CN112876445A - Preparation and application of glutathione activated photosensitizer - Google Patents
Preparation and application of glutathione activated photosensitizer Download PDFInfo
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Abstract
The invention designs a photosensitizer activated by glutathione, which respectively synthesizes oxo-photosensitizer and sulfo-photosensitizerERAODI‑DNB，ASDIDNB), which not only has good PDT effect under normoxic conditions, but also exhibits excellent PDT capability under hypoxic conditions. The oxo-photosensitizer has excellent PDT effect and fluorescence emission, and excellent endoplasmic reticulum targeting function. Compared with an oxo photosensitizer, the photosensitivity of the thio photosensitizer is greatly improved by adding the S atom. After esterification with 2, 4-dinitrobenzenesulfonyl chloride, the absorption in the visible region disappeared and the photosensitivity was suppressed. After activation by glutathione, absorption in the visible region is restored, and photosensitivity is also restored. The GSH activated photosensitizer provided by the invention solves the problem of the existing photosensitizer to a certain extentThe hypoxia dependence of the preparation, low tumor specificity and the like.
Description
Technical Field
The invention belongs to the field of photodynamic therapy, and particularly relates to preparation and application of a novel photosensitizer activated by GSH.
Background
Photodynamic therapy (PDT) is an effective method for treating cancer, which is a non-invasive therapy for anti-tumor effects by generating Reactive Oxygen Species (ROS) having cytotoxicity, rapidly interacting with biological substances, and causing cell death, vascular damage, etc. However, most of the Photosensitizers (PSs) available are strongly dependent on oxygen, which severely limits the development of PDT. In addition to the anti-tumor hypoxic properties, the low tumor selectivity of photosensitizers is also an urgent problem to be solved. To improve the selectivity of the response, the development of activatable photosensitizers with stimulatory reactivity is one of the most attractive strategies. An activatable photosensitizer which can maintain the killing effect of PSs on tumor cells and avoid or reduce phototoxicity on normal tissue cells is produced. PDT can be subjected to space-time control by adjusting illumination time and illumination position, PSs are activated after reacting with specific biomolecules over-expressed by tumor cells, and ROS can be generated only by adopting light irradiation, so that the tumor cells are killed.
Glutathione is the most abundant reducing agent in biological systems and plays a crucial role in maintaining the redox balance of cells. Compared with normal tissue or cell, the cell GSH level in the tumor tissue is very high [1-10mM ], and the method provides guarantee for screening or activating tumor drugs and the like. The majority of the reported GSH-activated small organic molecule photosensitizers are type II photosensitizers, which have photodynamic action only under normoxic conditions and are effective photodynamic action under hypoxic conditions. Thus, at present, for PDT to be effective under hypoxic conditions, PS molecules capable of generating different ROS are urgently needed.
Disclosure of Invention
The invention designs a photosensitizer activated by GSH, which respectively synthesizes O-substituted photosensitizerERAODIDNB) and a photosensitizer of the S generation (A)SDIDNB), which not only has good PDT effect under normoxic conditions, but also exhibits excellent PDT capability under hypoxic conditions. The O-generation photosensitizer has excellent PDT effect and fluorescence emission and excellent endoplasmic reticulum targeting function. Compared withERAODIDNB, addition of S atom such that ASDIThe photosensitivity of DNB is greatly improved. After the photosensitizer is esterified with 2, 4-dinitrobenzenesulfonyl chloride, there is no absorption in the visible region and the photosensitivity is also inhibited. When activated by GSH, its absorption in the visible region and its fluorescence emission in the near infrared region are restored, and the photosensitizing properties are also restored. The GSH activated photosensitizer provided by the invention solves the technical problems of oxygen dependence, low tumor specificity and the like of the existing photosensitizer to a certain extent.
