CN115109052B - AIE compound with mitochondrial targeting and synthesis method and application thereof - Google Patents
AIE compound with mitochondrial targeting and synthesis method and application thereof Download PDFInfo
- Publication number
- CN115109052B CN115109052B CN202210819688.0A CN202210819688A CN115109052B CN 115109052 B CN115109052 B CN 115109052B CN 202210819688 A CN202210819688 A CN 202210819688A CN 115109052 B CN115109052 B CN 115109052B
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- aie
- mitochondrial
- triphenylamine
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Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
- C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
- C07D417/06—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
- A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1003—Carbocyclic compounds
- C09K2211/1011—Condensed systems
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1003—Carbocyclic compounds
- C09K2211/1014—Carbocyclic compounds bridged by heteroatoms, e.g. N, P, Si or B
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1029—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1044—Heterocyclic compounds characterised by ligands containing two nitrogen atoms as heteroatoms
- C09K2211/1051—Heterocyclic compounds characterised by ligands containing two nitrogen atoms as heteroatoms with sulfur
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
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Abstract
The invention discloses an AIE compound with mitochondrial targeting, which enhances the charge intensity of the compound by combining molecules with aggregation-induced emission structures and long-chain molecules with positive charges, and the obtained compound can target mitochondria of tumor cells to realize the therapeutic effects of dark toxicity and phototoxicity on the tumor cells; the compound has the functions of inducing active oxygen production in cancer cells, reducing mitochondrial membrane potential, changing mitochondrial morphology, damaging mitochondrial function and inducing autophagy of tumor cells (mitochondria); in addition, due to the inherent optical property of AIE molecules, cancer cells and normal cells can be distinguished through fluorescence imaging, so that diagnosis and treatment integration can be realized, and the method has wide application prospects. The invention also discloses a synthesis method of the AIE compound with mitochondrial targeting and application of the AIE compound in preparation of antitumor drugs.
Description
Technical Field
The invention relates to an AIE compound with mitochondrial targeting, and also relates to a synthesis method of the compound and application of the compound in preparation of antitumor drugs.
Technical Field
Mitochondria are important organelles in human cells, the outer membrane has a negative potential, and the inner membrane is rich in proteins involved in protein input mechanisms, ATP synthase and respiratory chain complexes. As a cellular motive, mitochondria synthesize the energy-rich molecule Adenosine Triphosphate (ATP) to support a variety of cellular processes, including metabolism, cell division, and biomolecular synthesis. In addition to functioning as an energy factory, mitochondria are responsible for many other important functions including initiation of apoptotic pathways, calcium metabolism, electron transfer, lipid metabolism, reactive oxygen species generation, urea cycle, citric acid cycle, and gluconeogenesis and steroid hormone synthesis.
The concept of aggregation-induced emission (AIE) was proposed by Tang Benzhong institutions et al in 2001. In this phenomenon, the molecules are strongly fluorescent when aggregated or in the solid state, but weakly when dispersed in solution. In view of their advantages, such as high light stability, large stokes shift, and ease of preparation. A wide variety of AIE luminophores (AIEgens) have been extensively explored, particularly in the fields involving bioimaging, sensing and therapy. Side chain engineering refers to the modulation of the properties of a molecule by altering the side chain structure of the molecule. In the aspect of constructing an AIE active multifunctional photochemical system, side chain engineering can be used as an effective method.
Disclosure of Invention
The invention aims to: based on the characteristic that mitochondria of tumor cells are different from normal cells in structure and function, one of the purposes of the invention is to provide an AIE compound capable of targeting mitochondria of tumor cells, the AIE compound can still keep fluorescence emission performance related to radiation transition by controlling the length of a side chain, and meanwhile, the D-A intensity of the compound can be improved by introducing the side chain, so that the generation of active oxygen in cancer cells is induced; the invention also aims to provide a synthesis method of the compound and application of the compound in preparation of antitumor drugs.
