Aggregation-induced luminescence photosensitizer that simultaneously generates type I and type II reactive oxygen species and its application

Abstract

Aggregation-induced luminescence photosensitizers that simultaneously generate type I and type II reactive oxygen species and their applications relate to the field of chemistry. Aggregation-induced luminescence photosensitizers that simultaneously generate type I and type II reactive oxygen species can simultaneously generate type I and type II reactive oxygen species, and the amount of reactive oxygen species generated is much higher than that of commercial dyes, with better photodynamic effects. The present invention effectively improves the environmental limitations of photodynamic therapy and is expected to replace commercial photosensitizers to achieve multiple applications in multiple occasions, such as antibacterial and anti-tumor in vivo, and antibacterial cleaning of electronic equipment, public facilities, medical and health care, business offices or food packaging surfaces.

Description

Technical Field

[0001] The present invention relates to the field of chemistry, in particular to a photosensitizer. Background Art

[0002] Photodynamic therapy (PDT) is a non-invasive treatment for cancer and bacterial infections that has attracted much attention due to its precise spatiotemporal release, low drug dependence, and minimal side effects. This method uses light to excite photosensitizers to a short-lived singlet excited state, which then travels to a triplet excited state via intersystem crossing, and then produces highly cytotoxic reactive oxygen species through type I (electron transfer) or type II (energy transfer) photochemical mechanisms, thereby achieving the purpose of killing pathogenic bacteria and cancer cells. As a major component of photodynamic therapy, photosensitizers play a vital role in enhancing the effect of photodynamic therapy and guiding treatment with fluorescence imaging. However, improving the reactive oxygen production capacity and imaging sensitivity of photosensitizers remains a huge challenge facing current photodynamic therapy.

[0003] Traditional photosensitizers often exhibit poor imaging sensitivity and low ROS generation efficiency due to the quenching properties caused by aggregation. This makes the photodynamic therapy effect strongly dependent on the environment, that is, it is difficult for these photosensitizers to simultaneously produce free radicals with low oxygen dependence (type I reactive oxygen species) and singlet oxygen with high oxygen dependence (type II reactive oxygen species), which severely limits their application in inhibiting bacterial infection, antibacterial cleaning and tumor treatment. Summary of the invention

[0004] The object of the present invention is to provide an aggregation-induced emission photosensitizer that simultaneously generates type I and type II reactive oxygen species, so as to solve the above-mentioned problems.

[0005] Another object of the present invention is to provide an application of an aggregation-induced emission photosensitizer that simultaneously generates type I and type II reactive oxygen species.

[0006] The technical problem solved by the present invention can be achieved by adopting the following technical solutions:

[0007] An aggregation-induced emission photosensitizer that simultaneously generates type I and type II reactive oxygen species, characterized in that its chemical structural formula is:

[0008]

[0009] Among them, R 1 , R 2 , R 3 and R 4 is any one of aliphatic hydrocarbon, aliphatic hydrocarbon derivative group, aromatic hydrocarbon, and aromatic hydrocarbon derivative group, and at least one of them is a positively charged substituent; R 5is any one of hydrogen, hydroxyl, dimethylamino, trimethylamino, halogen or carboxyl, wherein n=0-12; R 6 is any one of the conjugated groups.

[0010] Preferably, R 1 is hydrogen; R 2 The curve marks the substitution position, R 7 is aliphatic hydrocarbon or ethyl; X is PF 6 - or I - ; R 3 and R 4 is hydrogen; R 5 is hydrogen, wherein n=0 to 12; R 6 for The curve marks the substitution position; R 8 and R 9 It is either hydrogen or methoxy.

[0011] The preferred chemical structural formula is:

[0012]

[0013]

[0014] The applications of aggregation-induced luminescence photosensitizers that simultaneously produce type I and type II reactive oxygen species are: 1) inhibiting and treating bacterial infections, 2) inactivating microorganisms for antibacterial cleaning, and 3) killing tumors for tumor treatment.