The present invention provides a photosensitizer activated by GSH, havingA compound of the structure shown in figure 1 of the formula specificationERAODI-DNB and ASDI-DNB。
The invention provides a preparation method of a photosensitizer activated by GSH, which is a compound shown in figure 7ERAODIStirring with anhydrous potassium carbonate at room temperature for 20min, transferring to ice water bath, cooling and stirring for 5min, adding 2, 4-dinitrobenzenesulfonyl chloride solution in dichloromethane dropwise under the protection of argon gas, monitoring by TLC for about 30min, and reacting to obtain the compound with structure shown in FIG. 1ERAODI-DNB。
The invention provides a preparation method of a photosensitizer activated by GSH, which is prepared from a compound A shown in figure 7SDIStirring with triethylamine under ice water bath condition for 10min, then adding dichloromethane solution of 2, 4-dinitrobenzene sulfonyl chloride drop by drop under argon protection condition, stirring for 30min, and then stirring at room temperature for 3 h. The reaction is completed to obtain a compound A with the structure shown in figure 1SDI-DNB。
The invention has the characteristics that:
(1) the photosensitizer is inhibited in photodynamic ability in an unactivated state, and photosensitivity is recovered after GSH reaction activation.
(2) Fluorescence emission also resumes at full rate after activation of the photosensitizer described in the present invention.
(3) The photosensitizer of the inventionERAODIAfter DNB activation, the absorption maximum is 554nm, the emission maximum is 660nm, and the Stokes shift is up to 106 nm. A. theSDIAfter DNB activation, the absorption maximum is 603nm, the emission maximum is 730nm, and the Stokes shift is up to 127 nm. The large Stokes displacement effectively reduces background interference, and the energy loss is less, thereby improving the photodynamic ability.
(4) The photosensitizer of the inventionERAODI-DNB and ASDIAfter DNB activation, tumor cells can be effectively killed under the condition of normoxic hypoxia, and hypoxic microenvironment is obtained by incubating cells by using a hypoxic chamber.
(5) The photosensitizer of the inventionERAODIExcellent endoplasmic reticulum targeting function after DNB cell uptake.
Drawings
FIG. 1 photosensitizersERAODI-DNB，ASDIChemical structure of DNB.
FIG. 2 photosensitizersERAODI-DNB，ASDI-change in absorption spectrum before and after DNB stimulus response.
FIG. 3 photosensitizersERAODI-DNB，ASDIFluorescence emission spectra before and after DNB stimulus response.
FIG. 4 photosensitizersERAODI-DNB，ASDIComparison of photosensitivity before and after DNB stimulus response, using QDPBF as indicator of active oxygen.
FIG. 5 photosensitizersERAODI-DNB，ASDIKilling effect of DNB on tumor cells under normoxic hypoxic conditions (MTT).
FIG. 6 endoplasmic reticulum-targeted photosensitizerERAODICo-localization effect of DNB in tumor cells with commercial endoplasmic reticulum probes.
FIG. 7 photosensitizersERAODIAnd ASDIThe chemical structure of (1).
FIG. 8 photosensitizersERAODI-DNB and ASDI-synthetic route to DNB.
Detailed Description
The invention will be further explained and illustrated with reference to the drawings and the specific embodiments, which are not to be considered as limiting the invention, but other embodiments obtained without inventive labour results are within the scope of protection of the invention.
The first embodiment is as follows: synthesis ofERAODI-DNB
ERAODI(0.325g,0.50mmol) was dissolved in dichloromethane (10mL) and anhydrous potassium carbonate (0.103g, 0) was added.75mmol) of the aromatic diamine, stirring the mixture for 20min at room temperature, transferring the mixture to an ice water bath, cooling and stirring the mixture for 5min, then dropwise adding 2mL of a dichloromethane solution of 2, 4-dinitrobenzenesulfonyl chloride (0.20g and 0.75mmol) under the protection of argon, stirring the mixture for 20-30min under the condition of the ice water bath, and detecting whether the reaction is finished or not by TLC. After the reaction is finished, removing the solvent by a vacuum rotary evaporator, and purifying by column chromatography to obtain a yellow solid compoundERAODI-DNB(0.28g，63.6％)。1H NMR(400MHz, DMSO-d6,)δ(ppm)9.15(d,J＝2.3Hz,1H),8.72(dd,J＝8.7,2.3Hz,1H),8.64(s,1H),8.51(d,J ＝8.7Hz,1H),8.29(s,2H),7.68–7.52(m,2H),7.25(s,1H),6.93(s,1H),4.02(s,3H),1.39(s,9H). 13C NMR(100MHz,CDCl3,)δ(ppm)164.35,157.01,153.23,152.40,152.08,148.30,139.78, 137.99,137.75,135.27,134.05,133.47,128.23,123.60,123.03,121.57,117.99,116.55, 110.17,108.06,101.04,93.54,58.99,56.83,55.38,36.04,29.75.
