., TPSSI) prepared in example 1.
Detailed Description
The following describes the technical scheme of the present invention in further detail with reference to examples, but the scope of the present invention is not limited thereto.
In the following examples, unless otherwise indicated, all the starting materials were either commercially available products which were commercially available as such or prepared according to methods conventional in the art. Room temperature refers to 25±5 ℃.
Example 1
An AIE phototherapy agent with excellent type I photodynamic activity and photothermal effect is prepared by the following steps:
1) NBS (2.39 g, 13.4 mmol) was added in portions to DMF (10 mL), immersed in ice water bath for cooling; and a solution of 10-ethylphenothiazine (1.49 g,6.56 mmol) in DMF (30 mL) was added dropwise and stirred overnight (12 h) at room temperature, the reaction mixture quenched with ice water. The organic product was extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered to remove solids, and concentrated by rotary evaporation. Purifying the crude product by column chromatography with n-hexane/ethyl acetate (15:1, volume ratio) as eluent to obtain white solid, namely compound PB;
2) Compound PB (384.90 mg,1 mmol), triphenylamine 4-borate (286.13 mg,1 mmol), tetrakis (triphenylphosphine) palladium (75 mg,0.064 mmol), anhydrous sodium carbonate (1.69 g,1.6 mmol) were added to a solvent (composed of a mixture of water 8 ml, toluene 25 ml and anhydrous ethanol 8 ml), and reacted under reflux at 90 ℃ under nitrogen protection overnight. After cooling, the reaction solution was poured into water, and the organic phase was separated by extraction with methylene chloride and dried. Passing the crude product through a silica gel column by taking DCM/PE (dichloromethane/petroleum ether=1:4, volume ratio) as an eluent to obtain a white solid product, namely a compound TPB;
3) Compound TPB (400 mg,0.72 mmol), 5 '-formyl-2, 2' -dithiophene-5-phenylboronic acid (259.99 mg,0.86 mmol), tetrakis (triphenylphosphine) palladium (54.16 mg,0.047 mmol) and anhydrous sodium carbonate (1.22 g,1.15 mmol) were added to a solvent (consisting of water 8 ml, toluene 25 ml and anhydrous ethanol 8 ml mixed) and refluxed overnight at 90 ℃ under nitrogen. After cooling, the reaction solution was poured into water, and the organic phase was separated by extraction with methylene chloride and dried. Passing the crude product through a silica gel column by taking DCM/PE (1:2, volume ratio) as an eluent to obtain a yellow pasty product, namely a compound TPSS;
4) Compound TPSS (150 mg,0.226 mmol) was dissolved in acetic anhydride (10 mL), 1, 3-bis (dicyanomethylene) indane (54.74 mg,0.226 mmol) was added to the solution, and the resulting mixture was stirred at 60℃overnight. After cooling, acetic anhydride was neutralized with 0.1M NaOH, followed by extraction with dichloromethane and drying of the organic phase, the resulting solid product was washed with ethanol to remove unreacted indane and with n-hexane to remove unreacted compound TPSS. Recrystallizing the obtained crude product by using a chloroform/normal hexane mixed solution (1:10, volume ratio) to obtain a blue-black solid product, namely TPSSI;
the structural formulas of the compounds PB, TPB and TPSS are shown in the following formulas II, III and IV respectively:
a formula II;
formula III;
formula IV.