[0015] The photosensitizer of the present invention can simultaneously generate type I and type II reactive oxygen species, effectively inhibit and treat bacterial infections, inactivate microorganisms and kill tumor tissues through photodynamic therapy. Experimental results show that the photosensitizer of the present invention exhibits a wide absorption spectrum that matches white light, and the amount of reactive oxygen species generated is much higher than that of commercial dyes, and has a better photodynamic effect. The photosensitizer of the present invention belongs to aggregation-induced luminescence photosensitizers. This type of photosensitizer reduces the thermal inactivation of excitation energy by limiting intramolecular motion (RIM), thereby generating bright fluorescence and enhanced photosensitivity in an aggregated state, which has guiding significance for the development of type I and type II photosensitizers. The present invention effectively improves the environmental limitation problem of photodynamic therapy, and is expected to replace commercial photosensitizers to achieve multiple applications in multiple occasions, such as antibacterial and anti-tumor in vivo, and antibacterial cleaning of electronic equipment, public facilities, medical and health care, business offices or food packaging surfaces. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 .H-NMR spectrum of photosensitizer I-5.

[0017] Figure 2.Comparison of the total reactive oxygen species production capacity of photosensitizer I-5 with commercial photosensitizers Rose Bengal (RB) and chlorin e6 (Ce6) under the same lighting conditions.

[0018] Figure 3 .Comparison of the type I reactive oxygen species production capacity of photosensitizer I-5 with commercial photosensitizers Rose Bengal (RB) and chlorin e6 (Ce6) under the same lighting conditions.

[0019] Figure 4 .Comparison of the hydroxyl radical generating ability of photosensitizer I-5 with commercial photosensitizers Rose Bengal (RB) and chlorin e6 (Ce6) under the same illumination conditions.

[0020] Figure 5 .Comparison of the type II reactive oxygen species production capacity of photosensitizer I-5 with commercial photosensitizers Rose Bengal (RB) and chlorin e6 (Ce6) under the same lighting conditions.

[0021] Figure 6 .Toxicity of photosensitizer I-5 to Escherichia coli or Staphylococcus aureus under dark or light conditions.

[0022] Figure 7 .Photographs of photodynamic therapy of wounds of bacterially infected mice using photosensitizer I-5 under dark or light conditions.

[0023] Figure 8 .Toxicity of photosensitizer I-5 to Hela cells under normoxic, hypoxic or anaerobic conditions.

[0024] Fig. 9 .The effect of photosensitizer I-5 on photodynamic therapy of mouse tumors under dark or light conditions.

[0025] Fig.10 .Statistical graph of the toxicity test of photosensitizer on Hela cells under normoxic, hypoxic or anaerobic conditions.

[0026] Fig.11 .A diagram showing the effect of photosensitizer on photodynamic therapy of mouse tumors. DETAILED DESCRIPTION

[0027] In order to make the technical means, creative features, objectives and effects achieved by the present invention easy to understand, the present invention is further described below with reference to specific illustrations.

[0028] About Photosensitizers

[0029] The aggregation-induced luminescence photosensitizer that simultaneously generates type I and type II reactive oxygen species is a compound represented by formula I

[0030]

[0031] In Formula I, R 1 , R 2 , R 3 and R 4 is any one of aliphatic hydrocarbon, aliphatic hydrocarbon derivative group, aromatic hydrocarbon, and aromatic hydrocarbon derivative group, and at least one of them is a positively charged substituent; R 5 is any one of hydrogen, hydroxyl, dimethylamino, trimethylamino, halogen or carboxyl, wherein n=0-12; R 6 is any one of the conjugated groups.

[0032] Preferred solution 1, R 1 is hydrogen; R 2 is a group shown in formula II (wherein the curve mark is the substitution position, the same below); R 3 and R 4 is hydrogen; R 5 is hydrogen, wherein n=0 to 12; R 6 is a group shown in formula III; in formula II, R 7 is an aliphatic hydrocarbon, X is preferably BF 4 - PF 6 - ,I - Br - , Cl - or HSO 3 - Any one of; R in Formula III 8 and R 9 Any one selected from hydrogen, methyl or methoxy.

[0033]

[0034] Preferred solution 2, R 1 is hydrogen; R 2 is a group represented by formula II, wherein R 7 is ethyl, X is PF 6 - or I - Any one of R 3 and R 4 is hydrogen; R 5 is hydrogen, wherein n=0 to 12; R 6 is a group represented by formula III, wherein R 8 and R 9 It is either hydrogen or methoxy.

[0035] Preferred solution 3, R 1 is hydrogen; R 2is a group represented by formula II, wherein R 7 is ethyl, X is PF 6 - or I - Any one of R 3 and R 4 is hydrogen; R 5 is hydrogen, wherein n=2; R 6 is a group represented by formula III, wherein R 8 and R 9 For hydrogen.