The technical scheme is as follows: the chemical structural formula of the AIE compound with mitochondrial targeting is shown as formula (I):
the synthesis method of the AIE compound with the mitochondrial targeting specifically comprises the following steps: firstly synthesizing a triphenylamine parent structure compound with an aldehyde group, and then reacting the triphenylamine parent structure compound with the aldehyde group with pyridine with a long chain to obtain an AIE compound with mitochondrial targeting;
wherein, the chemical structural formula of the triphenylamine parent structural compound with aldehyde group is as follows:
wherein in pyridine with long chain, the linear alkyl of C4-12 is grafted on one side of pyridine N atom.
Further preferably, the pyridine N atom is grafted with a C8 straight chain alkyl group, and the chemical structural formula of the pyridine with a long chain is as follows:
wherein, specifically, it is: heating and refluxing a triphenylamine parent structure compound with an aldehyde group and pyridine with a long chain in absolute ethyl alcohol, extracting, separating and extracting a crude product after the reaction is finished, and purifying by a column chromatography to obtain the AIE compound with mitochondrial targeting.
Wherein, the triphenylamine parent structure molecule with aldehyde group is prepared by the following method: heating and refluxing a compound (4, 4'- [ [4- (4, 5-tetramethyl-1, 3, 2-dioxabolan 2-ethyl) phenyl iminodi, 1' -dimethyl ester) with 7-bromo-4-aldehyde benzo [ C ] [1,2,5] thiadiazole in a mixed solution of tetrahydrofuran and water in an inert atmosphere, extracting and separating to extract a crude product after the reaction is finished, and purifying by a column chromatography to obtain the triphenylamine parent structure compound with an aldehyde group;
wherein, the chemical structural formula of the compound with triphenylamine parent structure is as follows:
7-bromo-4-aldehyde benzo [ C][1,2,5]The thiadiazole has a chemical structural formula:
wherein, the molar ratio of the triphenylamine parent structure compound with aldehyde group to the pyridine with long chain is 1:1.2; the reflux reaction time is 4-4.5 h; the reaction temperature is 80-85 ℃.
Wherein, the reaction mole ratio of the compound with triphenylamine parent structure and 7-bromo-4-aldehyde benzo [ C ] [1,2,5] thiadiazole is 1:1.2; the reflux reaction time is 18-19 h; the reaction temperature is 70-75 ℃.
The AIE compound with mitochondrial targeting is applied to the preparation of antitumor drugs and antitumor drug components.
Wherein the tumor is cervical cancer.
The chemical reaction formula of the compound is as follows:
the beneficial effects are that: the invention firstly prepares a molecule with an aggregation-induced emission structure (triphenylamine rotor structure), one end of the molecular structure is an electron-withdrawing end, and the other end is an electron-donating end, and simultaneously combines the molecule with the aggregation-induced emission structure with a long-chain molecule with positive charges, so that the charge intensity of the compound is further enhanced, the obtained compound can target mitochondria of tumor cells, and the therapeutic effects of dark toxicity and phototoxicity on the tumor cells are realized; the compound has the functions of inducing active oxygen production in cancer cells, reducing mitochondrial membrane potential, changing mitochondrial morphology, damaging mitochondrial function and inducing autophagy of tumor cells (mitochondria); in addition, due to the inherent optical property of AIE molecules, cancer cells and normal cells can be distinguished through fluorescence imaging, so that diagnosis and treatment integration can be realized, and the method has wide application prospects.
Drawings
FIG. 1 is a graph showing cellular uptake of compound TPA-N-8 of example 1;
FIG. 2 is a graph of subcellular organelle localization of compound TPA-N-8 of example 1;
FIG. 3 is a schematic representation of the compound TPA-N-8 of example 1 inducing HeLa cells to produce reactive oxygen species;
FIG. 4 is a schematic representation of the compound TPA-N-8 of example 1 inducing a decrease in mitochondrial membrane potential in HeLa cells;
FIG. 5 is a schematic representation of the effect of the compound TPA-N-8 of example 1 on mitochondrial morphology and function;
FIG. 6 is a schematic representation of the TPA-N-8 compound of example 1 inducing changes in the levels of related proteins in HeLa cells.
