CN107759803B

CN107759803B - Tetrasulfhydryl porphyrin compound and preparation method and application thereof

Info

Publication number: CN107759803B
Application number: CN201711114753.5A
Authority: CN
Inventors: 董育斌; 姜嫄; 阚京兰
Original assignee: Shandong Normal University
Current assignee: Shandong Normal University
Priority date: 2017-11-13
Filing date: 2017-11-13
Publication date: 2020-07-03
Anticipated expiration: 2037-11-13
Also published as: CN107759803A

Abstract

The invention relates to the field of functional complexes, in particular to a tetra-sulfhydryl porphyrin compound and a preparation method and application thereof; the tetra-sulfhydryl porphyrin prepared by the method has strong coordination capacity, and is combined with a UiO-66 framework material to prepare a tetra-sulfhydryl porphyrin modified metal organic framework material (UiO-66), namely UiO-66-H₂TPP‑(SH)₄The porphyrin derivative has very good water solubility, well solves the problems of difficult dissolution with water, easy agglomeration and poor stability of porphyrin molecular compounds in the prior art, and greatly improves the quantum yield, the fluorescence life and the photosensitive property of singlet oxygen of the porphyrin and the porphyrin derivative.

Description

Tetrasulfhydryl porphyrin compound and preparation method and application thereof

Technical Field

The invention relates to the field of functional complexes, and particularly relates to a tetrakismercaptoporphyrin compound, and a preparation method and application thereof.

Background

Porphyrins and their derivatives are a typical class of planar pi-conjugated macrocyclic molecules, which are widely found in nature and in living beings. The porphyrin ring is used as the core part of the compound and is composed of a closed and continuous conjugated porphin ring formed by connecting four pyrrole rings through methine (-CH ═) respectively. The periphery of the porphyrin ring can be substituted by various functional groups to form various substituted porphyrins. Porphyrins occurring in nature are usually substituted by other functional groups on the ring of conjugated porphines, whereas artificially synthesized porphyrins are substituted mainly by four methines, i.e. meso-positions (meso positions). Because the meso-site (meso-site) modified porphyrin compound has high synthesis yield and good stability, researchers often use the substituted porphyrin compound as a research object. Research shows that porphyrin compounds have unique physiological activity and affinity with cancer cells, and thus are often used as anticancer photosensitizers for photodynamic therapy (PDT) in the medical field. However, due to the structural characteristics of porphyrin and derivatives thereof, namely a large-plane pi conjugated system, the macrocyclic porphyrin compound has extremely low solubility in aqueous solution and exists in an aggregate form, so that the quantum yield and the fluorescence lifetime of singlet oxygen are greatly reduced, and the photosensitive property of the macrocyclic porphyrin compound is reduced.

In summary, the problems of poor solubility in water, easy agglomeration, poor stability, etc. still exist in the prior porphyrins and derivatives thereof, which seriously affect the chemical and optical properties of the porphyrins and derivatives thereof, and therefore, a tetrakismercaptoporphyrin compound and a preparation method thereof, which can overcome the problems, are needed.

Disclosure of Invention

Aiming at the problems in the prior art, the invention aims to provide a tetra-sulfhydryl porphyrin compound, a preparation method and application thereof, the tetra-sulfhydryl porphyrin prepared by the method has strong coordination capacity, and the tetra-sulfhydryl porphyrin compound is combined with a UiO-66 framework material to prepare a tetra-sulfhydryl porphyrin modified metal organic framework material (UiO-66-H), namely UiO-66-H₂TPP-(SH)₄The porphyrin derivative has very good water solubility, well solves the problems of difficult dissolution with water, easy agglomeration and poor stability of porphyrin molecular compounds in the prior art, and greatly improves the quantum yield, the fluorescence life and the photosensitive property of singlet oxygen of the porphyrin and the porphyrin derivative.

One of the objects of the present invention is to provide a tetrakismercaptoporphyrin.

The other purpose of the invention is to provide a preparation method of the tetrakismercaptoporphyrin.

The invention also aims to provide a preparation method of the metal organic framework material (UiO-66).

The fourth purpose of the invention is to provide a method for preparing UiO-66-H by modifying UiO-66 with tetrathioporphyrin₂TPP-(SH)₄The method of (1).

The fifth object of the present invention is to provide the above-mentioned UiO-66-H₂TPP-(SH)₄The use of (1).

In order to achieve the above purpose, the invention specifically discloses the following technical scheme:

firstly, the invention discloses a tetra-sulfhydryl porphyrin compound, and the structural formula of the tetra-sulfhydryl porphyrin compound is as follows:

the molecular formula of the tetra-sulfhydryl porphyrin compound is as follows: h₂TPP-(COSC₂H₄SH)₄5,10,15, 20-tetrakis (4- (S-2-mercaptoethyl) thiocarbonylphenyl) porphyrinTetrahydropulfhydryl porphyrin (H)₂TPP-(SH)₄) In which H is₂TPP is tetraphenylporphyrin.