Example two: synthesis ASDI-DNB
ASDI(250mg,0.39mmol) was dissolved in dichloromethane (5mL) and Et was added3N0.78 mL, stirred for 10min under ice-water bath, then 3mL of a dichloromethane solution of 2, 4-dinitrobenzenesulfonyl chloride (125mg,0.48mmol) was added dropwise, the reaction was stirred for 30min under ice-water bath, and then stirred for 3h at room temperature. Completion of the reaction was checked by TLC. After the reaction is finished, removing the solvent by a vacuum rotary evaporator, and purifying by column chromatography to obtain a red solid compound ASDI-DNB (231mg,68.3％)。1H NMR(400MHz,CDCl3)δ(ppm)8.93(s,1H),8.78(s,1H),8.62(d,J＝8.8 Hz,1H),8.46(d,J＝9.1Hz,1H),8.02(s,2H),7.72(d,J＝8.5Hz,1H),7.68–7.53(m,2H),7.07(q, J＝16.0Hz,2H),1.41(s,9H).13C NMR(100MHz,CDCl3)δ(ppm)156.00,152.67,152.25,145.06, 141.94,139.32,137.36,133.78,133.65,131.30,131.17,131.03,130.45,129.01,127.19,126.88, 125.26,125.04,123.26,120.80,116.84,115.66,91.10,77.23,35.73,31.07
To further exploreERAODIResponse of DNB to GSH in solutionIn case we useERAODIDNB (5. mu.M) and ASDIDNB (5. mu.M), GSH (2.5mM), 0.3% F127, 5% DMF, 95% PBS (pH 7.4), reacted at 37 ℃ for 4h, and the UV-vis absorption spectrum was measured as shown in FIG. 2, after the addition of GSH, the molecule was analyzed fromERAODIConversion of-DNB intoERAODI，ASDIConversion of-DNB to ASDICan be absorbed and released in visible region.
To further explore AODI-DNB and ASDIResponse of DNB to GSH in solution, we use ASDIDNB (5. mu.M) and ASDIDNB (5. mu.M), GSH (2.5mM), test solution system 0.3% F127, 5% DMF, 95% PBS (pH 7.4, 10mM), reacted at 37 ℃ for 4h, and the fluorescence emission spectrum was measured. As shown in FIG. 3, after the addition of GSH, the molecules are formed byERAODIConversion of-DNB intoERAODI，ASDIConversion of-DNB to ASDIAnd the fluorescence emission in the near infrared region is released.
To further explore the photosensitizers activated by GSHERAODI-DNB and ASDIPhotodynamic status of DNB before and after GSH stimulus response, it was quantitatively tested using water-soluble 1, 3-diphenylisobenzofuran (Q-DPBF) as the active oxygen probe. The test light source is a xenon lamp, the wavelength range is 490-700nm,ERAODIDNB (2.0. mu.M), light source power 2 mW/cm2；ASDIDNB (2.0. mu.M), light source power 5mW/cm2The photodynamic ability of the test solution system of 0.3% F127, 5% DMF and 95% PBS (pH 7.4 and 10mM) before and after stimulus response is shown in figure 4, the absorption of Q-DPBF before stimulus response and the control group (without photosensitizer) are not obviously changed, and the change of Q-DPBF is rapidly increased after stimulus response, which indicates that the photodynamic of the photosensitizer before stimulus is completely inhibited and the photodynamic is recovered after stimulus.
To further explore the photosensitizer in the normal oxygenAnd the killing effect on tumor cells under the hypoxic condition, and the MTT method is utilized to researchERAODI-DNB and ASDI-DNB phototoxicity to Hela cells under normoxic and hypoxic conditions, hypoxic microenvironment was obtained by incubating the cells with hypoxic chambers, the results are shown in FIG. 5,ERAODI-DNB and ASDI-phototoxic effect of DNB on GSH overexpressed in tumor cellsERAODIAnd ASDIUnder the condition of normal oxygen and hypoxic, the tumor cells can be killed by illumination, and the cells without illumination basically keep good survival rate.
To further explore the photosensitizersERAODIEndoplasmic reticulum targeting ability of DNB, we performed co-localization analysis using a commercial endoplasmic reticulum green fluorescent probe with our photosensitizer, and found that the fluorescence coincidence of the two was good, with results as shown in FIG. 6, Pearson's correlation coefficient as high as 98%, indicating that our photosensitizerERAODIThe DNB photosensitizer has excellent endoplasmic reticulum targeting function.