Detailed Description
The technical scheme of the invention is further described below with reference to specific embodiments.
Example 1
The invention relates to a synthesis method of a long-chain AIE compound (TPA-N-8) with mitochondrial targeting, which comprises the following steps:
(1) Preparation of triphenylamine parent Structure Compound with aldehyde group (TPA-CHO): compound 1 (compound with triphenylamine parent structure) (1.463 g,3 mol), compound 2 (7-bromo-4-aldyl benzo [ C ] [1,2,5] thiadiazole) (0.7533 g,3.1 mol), anhydrous potassium carbonate (4.14 g,30 mol) were weighed into a three-necked flask, isolated from air, using argon as a shielding gas, and tetrakis (triphenylphosphine) palladium (0.1155 g,0.1 mol) was added as a reaction solvent in a tetrahydrofuran/water (tetrahydrofuran/water volume ratio of 3:1), and heated to reflux at 70 ℃ for 18h. Post-treatment: stopping stirring, and removing tetrahydrofuran by rotary evaporation; adding more water, extracting for multiple times, adding anhydrous magnesium sulfate, and drying. The mixture was filtered off with suction, the solvent was removed from the filtrate by rotary evaporation, an orange solid appeared, and the solid was dissolved with dichloromethane and then stirred with silica gel. Column chromatography separation is carried out by using developing agent (volume ratio: petroleum ether: ethyl acetate=15:1-petroleum ether: ethyl acetate=5:1) to obtain a product TPA-CHO (orange solid, yield 65%), wherein the TPA-CHO has a chemical structural formula:
the chemical reaction formula of the triphenylamine parent structural compound (TPA-CHO) with aldehyde group is:
by the characterization of nuclear magnetism and mass spectrum, 1 H NMR(400MHz,DMSO-d 6 )δ(ppm):8.34(d,J＝7.4Hz,1H),8.09(dd,J＝12.0,7.9Hz,3H),7.91(d,J＝8.5Hz,4H),7.30(d,J＝8.4Hz,2H),7.17(d,J＝8.6Hz,4H),3.83(s,6H). 13 C NMR(101MHz,DMSO-d 6 )δ(ppm):189.50,166.09,153.56,153.40,150.68,147.06,138.34,133.55,132.84,131.71,131.46,127.44,126.34,125.94,124.62,123.54,52.45.ESI-MS(CH 3 OH):calcd for[TPA-CHO]m/z＝523.1，found m/z＝523.2。
(2) TPA-CHO (0.1046 g,0.2 mol) is weighed into a reaction device, vacuumized and argon is used as a protective gas; adding long chain (8-N) + ) (0.027 g,0.22 mol) absolute ethanol was used as a solvent, and after stirring at 80℃for 10 minutes to dissolve sufficiently, a small amount of piperidine (50. Mu.L) was added thereto, and the mixture was refluxed at 80℃for 4 hours. Post-treatment: stopping stirring and removing the reaction solvent by rotary evaporation; the solid was dissolved with dichloromethane and then stirred with silica gel. Column chromatography separation is carried out by using developing agent (volume ratio: dichloromethane: methanol=20:1-dichloromethane: methanol=10:1) to obtain the product TPA-N-8 (reddish brown solid, yield 40%), wherein the chemical structural formula of TPA-N-8 is as follows:
the chemical reaction formula of a long chain AIE compound with mitochondrial targeting (TPA-N-8) is:
by the characterization of nuclear magnetism and mass spectrum, 1 H NMR(400MHz，DMSO-d 6 )δ(ppm):9.04(d,J＝6.6Hz,2H),8.45–8.31(m,4H),8.19–8.10(m,3H),8.05(d,J＝7.5Hz,1H),7.93(d,J＝8.8Hz,4H),7.34–7.27(m,2H),7.21–7.15(m,4H),4.54(t,J＝7.4Hz,2H),3.84(s,6H),1.97–1.89(m,2H),1.31–1.20(m,12H),0.88–0.83(m,3H). 13 C NMR(101MHz,DMSO-d 6 )δ166.13,153.12,150.76,146.53,144.86,136.97,134.23,132.89,131.47,131.43,128.36,127.88,127.25,126.14,124.70,124.53,123.40,60.29,52.46,31.61,31.09,29.47,28.93,28.85,25.93,22.51,14.41.ESI-MS(CH 3 OH):calcd for[TPA-N-8]m/z＝711.3，found m/z＝711.4。
comparative example 1: synthesis of TPA-N-4
Comparative example 1TPA-N-4 was synthesized in the same manner as in example 1, except that the long chain (4-N + ) The structural formula of the obtained compound TPA-N-4 is as follows:
by the characterization of nuclear magnetism and mass spectrum, 1 H NMR(400MHz,Chloroform-d)δ(ppm):9.13(d,J＝6.5Hz,2H),8.34–8.25(m,3H),8.17(d,J＝16.2Hz,1H),8.07(d,J＝7.5Hz,1H),7.99–7.91(m,6H),7.75(d,J＝7.4Hz,1H),7.27(d,J＝10.6Hz,2H),7.20–7.16(m,4H),4.83(t,J＝7.4Hz,2H),3.92(s,6H),2.04(dd,J＝8.7,6.5Hz,2H),1.50–1.44(m,2H),1.00(t,J＝7.4Hz,3H). 13 C NMR(101MHz,Chloroform-d)δ(ppm):166.52,153.80,150.60,146.91,144.18,137.93,135.27,133.60,132.59,131.17,130.68,127.47,126.74,126.34,125.29,124.92,124.61,123.31,61.01,52.08,33.53,29.70,19.43,13.62,1.03.ESI-MS(CH 3 OH):calcd for[TPA-N-4]m/z＝655.2，found m/z＝655.3。
comparative example 2: synthesis of TPA-N-12
Comparative example 2TPA-N-12 was synthesized in the same manner as in example 1, except that the long chain (12-N + ) The structural formula of the obtained compound TPA-N-12 is as follows:
by the characterization of nuclear magnetism and mass spectrum, 1 H NMR(400MHz,DMSO-d6)δ(ppm):9.04(d,J＝6.6Hz,2H),8.45–8.29(m,4H),8.14(dd,J＝18.6,8.1Hz,3H),8.05(d,J＝7.5Hz,1H),7.92(d,J＝8.8Hz,4H),7.34–7.26(m,2H),7.19–7.14(m,4H),4.54(t,J＝7.3Hz,2H),3.83(s,6H),1.92(s,2H),1.30–1.20(m,20H),0.85–0.80(m,3H). 13 C NMR(101MHz,DMSO-d6)δ(ppm):166.11,153.10,150.74,146.51,144.86,136.97,133.33,132.93,131.46,131.41,128.35,127.84,126.11,124.68,124.51,123.39,60.28,52.46,31.77,31.07,29.49,29.38,29.26,29.19,28.87,25.90,22.56,14.42.ESI-MS(CH 3 OH):calcd for[TPA-N-12]m/z＝767.3，found m/z＝767.4。
test compounds TPA-N-4, TPA-N-8 and TPA-N-12, lipid partition coefficient logP o/w ：
Fat solubility of the compounds by classical shake flask method (using lipid partition coefficient log P o/w Representation) and analyzed using an ultraviolet-visible spectrometer. Deionized water is used as the water phase, n-octanol is used as the organic phase, two phases are mixed and oscillated for 24 hours to enable the water phase to be saturated with each other, the compound (100 mu M) is respectively dissolved in the water phase saturated with n-octanol, the organic phase saturated with equal volume of water is added, the water phase is oscillated for 4 hours on a shaking table at 37 ℃ and centrifuged for 5 minutes at a rotating speed of 3000rpm, the absorbance values of the compound in the water phase and the organic phase are respectively detected by an ultraviolet-visible spectrometer, and the logarithm of the ratio of the concentration of the compound in the organic phase and the water phase is obtained to obtain a lipid water distribution coefficient log P o/w Values.