Secondly, the invention discloses a preparation method of tetrasulfhydryl porphyrin, which specifically comprises the following steps:

(1) h is to be₂TPP-(COOH)₄Dissolving in an organic solvent, adding thionyl chloride for reaction and reflux, evaporating the solvent after the reflux is finished, and adding dried tetrahydrofuran to obtain a first reaction solution;

(2) dissolving 1, 2-dimercaptoethane and triethylamine in tetrahydrofuran to obtain a second reaction solution;

(3) dropwise adding the second reaction solution into the first reaction solution, and stirring and reacting for 1.5-3 h at room temperature;

(4) carrying out post-treatment on the solution obtained in the step (3) to obtain H₂TPP-(SH)₄。

In the step (1), the hydrogen atom₂TPP-(COOH)₄The mass-volume ratio (mg/mL) of the organic solvent to the thionyl chloride is (15-20): 1: 1;

preferably, the organic solvent is dichloromethane, chloroform, toluene, or the like.

In the step (1), the reaction conditions are as follows: the temperature is 40-45 ℃, and the reflux time is 4-5 h.

In the step (1), the hydrogen atom₂TPP-(COOH)₄The mass-to-volume ratio (mg/mL) of tetrahydrofuran to tetrahydrofuran is (5-7): 1.

in the step (2), the volume ratio of the 1, 2-dimercaptoethane to triethylamine is (10-20): 1.

in the step (2), the volume ratio of the 1, 2-dimercaptoethane to tetrahydrofuran is (0.02-0.06): 1.

step (1) H₂TPP-(COOH)₄And the mass ratio of the 1, 2-dimercaptoethane in the step (2) is 1: (60-80).

In the step (3), the dripping speed is 2-3 seconds per drop, and the dripping process is carried out at normal temperature and in a dark place.

In the step (4), the post-treatment comprises the following steps: performing column chromatography and recrystallization.

Preferably, the column chromatography adopts silica gel column chromatography, and dichloromethane is used as eluent; the recrystallization is carried out in a solvent of dichloromethane and methanol, and the volume ratio of the dichloromethane to the methanol is 1: (100-120).

The invention further discloses a preparation method of the metal organic framework material UiO-66, which comprises the following steps:

(1) reacting ZrCl₄Adding the mixture into DMF (N, N-dimethylformamide), adding acetic acid, finally adding terephthalic acid, ultrasonically dispersing uniformly, synthesizing a UiO-66 nano material by a solvothermal method, and performing high-speed centrifugal separation to obtain a solid product A;

(2) washing the solid product A in the step (1) with DMF to obtain a solid product B, and activating the solid product B to obtain white powder UiO-66.

In the step (1), ZrCl₄Mass ratio to terephthalic acid was 1: 1.

in the step (1), ZrCl₄The mass-to-volume ratio (mg/mL) of DMF to DMF is (4-6): 1.

in the step (1), the volume ratio of acetic acid to DMF is (0.035-0.075): 1.

in the step (1), the conditions for synthesizing the UiO-66 nano material by the solvothermal method are as follows: the temperature is 90-120 ℃, the heat preservation time is 12-48 h, and preferably, the heat preservation time is 24 h.

In the step (2), the activation is carried out by immersing ① the solid product A in a DMF solvent, stirring and immersing for 6-10 h at 60-85 ℃, carrying out high-speed centrifugal separation at ② to obtain a solid product B, immersing ③ the solid product B in absolute ethyl alcohol, stirring and immersing for 7-12 h at 50-75 ℃, carrying out high-speed centrifugal separation to obtain a solid product C, repeating the step ③ for 3 times to obtain a solid product D, immersing and washing the solid product D with ethyl ether, and airing under natural conditions to obtain a solid product E, thus obtaining the metal organic framework material UiO-66.

Finally, the tetrakismercaptoporphyrin and UiO-66 prepared by the invention are respectively used as a photosensitizer and a metal organic framework material, and the tetrakismercaptoporphyrin is used for modifying the metal organic framework material UiO-66 to obtain the metal organic framework material UiO-66Tetramercaptoporphyrin-modified UiO-66, namely UiO-66-H₂TPP-(SH)₄The mass ratio of the UiO-66 to the tetrasulfhydryl porphyrin is 11-13. Specifically, the tetrathioporphyrin is adopted to modify UiO-66 to prepare UiO-66-H₂TPP-(SH)₄The method comprises the following steps:

(a) adding the tetrathiol porphyrin into an organic solvent for dissolving to prepare a tetrathiol porphyrin solution;

(b) adding UiO-66 into the tetrathioporphyrin solution in the step (a), performing ultrasonic dispersion, and stirring;

(c) centrifuging the mixture obtained in the step (b) to obtain a solid product, washing the solid product with an organic solvent, activating the solid product, and drying to obtain the tetrakismercaptoporphyrin modified UiO-66, namely UiO-66-H₂TPP-(SH)₄。

In step (a), the organic solvent is dry dichloromethane.

In the step (a), the concentration of the substance in the tetrakismercaptoporphyrin solution is 0.5-3 mmol.L^-1。

In the step (b), the mass-to-volume ratio (mg/mL) of the UiO-66 to the tetrakismercaptoporphyrin solution is (2.5-5.0): 1.

in the step (b), the ultrasonic dispersion time is 20-25 min.

In the step (c), the centrifugal speed is 12000-13000 r/m.

In the step (c), the activation refers to immersing the solid product into dichloromethane, and stirring for 5-7 hours while soaking at normal temperature.