[0036] Preferred solution 4, R 1 is hydrogen; R 2 is a group represented by formula II, wherein R 7 is ethyl, X is PF 6 - ; R 3 and R 4 is hydrogen; R 5 is hydrogen, wherein n=2; R 6 is a group represented by formula III, wherein R 8 and R 9 For hydrogen. Specific embodiment 1

[0038] The aggregation-induced luminescence photosensitizer that simultaneously generates type I and type II reactive oxygen species is a compound represented by formula I-1:

[0039]

[0040] The synthesis method of the compound represented by formula I-1:

[0041]

[0042] In a 250mL single-necked flask, add Ⅱ-1 (187.3mg, 0.6mmol), 5-(4-(bis(4-methoxyphenyl)amino)phenyl)thiophene-2-carboxaldehyde (207.6mg, 0.5mmol), piperidine (1.0mL) and acetonitrile (50.0mL), and react at 90℃ for 8h under nitrogen protection. Remove the solvent by distillation under reduced pressure, dissolve the crude product in methanol, and slurry it with n-hexane to obtain 142.9mg of orange-red intermediate Ⅲ-2 with a yield of 40.3%. In a 100mL single-necked flask, add Ⅲ-2 (141.9mg, 0.2mmol), iodopropane (1.0mL), acetonitrile (20.0mL), and react at 90℃ for 10h under nitrogen protection. After removing acetonitrile under reduced pressure, add KPF 6 Saturated acetone solution (5.0 mL) was used for ion exchange for 4 h. After removing acetone by vacuum distillation, the crude product was dissolved in dichloromethane and washed with brine three times. The organic layers were combined and purified by anhydrous Na 2 SO 4The solvent was removed by drying and evaporation, and the crude product was purified by silica gel column chromatography (dichloromethane:methanol=30:1) to obtain 98.2 mg of red solid photosensitizer I-1 with a yield of 54.7%. Specific embodiment 2

[0044] The aggregation-induced luminescence photosensitizer that simultaneously generates type I and type II reactive oxygen species is a compound represented by formula I-2:

[0045]

[0046] The synthesis method of the compound shown in formula I-2:

[0047]

[0048] In a 250mL single-necked flask, add Ⅱ-1 (187.3mg, 0.6mmol), 5-(4-(bis(4-methoxyphenyl)amino)phenyl)thiophene-2-carboxaldehyde (207.6mg, 0.5mmol), piperidine (1.0mL) and acetonitrile (50.0mL), and react at 90℃ for 8h under nitrogen protection. Remove the solvent by distillation under reduced pressure, dissolve the crude product in methanol, and slurry it with n-hexane to obtain 184.8mg of orange-red intermediate Ⅲ-2, with a yield of 52.1%. In a 100mL single-necked flask, add Ⅲ-2 (141.9mg, 0.2mmol), iodoethane (1.0mL), acetonitrile (20.0mL), and react at 90℃ for 10h under nitrogen protection. After removing acetonitrile under reduced pressure, add KPF 6 Saturated acetone solution (5.0 mL) was used for ion exchange for 4 h. After removing acetone by vacuum distillation, the crude product was dissolved in dichloromethane and washed with brine three times. The organic layers were combined and purified by anhydrous Na 2 SO 4 The solvent was removed by drying and evaporation, and the crude product was purified by silica gel column chromatography (dichloromethane:methanol=30:1) to obtain 74.7 mg of red solid photosensitizer I-1 with a yield of 42.3%. Specific embodiment 3

[0050] The aggregation-induced luminescence photosensitizer that simultaneously generates type I and type II reactive oxygen species is a compound represented by formula I-3:

[0051]

[0052] The synthesis method of the compound shown in formula I-3:

[0053]

[0054] Add Ⅱ-2 (271.4 mg, 0.6 mmol), 5-(4-(diphenylamino)phenyl)thiophene-2-carboxaldehyde (177.6 mg, 0.5 mmol), piperidine (1.0 mL) and acetonitrile (50.0 mL) to a 250 mL single-necked flask and react at 90 °C for 8 h under nitrogen protection. Remove the solvent by distillation under reduced pressure, dissolve the crude product in methanol, and slurry it with n-hexane to obtain 238.4 mg of orange-red intermediate Ⅲ-3 with a yield of 60.4%. Add Ⅲ-3 (141.9 mg, 0.2 mmol), iodoethane (1.0 mL), acetonitrile (20.0 mL) to a 100 mL single-necked flask and react at 90 °C for 10 h under nitrogen protection. After removing acetonitrile under reduced pressure, add KPF 6 Saturated acetone solution (5.0 mL) was used for ion exchange for 4 h. After removing acetone by vacuum distillation, the crude product was dissolved in dichloromethane and washed with brine three times. The organic layers were combined and purified by anhydrous Na 2 SO 4 The solvent was removed by drying and evaporation, and the crude product was purified by silica gel column chromatography (dichloromethane:methanol=30:1) to obtain 103.2 mg of red solid photosensitizer I-1 with a yield of 54.6%. Specific embodiment 4