The compounds TPA-N-4, TPA-N-8 and TPA-N-12 have a lipid partition coefficient logP o/w As shown in table 1.
TABLE 1
As can be seen from Table 1, the compound TPA-N-8 has slightly higher lipid solubility than the compounds TPA-N-4 and TPA-N-12, and more lipophilic molecules are more accessible to cells.
Application of compounds TPA-N-4, TPA-N-8 and TPA-N-12 to antitumor activity of cervical cancer cell HeLa:
the antiproliferative effect of compounds TPA-N (n=4/8/12) and cisplatin CDDP on cervical cancer cells HeLa was analyzed using MTT colorimetric method. In living cells, mitochondrial succinate dehydrogenase can reduce MTT to a blue-violet product: formazan (soluble in DMSO) and the product has an absorption peak at 570nm, so A can be used 570nm To analyze the proliferation of cells.
The specific experimental steps are as follows:
(1) Firstly, resuscitating a tube of tumor cells, culturing with fresh culture solution (wherein HeLa cells use DMEM culture medium+10% fetal calf serum+1% penicillin and streptomycin), and carrying out passage for 2 times;
(2) When the cells reached the logarithmic phase, they were inoculated into 96-well plates (100. Mu.L of culture medium per well) at a cell density of 5000 cells/well, followed by placing in an incubator (37 ℃,5% CO) 2 ) Culturing in medium;
(3) After the cells are attached, 100 mu L of fresh culture solution containing TPA-N-N (n=4/8/12) with different concentration gradients and cisplatin CDDP is added into each hole, and then the cells are placed in an incubator for continuous incubation;
(4) After incubation for 24 hours, 20. Mu.L MTT (5 mg/mL) was added to each well, after incubation for 4 hours in a 37℃incubator, the supernatant was aspirated, 150. Mu.L of dimethyl sulfoxide (DMSO) was added to each well, and detection of A was performed using an ELISA 570 nm Calculating the cell proliferation inhibition ratio and obtaining IC 50 Value (drug concentration corresponding to inhibition equal to 50%). The MTT test results for compounds TPA-N (n=4/8/12) and cisplatin CDDP are shown in table 2:
TABLE 2
The results show that: the proliferation inhibition activity of the compound TPA-N-8 on tumor cells is higher than that of the compounds TPA-N-4/12 and CDDP, which shows that the compound TPA-N-8 with the best lipophilic effect has good inhibition effect on cervical cancer cells HeLa; in addition, the compound TPA-N-8 not only shows better toxicity, but also can improve the treatment effect under the illumination condition; the compound TPA-N-8 still has good therapeutic effect on cervical cancer cells HeLa in the hypoxic environment, and solves the condition limitation of the tumor hypoxic environment.
Use of the compound TPA-N-8 of example 1 for uptake by HeLa cells:
the compound TPA-N-8 was tested for uptake in HeLa cells. Inoculation onto coverslips in 12 well plates 5 x 10 4 HeLa cells in logarithmic growth phase were placed in incubator for wall-attached growth overnight, and cell culture solution containing compound TPA-N-8 prepared in advance was added for incubation for different times (0.5, 1, 1.5, 2, 3, 4 h). The culture was aspirated and washed three times with PBS. The shots were then observed under a confocal microscope. Compound TPA-N-8: lambda (lambda) ex ＝488nm，λ em ＝700nm。
The uptake of TPA-N-8 prepared in example 1 in HeLa cells is shown in FIG. 1. The results show that: the lipophilic compound TPA-N-8 can completely enter cells after being incubated with HeLa for 2 hours, thereby laying a foundation for improving the anti-tumor curative effect.