In addition, the invention also provides the UiO-66-H prepared by the invention₂TPP-(SH)₄Including use in photodynamic therapy (PDT), in particular as an anti-cancer photosensitizer.

The invention adopts UiO-66 to overcome the problems of difficult water solubility, easy agglomeration and poor stability of porphyrin molecules in the prior art, and the principle is as follows: the artificially synthesized porphyrin molecule is tetraphenyl substituted porphyrin, the porphyrin compound is rigid conjugated macromolecule, is easy to dissolve in organic solvent and insoluble in water, and is often in the form of aggregate in solutionThe formula (I) exists; if the porphyrin molecules are combined with other substances which can stably exist in the aqueous solution in a monodispersed manner, the problems that the porphyrin molecules are difficult to dissolve in water and easy to gather can be solved; the invention synthesizes dimercaptoethane group porphyrin compound H with strong coordination capacity₂TPP-(SH)₄The method combines the metal organic framework nano material with high stability and low biological toxicity in aqueous solution UiO-66, which utilizes the surface space defect of UiO-66 and the strong coordination capacity of dimercaptoethane group to synthesize H by post-modification₂TPP-(SH)₄Modifying the surface of the UiO-66 to obtain the nano material UiO-66-H₂TPP-(SH)₄(ii) a Thus, the Tetramercaptoporphyrin H of the present invention was used₂TPP-(SH)₄Post-modified synthetic UiO-66-H₂TPP-(SH)₄Not only maintains the internal framework structure of UiO-66, and has good dispersibility and stability in aqueous solution, but also maintains the photosensitive property of porphyrin, thereby being used as a novel anticancer photosensitive material in PDT.

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

(1) according to test results, the tetra-mercapto porphyrin compound obtained by the invention is an excellent photosensitizer, and the porphyrin molecule contains four dimercaptoethane groups with strong coordination capacity.

(2) According to the test result, the nano material UiO-66-H is obtained by modifying the tetra-sulfhydryl porphyrin compound obtained by the invention on the surface of UiO-66₂TPP-(SH)₄The material not only keeps the good dispersibility and stability of UiO-66 in aqueous solution, but also keeps the photosensitive characteristic of porphyrin, overcomes the problems of difficult water solubility, easy agglomeration, poor stability and the like of porphyrin molecules, and is an excellent anticancer photosensitive nano material.

Drawings

The accompanying drawings, which are incorporated in and constitute a part of this application, illustrate embodiments of the application and, together with the description, serve to explain the application and are not intended to limit the application.

FIG. 1 is the tetrasulfhydryl porphyrin H synthesized in example 1₂TPP-(SH)₄High resolution mass spectrograms of (1).

FIG. 2 is the tetrasulfhydryl porphyrin H synthesized in example 1₂TPP-(SH)₄Nuclear magnetic spectrum of (1).

FIG. 3 is the tetrasulfhydryl porphyrin H synthesized in example 1₂TPP-(SH)₄An infrared spectrum of (1).

FIG. 4 is the tetrasulfhydryl porphyrin H synthesized in example 1₂TPP-(SH)₄Ultraviolet absorption spectrum of (2).

FIG. 5 shows UiO-66 and UiO-66-H in example 4₂TPP-(SH)₄Scanning Electron Microscope (SEM) images of (a).

FIG. 6 shows UiO-66-H in example 4₂TPP-(SH)₄Transmission Electron Microscope (TEM) images of (a).

FIG. 7 shows UiO-66 and UiO-66-H in example 4₂TPP-(SH)₄Powder diffraction Pattern (PXRD).

FIG. 8 shows UiO-66-H in example 4₂TPP-(SH)₄Powder diffraction Patterns (PXRD) before and after soaking in phosphate buffer.

FIG. 9 shows UiO-66 and UiO-66-H in example 4₂TPP-(SH)₄Thermogravimetric analysis (TGA) of (a).

FIG. 10 shows UiO-66 and UiO-66-H in example 4₂TPP-(SH)₄Adsorption-desorption curve of (1).

FIG. 11 shows UiO-66 and UiO-66-H in example 4₂TPP-(SH)₄Distribution plot of pore size of (a).

FIG. 12 shows UiO-66 and H in example 4₂TPP-(SH)₄And UiO-66-H₂TPP-(SH)₄Ultraviolet absorption spectrum (UV-vis).

FIG. 13 shows the 415nm peak change of 1, 3-Diphenylisobenzofuran (DPBF) in example 7 under 660nm illumination.

FIG. 14 is the tetramercaptoporphyrin H of example 7₂TPP-(SH)₄The generation of singlet oxygen.

FIG. 15 shows UiO-66-H in example 7₂TPP-(SH)₄The generation of singlet oxygen.

Detailed Description

It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.

It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of example embodiments according to the present application. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof, unless the context clearly indicates otherwise.

It should be noted that H used in the present invention is for better technical effects₂TPP-(COOH)₄For self-preparation, but now commercially available H₂TPP-(COOH)₄The finished product is also suitable for use in the present invention.

As introduced in the background art, the existing porphyrins and derivatives thereof still have the problems of poor solubility in water, easy agglomeration, poor stability and the like, and in order to solve the problems, the invention provides a tetramercaptoporphyrin compound and a preparation method and application thereof, and the invention is further explained by combining specific examples.