The above description is only a preferred embodiment of the present invention and equivalents to substitutions or alterations made in accordance with the present invention (e.g., photosensitizers prepared by etherification of different GSH activating groups, respectively, with a precursor substrate for the photosensitizer) are within the scope of the present invention.
Claims (10)
1. A glutathione activated photosensitizer is a compound with a structure shown in figure 1 in the specification.
2. A method for preparing glutathione activated photosensitizer is shown in figure 7 in the specification.
3. A compound according to claim 1ERAODIAnd 2, 4-dinitrobenzenesulfonyl chloride in a molar ratio of 1: 1.5 reaction solvent is dry dichloromethane, CompoundERAODIStirring with potassium carbonate at room temperature for 20min, transferringCooling and stirring in an ice-water bath for 5min, then dropwise adding a dichloromethane solution of 2, 4-dinitrobenzenesulfonyl chloride under the protection of argon, stirring the reaction for 30min under the condition of the ice-water bath, and detecting whether the reaction is finished by TLC.
4. According to claim 1, compound ASDIAnd 2, 4-dinitrobenzenesulfonyl chloride in a molar ratio of 1: 1.2 reaction solvent is dry dichloromethane, Compound ASDIAnd triethylamine are stirred for 10min under the condition of ice-water bath, then dichloromethane solution of 2, 4-dinitrobenzenesulfonyl chloride is added drop by drop, the reaction is stirred for 30min under the condition of ice-water bath, then the reaction is stirred for 3h at room temperature, and whether the reaction is finished or not is detected by TLC.
5. According to the method of claim 1, wherein,ERAODI-DNB and ASDIPhotosensitizers of the structure shown by DNB have no photosensitizing effect when exposed to GSH
After activation, a phototoxic photosensitizer is formedERAODIAnd ASDIThe photosensitivity is restored.
6. A photosensitizer according to claim 1ERAODI-DNB and ASDIFluorescence emission also resumes at full speed after DNB is activated by GSH.
7. According to the method of claim 1, wherein,ERAODI-the maximum absorption of the DNB photosensitizer after activation by GSH is 554nm, the maximum emission is 660nm, the stokes shift is 106 nm; a. theSDIAfter the DNB photosensitizer is activated by GSH, the maximum absorption is 603nm, the maximum emission is 730nm, the Stokes shift is up to 127nm, the interference of biological background is effectively reduced, and the photodynamic capacity is improved.
8. According to the method of claim 1, wherein,ERAODI-DNB and ASDIAfter DNB activation, in normoxia and hypoxiaUnder the condition, tumor cells can be effectively killed, and a hypoxic microenvironment is obtained by incubating the cells by using a hypoxic chamber.
9. According to the method of claim 1, wherein,ERAODIafter DNB activation, the DNA has excellent endoplasmic reticulum targeting property, and the Pearson correlation coefficient is as high as 98%.
10. The method of claim 1, wherein the substitution or alteration is equivalent to that of the present strategy, and is within the scope of the present invention.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107235945A (en) * | 2017-06-08 | 2017-10-10 | 浙江工业大学 | A kind of glutathione that responds kills photaesthesia targeting anti-tumor prodrug of tumour cell and preparation method and application |
| CN108727372A (en) * | 2018-06-20 | 2018-11-02 | 华中师范大学 | A kind of fluorescence probe and preparation method thereof of quick identification glutathione |
| CN110698449A (en) * | 2019-06-12 | 2020-01-17 | 南开大学 | Preparation method and application of novel photosensitizer |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107235945A (en) * | 2017-06-08 | 2017-10-10 | 浙江工业大学 | A kind of glutathione that responds kills photaesthesia targeting anti-tumor prodrug of tumour cell and preparation method and application |
| CN108727372A (en) * | 2018-06-20 | 2018-11-02 | 华中师范大学 | A kind of fluorescence probe and preparation method thereof of quick identification glutathione |
| CN110698449A (en) * | 2019-06-12 | 2020-01-17 | 南开大学 | Preparation method and application of novel photosensitizer |
Non-Patent Citations (1)
| Title |
|---|
| MENG LI等: ""A near-infrared colorimetric fluorescent chemodosimeter for the detection of glutathione in living cells"", 《CHEM.COMMUN.》 * |
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