Use of compounds TPA-N-4, TPA-N-8, TPA-N-12 for intracellular subcellular organelle (mitochondrial) localization:
the compounds TPA-N-4, TPA-N-8, TPA-N-12 were assayed for their distribution in the mitochondria and lysosomes in HeLa cells using commercial probes MitoGreen, lysoGreen. Inoculation onto coverslips in 12 well plates 5 x 10 4 HeLa cells in the logarithmic growth phase were placed in an incubator and grown on wall overnight, and the prepared cell culture solutions containing 5. Mu.L of the compounds TPA-N-4, TPA-N-8 and TPA-N-12 were added, respectively, and incubation was continued for 2 hours. The culture was aspirated and washed three times with PBS. Subcellular organelle staining was accomplished by adding a culture solution containing commercial probes at concentrations and incubating for a period of 30min before testing, followed by observation under confocal microscopy. Compound TPA-N-8: lambda (lambda) ex ＝488nm，λ em =700 nm; excitation and emission wavelength ranges of MitoGreen commercial probes: lambda (lambda) ex ＝488nm，λ em =525 nm; excitation and emission wavelength ranges of LysoGreen commercial probes: lambda (lambda) ex ＝405nm，λ em =525 nm. Co-localization coefficient analysis was analyzed with ImageJ software.
The confocal images of TPA-N-8 prepared in example 1 after co-incubation with mitochondrial and lysosomal probes are shown in FIG. 2. The results show that: the co-localization coefficient of TPA-N-8 with mitochondrial probe was 0.805 and with lysosomal probe was 0.259, indicating that TPA-N-8 was able to efficiently localize in mitochondria within HeLa cells.
Use of TPA-N-8 prepared in example 1 to induce intracellular reactive oxygen species formation:
method 1: confocal microscopy detects ROS within cancer cells. HeLa cells were seeded in 35mm Corning laser confocal dishes and when the cell density was grown to 70%, after 2h treatment with the compound TPA-N-84. Mu.L, the cells were stained for 30min at 37℃in a serum-free medium containing 10. Mu.M DCF-DA, immediately followed by confocal microscopy at an excitation wavelength of 488nm and an emission wavelength of 530.+ -. 20nm.
Method 2: flow cytometry detects ROS within cancer cells. HeLa cells were treated with TPA-N-8 (1/2/4/6/8. Mu.L) for 2 hours, stained with 10. Mu.M DCF-DA in serum-free culture for 30min at 37℃in the absence of light, centrifuged to discard the supernatant, and washed three times with serum-free medium to remove DCF-DA that did not enter the cells; measuring green fluorescence intensity by a flow cytometer within the second half hour of collecting the cells; the excitation wavelength was 488nm and the emission wavelength was 530.+ -.20 nm. The average fluorescence intensity of green light was analyzed using FlowJo7.6 (Tree Star, OR, USA) software.
The results of the compound TPA-N-8 on the induction of intracellular reactive oxygen species are shown in FIG. 3. The results show that: compared with the control group, the compound TPA-N-8 can effectively induce the increase of the intracellular active oxygen content after being treated, and after 6 mu L of compound TPA-N-8 is added into HeLa cells, the intracellular active oxygen substance is increased by about 10 times, and meanwhile, the green fluorescence of the confocal part of the graph of FIG. 3 is obviously enhanced. While the intracellular Reactive Oxygen Species (ROS) in the control group (DMSO only) treated group did not change significantly, indicating that the compound TPA-N-8 induced Reactive Oxygen Species (ROS) to be produced by the cancer cells after entry into the cancer cells, resulting in death of the cancer cells.
Use of TPA-N-8 prepared in example 1 to induce changes in intracellular mitochondrial membrane potential:
flow cytometry detects changes in mitochondrial membrane potential within tumor cells. Adding cell culture solution containing compound TPA-N-8 (1/2/4/6/8 μl) into 6-well plate inoculated with HeLa cells with good morphology and normal growth, treating with the drug for 2 hr, collecting cells, washing with PBS, and adding the prepared JC-1 working solution for dyeing for 25-30min; washing cells with bufferAnd re-suspended, the samples were immediately tested using a BDFACSverse flow cytometer and the results were processed and analyzed using FlowJo7.6 software. Detection of fluorescent channel as lambda ex ＝488nm，λ em ＝530nm；λ ex ＝488nm，λ em ＝590nm。
The results of the compound TPA-N-8 on inducing changes in intracellular mitochondrial membrane potential are shown in FIG. 4. The results show that: an increase in JC-1 monomer and a decrease in JC-1 aggregate can be observed after treatment with compound TPA-N-8 compared with the control group, indicating that compound TPA-N-8 effectively induces a decrease in mitochondrial membrane potential.