Example 1

Tetramercaptoporphyrin H₂TPP-(SH)₄The preparation of (1):

(1)H₂TPP-(COOH)₄the preparation of (1):

methyl p-formylbenzoate (6.9g, 0.042mol) and 100mL of propionic acid were heated to 145 ℃ for reaction and refluxing to obtain a mixed solution, and then 20mL of propionic acid and pyrrole (3.0mL, 0.043mol) were added dropwise to the above mixed solution, and after the addition was completed, refluxing was continued for 1 h. Then, the mixture was naturally cooled to room temperature, and methanol was added thereto and allowed to stand for 12 hours. Finally, filtering, washing the filter cake with water and methanol to obtain purple powder H₂TPP-(COOCH₃)₄Its mass was 1.8g, the amount of substance was 0.0021mol, the yield was 20％。

Will be 1g H₂TPP-(COOCH₃)₄80mL of Tetrahydrofuran (THF) and 6mL of 10 mol. L^-1KOH and 100mL water are heated to 70 ℃ and refluxed for 6h, and then cooled to room temperature naturally. THF was removed by distillation under normal pressure and filtered. Then adjusting the pH of the filtrate to 1-2 with hydrochloric acid, carrying out suction filtration, washing the filter cake with water and methanol, and airing to obtain solid H₂TPP-(COOH)₄(ii) a The specific reaction process is as follows:

(2)H₂TPP-(SH)₄the preparation of (1):

h is to be₂TPP-(COOH)₄(79mg, 0.10mmol) was dissolved in 5mL of dry dichloromethane, and SOCl was added in the absence of light₂(5mL, 0.069mol) was slowly added dropwise to the mixed solution, and after refluxing at 40 ℃ for 5 hours, excess SOCl was distilled off under normal pressure₂And dichloromethane to obtain the tetra-acyl chloride phenyl porphyrin H in black green solid state₂TPP-(COCl)₄. 84mg of H₂TPP-(COCl)₄Dissolved in 15mL of dried THF to give a first reaction solution.

1, 2-dimercaptoethane (672. mu.L, 8.0mmol) and triethylamine (40. mu.L, 0.29mmol) were dissolved in 15mL of dry THF to obtain a second reaction solution.

Dropwise adding the first reaction solution into the second reaction solution at the normal temperature in a dark place at the dropping speed of 2 seconds/drop, and stirring for reaction for 3 hours after the dropwise adding is finished; then, evaporating the solvent by rotary evaporation, taking dichloromethane as an eluent, and separating and purifying the target product by silica gel column chromatography.

And finally, recrystallizing in dichloromethane and methanol, wherein the volume ratio of the dichloromethane to the methanol is 1: 100 to obtain a purple solid product H₂TPP-(SH)₄The mass was 29mg, the amount of substance was 0.026mmol, and the yield was 26%. The reaction process is as follows:

for H prepared in this example₂TPP-(SH)₄The results of the mass spectrometry, nuclear magnetic, infrared spectroscopy and ultraviolet spectroscopy are shown in fig. 1 to 4. Wherein, FIG. 1 is H₂TPP-(SH)₄The mass spectrum of the flight time high resolution, MALDI-TOF/MS, m/z calculation: c₅₆H₄₆N₄O₄S₈[M+H]⁺1095.1, corresponding to the peak at 1095.5 in fig. 1.

FIG. 2 is H₂TCCPP-(SH)₄The nuclear magnetic hydrogen spectrum of (a) is,¹H-NMR(300MHz，CDCl₃)ppm：δ＝8.82(s，8H，β-pyrrole)，8.37-8.40(d，8H，phenyl)，8.30-8.33(d，8H，phenyl)，3.46-3.51(m，8H，-CH₂CH₂-)，2.92-3.11(m，8H，-CH₂CH₂-)，1.77-1.83(t，4H，-SH)，-2.82(s，2H，NH)。

FIG. 3 is H₂TPP-(SH)₄IR spectrum of (a), IR: 3311(w), 2917(w), 2545(w), 1655(vs), 1597(m), 1472(vw), 1396(w), 1276(vw), 1204(s), 1184(m), 966(w), 920(w), 810(m), 709(w), 654 (vw).

H₂TPP-(SH)₄The ultraviolet absorption spectrum of (A) was measured in DMF, see FIG. 4, [ lambda ]_max(nm)(log(ε)，M^-1cm^-1)]:420(5.02)，516(3.72)，551(3.51)，591(3.31)，646(3.15)。H₂TPP-(SH)₄The compound has a strong porphyrin Soret band absorption at 420nm, and has Q band absorption of four typical free porphyrins at 516, 551, 591 and 646 nm.

As can be seen from FIGS. 1 to 4, the final product obtained in this example is Compound H₂TPP-(SH)₄。

Example 2

Tetramercaptoporphyrin H₂TPP-(SH)₄The preparation of (1):

(1)H₂TPP-(COOH)₄the preparation method of (1) is the same as that of example 1.