The AIE phototherapy agent (namely TPSSI) with excellent type I photodynamic activity and photothermal effect has the following structural formula:
。
the hydrogen spectrum data of AIE phototherapy (i.e. TPSSI) is shown in FIG. 8, TPSSI 1 H NMR (300 MHz, DMSO-d 6 ) δ 9.88 (s, 1H), 8.00 (d, J = 4.1 Hz, 1H), 7.95 – 7.88 (m, 2H), 7.62 – 7.57 (m, 2H), 7.54 (d, J = 7.1 Hz, 3H), 7.49 (s, 2H), 7.43 (d, J = 9.0 Hz, 4H), 7.32(t, J = 7.7 Hz, 4H), 7.04 (q, J = 10.0, 9.4 Hz, 9H), 5.73 (d, J = 14.8 Hz, 2H), 3.97 (d, J = 7.0 Hz, 2H), 1.34 (t, J = 6.9 Hz, 3H)。
Example 2
A nanoagent comprising the AIE phototherapeutic agent prepared by the steps of:
TPSSI 1.5. 1.5 mg is weighed and dissolved in 1.1 mL tetrahydrofuran to prepare 1.5. 1.5 mg/mL of TPSSI mother liquor for later use. 30 mg nanometer BSA carrier is dissolved in PBS buffer solution (pH 7.4) of 10 mL, glutathione of 6 mg is added, and ultrasound is carried out10 minutes. The TPSSI mother liquor of 1 ml was added and the ultrasound continued for 20 minutes. Removing tetrahydrofuran by rotary evaporation under reduced pressure, performing ultrasonic treatment with a probe for 10 min, and dialyzing for 24h (Mw=2000 Da, i.e. dialysis bag with molecular weight cut-off of 2000Da, and changing water every 4h during dialysis) to remove residual tetrahydrofuran and glutathione to obtain nanometer reagent BSA-TPSSI NP S Freeze-drying (-80 ℃, 24-h), and preserving at 4 ℃ for later use.
The obtained BSA-TPSSI NP S The morphology of the nano particles is shown in a transmission electron microscope of FIG. 5; as can be seen from FIG. 5, BSA-TPSSI NP S The nanoparticle size was about 70 a nm a.
Example 3
A DMSO solution of TPSSI having a TPSSI content of 100. Mu.M was irradiated with 660 nm laser light at 0.8W/cm 2 The change in solution temperature with time was recorded over 10 minutes, and the temperature was measured every 30 seconds, with the results shown in fig. 1.
As can be seen from fig. 1: along with the extension of the laser irradiation time, the temperature of the TPSSI solution is gradually increased to 69.5 ℃ which is far higher than the temperature of DMSO, which indicates that the TPSSI provided by the invention has excellent photo-thermal effect and can carry out photo-thermal treatment.
An aqueous solution of BSA-TPSSI NPs having a TPSSI content of 100. Mu.M was irradiated with 660 nm laser light at 0.8W/cm 2 The change in solution temperature with time was recorded over 10 minutes, and the temperature was measured every 30 seconds, with the results shown in fig. 6.
As can be seen from fig. 6: along with the extension of the laser irradiation time, the temperature of the TPSSI NPs solution is gradually increased to 57.5 ℃ and is far higher than the temperature of water, which indicates that the TPSSINPs provided by the invention have excellent photo-thermal effect and can be used for photo-thermal treatment.
Example 4
Ethanol solution of DCFH-DA (reactive oxygen species fluorescent probe) (1X 10) -3 M, 0.5. 0.5 mL) was added to 2 mLNaOH (1X 10) -2 M) and stirred at room temperature for 30 minutes, DCFH-DA was hydrolyzed to DCFH. Then, 10 mL of PBS buffer solution (pH 7.4) was added to the mixture to stop the hydrolysis and stored in the dark until use. 0.5ml of 100. Mu.M TPSSI or BSA-TPSS was takenAqueous mother liquor of I NPs, DCFH 1.25 ml (40 μm) was added and the PBS buffer solution was diluted to 5ml to obtain a solution of TPSSI or BSA-TPSSI NPs with a TPSSI content of 10 μm. 660 Laser irradiation at nm (0.3W/cm) 2 ) Photoluminescence spectra under 488 nm excitation were measured at different time intervals and emissions were collected in the range 490 to 600 nm. With the maximum photoluminescence intensity per irradiation (I) and the initial photoluminescence intensity without irradiation (I 0 ) Calculation of active oxygen production efficiency (I/I) 0 -1), the results are shown in FIG. 2.