Effects of the compound TPA-N-8 prepared in example 1 on mitochondrial morphology and function:
mitochondrial morphology of HeLa cells after treatment with compound TPA-N-8 was observed by Transmission Electron Microscopy (TEM). 100mm to which HeLa cells had been seeded 3 Adding a pre-prepared cell culture solution containing 4 mu L of a compound TPA-N-8 into a culture dish for drug treatment, after co-incubation for 24 hours, collecting cells, washing with PBS, fixing with 2.5% glutaraldehyde, gradient dehydrating with alcohol, embedding with resin, ultrathin slicing, staining with uranyl acetate and lead citrate, preparing a sample to be observed on a copper wire, and observing the mitochondrial morphology by using a Hitachi H-7650 transmission microscope.
The experimental results of the effect of compound TPA-N-8 on mitochondrial morphology are shown in FIG. 5. The results indicate that the compound TPA-N-8 (4. Mu.M) treated HeLa cells showed a rounded mitochondrial swelling and a loss of cristae structure compared to the intact mitochondrial cristae structure of the Control group without drug treatment. This demonstrates morphologically the characteristic of the compound TPA-N-8 to damage HeLa cell mitochondria.
Use of TPA-N-8 prepared in example 1 for inducing autophagy of HeLa cells:
autophagic protein content changes were detected using Western Blotting (WB). Adding a pre-prepared cell culture solution containing a compound TPA-N-8 (2/4/8 mu L) and a control group (containing DMSO) into a 100mL culture dish for growing HeLa cells in an adherence way, centrifuging to collect cells after the drug is treated for 24 hours, washing with PBS to remove serum in the residual culture solution, adding a strong lysis solution containing PMSF (PMSF) and carrying out whole cell lysis for 20 minutes, and ensuring a low-temperature environment at 4 ℃ in the whole process to ensure protein invariance. Centrifuging at 13400rpm for 20min under low temperature condition, and sucking supernatant obtained by centrifugation to obtain cell whole protein sample required by experiment; determining the protein concentration in the protein sample using a BCA protein content assay kit; and detecting the expression content of different proteins in the sample by SDS-PAGE gel electrophoresis. After the gel is prepared, adding protein samples with the same volume into each hole for gel electrophoresis experiments, and immediately stopping electrophoresis after proper separation; the target protein was transferred onto PVDF membrane using a wet method, and after the completion, the membrane was blocked in 5% nonfat milk powder for 2 hours. Diluting the primary antibody with skimmed milk powder according to the corresponding proportion according to the instruction of the antibody, and placing the sealed membrane in a primary antibody incubation liquid for incubation for a period of time at room temperature so as to enable the membrane to be specifically combined with the target protein. After completion, the cells were washed with PBST (4X 8 min/time). The washed membrane was incubated in a pre-formulated secondary antibody incubation for a period of time to bind to the primary antibody, again washed with PBST. An equal volume of ECL developer is prepared, covered on a PVDF film, and photographed by a chemiluminescent imaging system after 2min of treatment.
The experimental results of the compound TPA-N-8 on HeLa cells to induce the expression of cell autophagy-related proteins are shown in FIG. 6. The results show that: compared with a control group which is not treated by adding medicine, after the cells are treated by TPA-N-8, the expression of the mitochondrial autophagy related protein PINK1 is obviously up-regulated, and meanwhile, the content of autophagy marker protein LC3-II is increased in a concentration-dependent manner, which indicates that the death mode of HeLa cells induced by the compound TPA-N-8 is autophagy.