(2)H₂TPP-(SH)₄The preparation of (1):

h is to be₂TPP-(COOH)₄(90mg, 0.11mmol) was dissolved in6mL of dry dichloromethane, and then adding SOCl in the absence of light₂(6mL, 0.069mol) was slowly added dropwise to the mixed solution, reacted at 45 ℃ under reflux for 5 hours, and excess SOCl was evaporated off under normal pressure₂And dichloromethane to obtain the tetra-acyl chloride phenyl porphyrin H in black green solid state₂TPP-(COCl)₄. 93mg of H₂TPP-(COCl)₄Dissolved in 15mL of dried THF to give a first reaction solution.

1, 2-dimercaptoethane (672. mu.L, 8.0mmol) and triethylamine (40. mu.L, 0.29mmol) were dissolved in 25mL of dry THF to obtain a second reaction solution.

And dropwise adding the first reaction solution into the second reaction solution at normal temperature in a dark condition, wherein the dropwise adding speed is 3 seconds/drop, and stirring and reacting for 1.5 hours after the dropwise adding is finished. And then, evaporating the solvent by rotary evaporation, taking dichloromethane as an eluent, and carrying out silica gel column chromatography separation and purification on the target product.

And finally, recrystallizing in dichloromethane and methanol, wherein the volume ratio of the dichloromethane to the methanol is 1: 120 to obtain a purple solid product H₂TPP-(SH)₄The mass was 27mg, the amount of substance was 0.025mmol, and the yield was 23%.

Example 3

Tetramercaptoporphyrin H₂TPP-(SH)₄The preparation of (1):

(2)H₂TPP-(SH)₄The preparation of (1):

h is to be₂TPP-(COOH)₄(79mg, 0.10mmol) was dissolved in 4mL of dry dichloromethane, and SOCl was added in the absence of light₂(4mL, 0.069mol) was slowly added dropwise to the mixed solution, and after refluxing at 43 ℃ for 4.5 hours, excess SOCl was distilled off under normal pressure₂And dichloromethane to obtain the tetra-acyl chloride phenyl porphyrin H in black green solid state₂TPP-(COCl)₄. 83mg of H₂TPP-(COCl)₄Dissolved in 15mL of dried THF to give a first reaction solution.

1, 2-dimercaptoethane (504. mu.L, 6.0mmol) and triethylamine (40. mu.L, 0.29mmol) were dissolved in 13mL of dry THF to obtain a second reaction solution.

And dropwise adding the first reaction solution into the second reaction solution at normal temperature in a dark condition, wherein the dropwise adding speed is 3 seconds/drop, and stirring and reacting for 2.5 hours after the dropwise adding is finished. And then, evaporating the solvent by rotary evaporation, taking dichloromethane as an eluent, and carrying out silica gel column chromatography separation and purification on the target product.

Finally, recrystallizing in dichloromethane/methanol, wherein the volume ratio of dichloromethane to methanol is 1: 110 to obtain a purple solid product H₂TPP-(SH)₄The mass was 26mg, the amount of substance was 0.024mmol, and the yield was 24%.

Example 4

UiO-66-H₂TPP-(SH)₄The preparation of (1):

(1) preparation of metal organic framework material UiO-66

Weighing ZrCl₄(18.6mg, 0.080mmol), adding 3.2mL of DMF, adding 240 mu L of acetic acid, adding terephthalic acid (13.3mg, 0.080mmol), performing ultrasonic treatment to completely dissolve the acetic acid, placing the mixture into a 20mL reaction kettle, keeping the temperature at 120 ℃ for 24h, cooling to room temperature for 2h, performing high-speed centrifugal separation to obtain a solid product A, immersing the solid product A in the DMF, stirring and soaking at 80 ℃ for 6h, and performing high-speed centrifugal separation to obtain a solid product B. And immersing the solid product C in absolute ethyl alcohol, stirring and immersing for 10 hours at the temperature of 70 ℃, and performing high-speed centrifugal separation to obtain a solid product D. And continuously immersing the solid product D into absolute ethyl alcohol, stirring and immersing for 8 hours at the temperature of 65 ℃, and performing high-speed centrifugal separation to obtain a solid product E. Finally, the solid E is immersed in ether and washed, and the white powder F is obtained by airing under natural conditions, namely UiO-66, the mass of the white powder F is 14mg, the amount of the substance is 0.0084mmol, and the yield is 63%.

(2)UiO-66-H₂TPP-(SH)₄Preparation of

H from example 1 was weighed₂TPP-(SH)₄1.0 mmol. multidot.L^-1H₂TPP-(SH)₄20mg of UiO-66 was dispersed in 7.4mL of the above dichloromethane solution with sonication, and stirred at 25 ℃ for 24 hours. Centrifuging at 12500 rpm to obtain solid product G, and centrifugingWashing the solid product G with dichloromethane, immersing the solid product G in dichloromethane, stirring for 6H while immersing at normal temperature, and performing high-speed centrifugal separation to obtain a brownish red solid product UiO-66-H₂TPP-(SH)₄。

The products UiO-66 and UiO-66-H obtained in this example were characterized and analyzed₂TPP-(SH)₄The results are shown in fig. 5 to 12.