As can be seen from fig. 2: along with the extension of irradiation time, the fluorescence intensity of the TPSSI+DCFH-DA group and the BSA-TPSSI NPs+DCFH-DA group at 525 nm is obviously increased compared with that of the DCFH-DA group, which proves that the TPSSI phototherapy agent and the BSA-TPSSI NPs provided by the invention have excellent photodynamic effect and can carry out photodynamic therapy.
Example 5
The singlet oxygen generating capacity of TPSSI under 660 nm laser irradiation was evaluated using 9, 10-anthracenediyl-bis (methylene) bis-malonic acid (ABDA). The prepared buffer solution of ABDA (50. Mu.M) and TPSSI (5. Mu.M) in PBS (5 ml) was exposed to 660 nm laser irradiation (0.3W/cm) 2 ). Absorbance of ABDA at 378 nm was recorded at different irradiation times, singlet oxygen was detected, and the results are shown in fig. 3.
As can be seen from fig. 3: with the extension of the laser irradiation time, the absorbance of the ABDA+TPSSI group at 378 nm is negligible compared with the absorbance of the ABDA group, which indicates that the TPSSI provided by the invention does not generate type II active oxygen.
Example 6
The dihydrorhodamine 123 (DHR 123) is used for free radical detection, 3.463 mg dihydrorhodamine 123 is taken and diluted to 10 ml, and 1 mmol/L DHR123 mother liquor is obtained. TPSSI solids 2.78, mg were taken and diluted to 2.5, mL to give a 1 mmol/L TPSSI mother liquor. mu.L of TPSSI stock solution and 50. Mu.L of DHR123 stock solution were taken and diluted to 5mL with PBS buffer. Irradiation with 660 nm laser (0.3W/cm) 2 ) Photoluminescence spectra under 488 and nm excitation are measured at different time intervals, and emission in the range of 490-600 nm is collected. Maximum with each irradiationPhotoluminescence intensity (I) and initial photoluminescence intensity without irradiation (I 0 ) Calculation of active oxygen production efficiency (I/I) 0 -1), the results are shown in FIG. 4.
From fig. 4 and as can be seen: along with the extension of irradiation time, compared with the DHR123 group, the fluorescence intensity of the TPSSI+DHR123 group is obviously increased, which proves that the TPSSI phototherapy agent provided by the invention can generate I-type active oxygen O 2 −• Type I photodynamic therapy is enabled.
Example 7
Cytotoxicity of BSA-TPSSI NPs was detected by MTT assay. Inoculating MCF-7 cells of human breast cancer cells in logarithmic growth phase into 96-well plate, wherein the number of cells per well is about 7×10 3 ~8×10 3 Putting into 37 ℃ and 5 percent CO 2 Is cultured in a cell culture incubator of 24 h. BSA-TPSSI NP with different concentrations S (0.5, 2, 5, 10, 20. Mu.M/L) on MCF-7 cells. After culturing 12h, the cells were irradiated with 660 nm laser (0.8W/cm) 2 ) After 10 min per well, it was returned to the incubator for further incubation for 12 h. 50 mu L of MTT is added into each hole, 4h is continuously cultivated in an incubator, 100 mu L of DMSO is added into each hole, shaking table is vibrated for 10 min, and absorbance is measured by an enzyme-labeled instrument when the purple crystals are completely dissolved. Calculation of nano-drug BSA-TPSSI NP S The effect on cell viability is shown in FIG. 7.
Cell viability (%) = [ a570 (dosing) -a570 (blank) ]/[ a570 (negative) -a570 (blank) ]x100% formula B;
as can be seen from the results in fig. 7: BSA-TPSSI NP S The survival rate of the MCF-7 cells after being treated is obviously reduced compared with that of the cells before being irradiated by laser, and the effect is optimal especially when the concentration is 10 and 20 mu M/L. Description of BSA-TPSSI NP S The PTT and PDT combined treatment of the (E) can obviously inhibit the growth of tumors.

Claims (10)

1. An AIE phototherapy agent having excellent type I photodynamic activity and photothermal effect, characterized by having a structure represented by the following formula I:
；
formula I.