Claims (8)
1. An AIE compound having mitochondrial targeting, wherein the chemical structural formula of the compound is shown in formula (I):
2. the method of synthesis of a mitochondrial-targeted AIE compound of claim 1, wherein the method is: firstly synthesizing a triphenylamine parent structure compound with an aldehyde group, and then reacting the triphenylamine parent structure compound with the aldehyde group with pyridine with a long chain to obtain an AIE compound with mitochondrial targeting;
wherein, the chemical structural formula of the triphenylamine parent structural compound with aldehyde group is as follows:
wherein in the pyridine with long chain, C8 straight chain alkyl is grafted on one side of pyridine N atom, and the chemical structural formula of the pyridine with long chain is as follows:
3. the method of synthesis of AIE compounds with mitochondrial targeting according to claim 2, characterized in that it is in particular: heating and refluxing a triphenylamine parent structure compound with an aldehyde group and pyridine with a long chain in absolute ethyl alcohol, extracting, separating and extracting a crude product after the reaction is finished, and purifying by a column chromatography to obtain the AIE compound with mitochondrial targeting.
4. The method of synthesizing a mitochondrial-targeted AIE compound of claim 2, wherein the triphenylamine parent structural molecule having an aldehyde group is prepared by the following method: heating and refluxing a compound with a triphenylamine parent structure and 7-bromo-4-aldehyde benzo [ C ] [1,2,5] thiadiazole in a mixed solution of tetrahydrofuran and water in an inert atmosphere, extracting, separating and extracting a crude product after the reaction is finished, and purifying by a column chromatography to obtain the triphenylamine parent structure compound with aldehyde groups;
wherein, the chemical structural formula of the compound with triphenylamine parent structure is as follows:
5. the method of synthesis of a mitochondrial-targeted AIE compound according to claim 3 wherein: the molar ratio of the triphenylamine parent structure compound with aldehyde group to the pyridine with long chain is 1:1.2; the reflux reaction time is 4-4.5 h; the reaction temperature is 80-85 ℃.
6. The method of synthesis of AIE compounds with mitochondrial targeting according to claim 4, wherein: the reaction mole ratio of the compound with triphenylamine parent structure and 7-bromo-4-aldehyde benzo [ C ] [1,2,5] thiadiazole is 1:1.2; the reflux reaction time is 18-19 h; the reaction temperature is 70-75 ℃.
7. Use of the AIE compound with mitochondrial targeting according to claim 1 for the preparation of antitumor drugs and antitumor drug fractions.
8. The use of the AIE compound with mitochondrial targeting according to claim 7 for the preparation of antitumor drugs and antitumor drug fractions, characterized in that: the tumor is cervical cancer.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110642852A (en) * | 2019-10-25 | 2020-01-03 | 南开大学 | Organic AIE photosensitive probe with mitochondrion targeting function and preparation method and application thereof |
| CN111690005A (en) * | 2020-06-27 | 2020-09-22 | 北京化工大学 | Sensing array with mitochondrial targeting and aggregation-induced emission effects and application of sensing array in cell identification |
| CN113956292A (en) * | 2021-10-29 | 2022-01-21 | 南京师范大学 | Mitochondrial targeting compound with aggregation-induced emission property and synthesis method and application thereof |
| CN114195774A (en) * | 2021-11-04 | 2022-03-18 | 徐州医科大学 | Photosensitizer with hypochlorous acid activated fluorescence and mitochondrion targeting functions and preparation method and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110642852A (en) * | 2019-10-25 | 2020-01-03 | 南开大学 | Organic AIE photosensitive probe with mitochondrion targeting function and preparation method and application thereof |
| CN111690005A (en) * | 2020-06-27 | 2020-09-22 | 北京化工大学 | Sensing array with mitochondrial targeting and aggregation-induced emission effects and application of sensing array in cell identification |
| CN113956292A (en) * | 2021-10-29 | 2022-01-21 | 南京师范大学 | Mitochondrial targeting compound with aggregation-induced emission property and synthesis method and application thereof |
| CN114195774A (en) * | 2021-11-04 | 2022-03-18 | 徐州医科大学 | Photosensitizer with hypochlorous acid activated fluorescence and mitochondrion targeting functions and preparation method and application thereof |
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