FIG. 5 is UiO-66 and UiO-66-H₂TPP-(SH)₄The left image shows that the average particle size of UiO-66 is 200nm, the surface is smooth and the shape is regular and ordered octahedron; the right picture is UiO-66-H₂TCPP-(SH)₄The reason why the octahedral morphology is basically maintained is that the tetrakismercaptoporphyrin molecule can be connected to the surface of UiO-66 by the coordination between dimercaptoethane group and zirconium cluster, the maximum pore diameter of UiO-66 is ca.0.7nm, which is much smaller than ca.1.8nm × 1.8.8 nm of porphyrin molecule, and it can be seen that the method of post-modification synthesis is only to modify porphyrin molecule to the surface of UiO-66 framework material rather than to the internal pore.

By UiO-66-H₂TPP-(SH)₄With a Transmission Electron Microscope (TEM) (fig. 6), it can be seen that the nanoparticles modified with the tetramercaptoporphyrin molecules are not connected together and are monodisperse octahedral particles.

FIG. 7 shows UiO-66 and UiO-66-H₂TPP-(SH)₄The powder diffraction Pattern (PXRD) of (1), from which can be seen, the nano material UiO-66-H modified by the tetrakismercapto porphyrin₂TPP-(SH)₄The internal structure of the UiO-66 metal organic framework material before modification is still maintained.

To further test the Tetramercaptoporphyrin modified UiO-66-H₂TPP-(SH)₄Stability in aqueous solution of UiO-66-H₂TPP-(SH)₄The samples were soaked in Phosphate Buffered Saline (PBS) at pH 7.4, stirred for 24h, centrifuged at high speed, the solids washed with copious amounts of absolute ethanol and ether, and tested for powder diffraction Pattern (PXRD) after soaking, as shown in fig. 8. As can be seen from FIG. 8, the PXRD data remained consistent before and after soakingIllustrates the tetrathiol porphyrin modified UiO-66-H₂TPP-(SH)₄Good stability in PBS.

Furthermore, from the thermogravimetric analysis (TGA) of FIG. 9, UiO-66-H₂TPP-(SH)₄Has good thermal stability as UiO-66, and the thermal stability reaches more than 400 ℃.

FIG. 10 shows UiO-66 and UiO-66-H₂TPP-(SH)₄The nitrogen adsorption and desorption tests at 77K show that UiO-66 and UiO-66-H₂TPP-(SH)₄The adsorption of nitrogen belongs to classical I-type adsorption, and the adsorption and desorption of nitrogen are reversible, further proving that the internal structural integrity of UiO-66 is not damaged by coordination reaction in the post-modification process. UiO-66-H₂TPP-(SH)₄BET specific surface area of 926m²·g^-1Specific surface area with UiO-66 of 1361m²·g^-1There is a certain reduction in comparison, which indicates that H₂TPP-(SH)₄Successfully carries out coordination modification on the surface of UiO-66 through chemical bonds.

FIG. 11 shows UiO-66 and UiO-66-H₂TPP-(SH)₄The pore size distribution of (A) shows that the tetrakismercaptoporphyrin modified UiO-66-H₂TPP-(SH)₄Has a pore size of

And

pore size distribution with UiO-66

And

in agreement, further indicating that the post-modification synthesis is only surface modification and not H₂TPP-(SH)₄Encapsulated or connected inside the aperture of the UiO-66.

The free porphyrin molecule has strong Soret band characteristic absorption characteristic at about 420nm, and has been successfully modified for determining the porphyrin moleculeOn the UiO-66 nano material, using DMF as solvent to measure UiO-66 and H₂TPP-(SH)₄And UiO-66-H₂TPP-(SH)₄Ultraviolet absorption spectrum (UV-vis), as shown in FIG. 12. As can be seen from FIG. 12, the characteristic absorption peak of UiO-66 is 265nm, H₂TPP-(SH)₄The Soret band characteristic absorption peak is 419nm, and the nano material UiO-66-H modified by tetrathioporphyrin₂TPP-(SH)₄Not only the original characteristic absorption peak of the UiO-66 nano material at 265nm is kept, but also the characteristic absorption peak of the tetrasulfhydryl porphyrin at 419nm is increased. Thus, H can be further explained₂TPP-(SH)₄Has been successfully modified on the nano material of UiO-66. To determine the content of mercaptoporphyrin, UiO-66-H was further determined₂TPP-(SH)₄C, H, N, S elemental analysis of the sample, see Table 1, for UiO-66-H₂TPP-(SH)₄The molecular formula of the compound can be expressed as [8.1 · (C)₄₈H₂₈O₃₂Zr₆)·(C₅₆H₄₅N₄O₄S₈)]。

TABLE 1UiO-66-H₂TPP-(SH)₄Elemental analysis data results

	N	C	H	S
					UiO-66-H₂TPP-(SH)₄	0.34	32.08	2.2	2.685

As a result of the above analysis, it was found that the tetrasulfhydryl porphyrin H obtained in example 1 of the present invention was used₂TPP-(SH)₄Modification of UiO-66 allows the attachment of a tetrakismercaptoporphyrin molecule to the surface of UiO-66 rather than to the internal cavity. And the modified nano material UiO-66-H₂TPP-(SH)₄The internal structure of the UiO-66 metal organic framework material before modification is still maintained, and the material has better dispersibility and stability in aqueous solution and better thermal stability. Simultaneously modified nano material UiO-66-H₂TPP-(SH)₄The porphyrin has the photosensitive characteristic maintained and may be used as one new kind of anticancer photosensitive material for PDT.