2. The method for preparing AIE phototherapy agent according to claim 1, comprising the steps of:
1) Mixing 10-ethyl phenothiazine, N-bromosuccinimide and a solvent I, and performing a first-step substitution reaction at room temperature for 10-15 h to obtain a compound PB;
2) Mixing a compound PB, 4-triphenylamine borate, tetra (triphenylphosphine) palladium, anhydrous sodium carbonate and a solvent II, and carrying out a second-step esterification reaction at 80-100 ℃ under the protection of nitrogen for 10-14 h to obtain a compound TPB;
3) Mixing a compound TPB,5 '-formyl-2, 2' -bithiophene-5-phenylboric acid, tetra (triphenylphosphine) palladium, anhydrous sodium carbonate and a solvent three, and carrying out a third chemical reaction at 80-100 ℃ under the protection of nitrogen for 10-14 h to obtain a compound TPSS;
4) Mixing a compound TPSS, 1, 3-bis (dicyanomethylene) indane and a solvent IV, and performing a fourth chemical reaction at 50-70 ℃ for 10-15 hours to obtain the compound;
the structural formulas of the compounds PB, TPB and TPSS are shown in the following formulas II, III and IV respectively:
a formula II;
formula III;
formula IV.
3. The method of preparing AIE phototherapeutic agent according to claim 2, wherein in step 1), the molar ratio of 10-ethylphenothiazine and N-bromosuccinimide is 1:2-2.5; the first solvent is N, N-dimethylformamide.
4. The method for preparing an AIE phototherapy agent according to claim 2, wherein in step 2), the molar ratio of the compound PB, triphenylamine 4-borate, tetrakis (triphenylphosphine) palladium and anhydrous sodium carbonate is 0.9-1:1:0.05-0.1:1.0-2; the second solvent is a mixture of water, toluene and absolute ethyl alcohol.
5. The method for preparing an AIE phototherapy agent according to claim 2, wherein in step 3), the molar ratio of the compound TPB,5 '-formyl-2, 2' -bithiophene-5-phenylboronic acid, tetrakis (triphenylphosphine) palladium, anhydrous sodium carbonate is 1:1 to 1.5:0.5 to 0.8:1 to 2; and the solvent III is a mixture of water, toluene and absolute ethyl alcohol.
6. The method for preparing an AIE phototherapy agent according to claim 2, wherein in step 4), the molar ratio of the compound TPSS to 1, 3-bis (dicyanomethylene) indane is 1:1-1.5; and the solvent IV is acetic anhydride.
7. The method for preparing AIE phototherapy agent according to claim 6, wherein after the fourth step of chemical reaction, neutralizing the excessive acetic anhydride solvent with NaOH, extracting with dichloromethane, washing the crude product obtained by extraction with ethanol and n-hexane, and recrystallizing with mixed solution of chloroform and n-hexane.
8. A nanoreagent comprising the AIE phototherapy agent of claim 1, prepared by the steps of: dissolving AIE phototherapy agent in tetrahydrofuran to prepare 1-2 mg/mL TPSSI mother liquor; dissolving nano-carrier BSA in PBS buffer solution, adding glutathione, ultrasound, adding TPSSI mother liquor, continuing ultrasound, removing tetrahydrofuran, then ultrasound, dialyzing to remove residual tetrahydrofuran and glutathione, and obtaining the nano-carrier BSA.
9. The nano-agent according to claim 8, wherein the mass ratio of the nano-carrier BSA to the AIE phototherapy agent is 20:0.5-5, and the loading amount of the AIE phototherapy agent is 4.2-4.3%.
10. Use of an AIE phototherapy agent according to claim 1 or a nanoagent according to claim 8 for the preparation of an antitumor drug.
CN202310547941.6A 2023-05-16 2023-05-16 AIE phototherapy agent with excellent I-type photodynamic activity and photothermal effect, and preparation method and application thereof Pending CN116589456A (en)

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