Example 5

(1) Preparation of metal organic framework material UiO-66

Weighing ZrCl₄(18.6mg, 0.080mmol), adding 4.6mL of DMF, adding 160 mu L of acetic acid, adding terephthalic acid (13.3mg, 0.080mmol), performing ultrasonic treatment to completely dissolve the acetic acid, putting the mixture into a 20mL reaction kettle, keeping the temperature at 90 ℃ for 48h, cooling to room temperature for 2h, performing high-speed centrifugal separation to obtain a solid product A, immersing the solid product A in the DMF, stirring and soaking at 60 ℃ for 10h, and performing high-speed centrifugal separation to obtain a solid product B. And then immersing the solid product B in absolute ethyl alcohol, stirring and soaking for 12 hours at the temperature of 60 ℃, and performing high-speed centrifugal separation to obtain a solid product C. And immersing the solid product C in absolute ethyl alcohol, stirring and immersing for 10 hours at the temperature of 60 ℃, and performing high-speed centrifugal separation to obtain a solid product D. Continuously immersing the solid product D into absolute ethyl alcohol, stirring and soaking for 8 hours at the temperature of 60 ℃, and performing high-speed centrifugal separation to obtain a solid product E; finally, the solid E is immersed in ether and washed, and the white powder F is obtained by airing under natural conditions, namely UiO-66, the mass of the white powder F is 14mg, the amount of the substance is 0.0084mmol, and the yield is 63%.

(2)UiO-66-H₂TPP-(SH)₄Preparation of

Weighing example 1Obtained H₂TPP-(SH)₄1.0 mmol. multidot.L^-1H₂TPP-(SH)₄20mg of UiO-66 was dispersed in 8mL of the above dichloromethane solution with sonication, and the mixture was stirred at 25 ℃ for 25 min. The solid product G was obtained by high-speed centrifugation at 12000 rpm, and was washed with methylene chloride after centrifugation. Then immersing the solid product G in dichloromethane, stirring for 5 hours at the lower side of normal temperature, and performing high-speed centrifugal separation to obtain a brownish red solid product UiO-66-H₂TPP-(SH)₄。

Example 6

(1) Preparation of metal organic framework material UiO-66

Weighing ZrCl₄(28mg, 0.12mmol), adding 5.4mL of DMF, adding 240 mu L of acetic acid, adding terephthalic acid (20.0mg, 0.12mmol), performing ultrasonic treatment to completely dissolve the terephthalic acid, putting the mixture into a 20mL reaction kettle, keeping the temperature at 120 ℃ for 12h, cooling to room temperature for 2h, performing high-speed centrifugal separation to obtain a solid A, immersing the solid product A in the DMF, stirring and immersing at 85 ℃ for 6h, performing high-speed centrifugal separation to obtain a solid product B, immersing the solid product B in absolute ethyl alcohol, stirring and immersing at 75 ℃ for 10h, and performing high-speed centrifugal separation to obtain a solid product C. And immersing the solid product C in absolute ethyl alcohol, stirring and immersing for 8 hours at the temperature of 75 ℃, and performing high-speed centrifugal separation to obtain a solid product D. Continuously immersing the solid product D into absolute ethyl alcohol, stirring and immersing for 8 hours at the temperature of 75 ℃, and performing high-speed centrifugal separation to obtain a solid product E; finally, the solid E is immersed in ether and washed, and the white powder F, namely the UiO-66, is obtained by airing under natural conditions, the mass of the white powder F is 21mg, the amount of the substance is 0.013mmol, and the yield is 65%.

(2)UiO-66-H₂TPP-(SH)₄Preparation of

H from example 1 was weighed₂TCLPP-(SH)₁1.0 mmol. multidot.L^-1H₂TCLPP-(SH)₁30mg of UiO-66 was dispersed in 6mL of the solution by sonication, and the mixture was stirred at 25 ℃ for 20 min. The solid product G is obtained by high-speed centrifugal separation, the centrifugal speed is 13000 r/min, and after centrifugation, the solid product G is washed by dichloromethane. Then will be fixedImmersing the product G in dichloromethane, stirring for 7H while immersing at normal temperature, and performing high-speed centrifugal separation to obtain a brownish red solid product UiO-66-H₂TCLPP-(SH)₁。

Example 7

Testing for singlet oxygen production

In the test of singlet oxygen generation, DMF is taken as a solvent, 1, 3-diphenyl isobenzofuran (DPBF) is taken as a capture agent of singlet oxygen, and the concentration is 60 mu mol.L^-1DPBF and 5. mu. mol. L of^-1Sample H of (1)₂TPP-(SH)₄The mixed solution is placed in a quartz cuvette and the adopted wavelength is 660nm, and the light intensity density is 20mW/cm²The laser of (1) was irradiated, and the change in absorbance of the characteristic absorption peak of DPBF at 415nm was measured every 20s time interval. As the absorption peak at 415nm disappears after the DPBF captures the singlet oxygen, the content of the singlet oxygen in the solution can be analyzed and detected according to the change of the DPBF at 415nm peak intensity. In addition, 60. mu. mol. L was taken^-1The DPBF of (1) was placed in a quartz cuvette and irradiated with a laser under the same conditions, and the change in absorbance of the characteristic absorption peak was observed as a reference. The influence of the porphyrin's UV absorption was subtracted with the same concentration of sample as background prior to detection of the UV absorption spectrum of the porphyrin-containing sample.

FIG. 13 shows that the DPBF shows little change in the peak intensity at 415nm under the same test conditions. FIG. 14 shows the addition of H₂TPP-(SH)₄The peak intensity of the post-DPBF at 415nm was very significantly reduced, indicating that the tetrakismercaptoporphyrin compound is an excellent photosensitizer.

Similarly, the concentration was taken to be 60. mu. mol. L^-1DPBF and 5. mu. mol. L of^-1(according to UiO-66-H)₂TPP-(SH)₄Molecular formula of (1) calculated as the mass concentration of porphyrin molecules) was prepared₂TPP-(SH)₄The sample was placed in a quartz cuvette and irradiated with a laser under the same conditions, and the change in absorbance of the characteristic absorption peak was observed as a reference. Before the detection of the UV absorption spectrum of the porphyrin-containing sample, the influence of the UV absorption of the porphyrin was subtracted with the same concentration of the sample as the background. As shown in FIG. 15, it can be seen that UiO-66-H was added₂TPP-(SH)₄The peak intensity of the post-DPBF at 415nm decreased very rapidly, indicating that UiO-66-H₂TPP-(SH)₄Has the same photosensitive property as the tetrathioporphyrin. And the material overcomes the problems of difficult water solubility, easy agglomeration, poor stability and the like of porphyrin molecules, and is an excellent anticancer photosensitive nano material.

The above description is only a preferred embodiment of the present application and is not intended to limit the present application, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. Preparation of UiO-66-H by modifying UiO-66 with tetrathioporphyrin₂TPP-(SH)₄The method of (2), characterized by: the method comprises the following steps:

(c) centrifuging the mixture obtained in the step (b) to obtain a solid product, washing the solid product with an organic solvent, activating the solid product, and drying to obtain the tetrakismercaptoporphyrin modified UiO-66, namely UiO-66-H₂TPP-(SH)₄；

In the step (c), the activation refers to immersing the solid product into dichloromethane, and stirring for 5-7 hours while soaking at normal temperature; the structural formula of the tetrathiol porphyrin compound is as follows:

the molecular formula of the tetra-sulfhydryl porphyrin compound is as follows: h₂TPP-(COSC₂H₄SH)₄5,10,15, 20-tetrakis (4- (S-2-mercaptoethyl) thiocarbonylphenyl) porphyrin, abbreviated as tetrakismercaptoporphyrin, H₂TPP-(SH)₄In which H is₂TPP is tetraphenylporphyrin.

2. The method of claim 1, wherein: in the step (a), the organic solvent is dichloromethane, trichloromethane and toluene;

in the step (a), the concentration of the substance in the tetrakismercaptoporphyrin solution is 1 mmol.L^-1；

In the step (b), the mass-to-volume ratio of the UiO-66 to the tetrakismercaptoporphyrin solution is 2.5-5.0: 1; the unit is mg/mL;

in the step (b), the ultrasonic dispersion time is 20-25 min;

in the step (c), the centrifugal speed is 12000-13000 r/m.

3. A preparation method of tetrasulfhydryl porphyrin is characterized in that: the method specifically comprises the following steps:

(1) h is to be₂TPP-(COOH)₄Dissolving in an organic solvent, adding thionyl chloride for refluxing, evaporating the solvent after refluxing is finished, and adding dried tetrahydrofuran to obtain a first reaction solution;

(3) dropwise adding the second reaction solution into the first reaction solution, and stirring and reacting for 1.5-3 h at room temperature; h₂TPP-(COOH)₄The mass ratio of 1, 2-dimercaptoethane to 1: 60-80 parts;

(4) subjecting the solution obtained by the reaction in the step (3) to post-treatment to obtain H in claim 1 or 2₂TPP-(SH)₄。

4. The method of claim 3, wherein: in the step (1), the hydrogen atom₂TPP-(COOH)₄The mass-volume ratio of the organic solvent to the thionyl chloride is 15-20: 1: 1; the unit is mg/mL;

in the step (1), the organic solvent is dichloromethane;

in the step (1), the reflux conditions are as follows: the temperature is 40-45 ℃, and the reflux time is 4-5 h;

in the step (1), the hydrogen atom₂TPP-(COOH)₄The mass-to-volume ratio of the tetrahydrofuran to the tetrahydrofuran is 5-7: 1; the unit is mg/mL;

in the step (2), the volume ratio of the 1, 2-dimercaptoethane to triethylamine is 10-20: 1;

in the step (2), the volume ratio of the 1, 2-dimercaptoethane to tetrahydrofuran is 0.02-0.06: 1;

in the step (3), the dripping speed is 2-3 seconds per drop, and the dripping process is carried out under the conditions of normal temperature and light shielding;

5. The method of claim 4, wherein: the column chromatography adopts silica gel column chromatography, and dichloromethane is used as eluent; the recrystallization is carried out in a solvent of dichloromethane and methanol, and the volume ratio of the dichloromethane to the methanol is 1: 100 to 120.