[0056] The aggregation-induced luminescence photosensitizer that simultaneously generates type I and type II reactive oxygen species is a compound represented by formula I-4:

[0057]

[0058] The type I and type II photosensitizers with aggregation-induced emission properties can also be compounds represented by formula I-5, and the nuclear magnetic hydrogen spectrum is as follows Figure 1 As shown:

[0059]

[0060] The synthesis method of the compound represented by formula I-4 and the compound represented by formula I-5:

[0061]

[0062] In a 250mL single-necked flask, Ⅱ-1 (187.3mg, 0.6mmol), 5-(4-(diphenylamino)phenyl)thiophene-2-carboxaldehyde (177.6mg, 0.5mmol), piperidine (1.0mL) and acetonitrile (50.0mL) were added, and the mixture was reacted at 90°C for 8h under nitrogen protection. The solvent was removed by distillation under reduced pressure, and the crude product was dissolved in methanol and slurried with n-hexane to obtain 186.8mg of orange-red intermediate Ⅲ-1 with a yield of 57.5%. In a 100mL single-necked flask, Ⅲ-1 (129.8mg, 0.2mmol), iodoethane (1.0mL), acetonitrile (20.0mL) were added, and the mixture was reacted at 90°C for 10h under nitrogen protection. After removing acetonitrile under reduced pressure, the crude product was purified by silica gel column chromatography (dichloromethane: methanol = 30:1) to obtain 108.1 of photosensitizer I-4 with a yield of 67.1%. Add KPF to photosensitizer I-4 (80.5 mg, 0.1 mmol) 6 Saturated acetone solution (5.0 mL) was used for ion exchange for 4 h. After removing acetone by vacuum distillation, the crude product was dissolved in dichloromethane and washed with brine three times. The organic layers were combined and purified by anhydrous Na 2 SO 4 The solvent was removed by drying and evaporation, and the crude product was purified by silica gel column chromatography (dichloromethane: methanol = 30: 1) to obtain 40.0 mg of red solid photosensitizer I-5 with a yield of 48.6%.

[0063] The total active oxygen generation capacity of the photosensitizer in the PBS system is shown in Table 2. Figure 2 :

[0064] In white light (400-650nm, 40mW cm -2 ) was used as the indicator, and the commercial photosensitizers Rose Bengal (RB) and chlorin e6 (Ce6) were used as references to test the total reactive oxygen species production capacity of 10 μM photosensitizers in PBS system. After 60 seconds of illumination, the fluorescence intensity of the indicator at 530 nm was enhanced by 86.4 times, 91.0 times, 120.7 times, 127.9 times, and 132.9 times, respectively, which was much higher than that of the commercial photosensitizers RB (32.7 times) and Ce6 (3.4 times), indicating that the photosensitizers I-1, I-2, I-3, I-4, and I-5 can quickly produce a large amount of reactive oxygen species.

[0065] The photosensitizer's ability to generate type I reactive oxygen species in the PBS system is shown in Table 1. Figure 3 :

[0066] In white light (400-650nm, 40mW cm -2) irradiation, using dihydrorhodamine 123 (DHR123) as an indicator, and commercial photosensitizers Rose Bengal (RB) and chlorin e6 (Ce6) as references, the type I reactive oxygen species generation capacity of 10 μM photosensitizer in PBS system was tested. Figure 3 As shown, in the presence of the commercial photosensitizer chlorin e6 (Ce6), the fluorescence intensity of DHR123 hardly changes with time, indicating that chlorin e6 (Ce6) does not produce type I reactive oxygen species. The generation of type I reactive oxygen species was further verified by the free radical scavenger vitamin C (Vc). Compared with photosensitizer I-5, the fluorescence intensity of DHR123 triggered by RB was lower, and the addition of vitamin C only inhibited the fluorescence to 90%, proving that RB is a type II photosensitizer. In contrast, photosensitizer I-5 triggered high fluorescence of DHR123, and the fluorescence was severely inhibited to 6% after the addition of vitamin C; photosensitizers I-1, I-2, I-3 and I-4 triggered high fluorescence of DHR123, and the fluorescence was severely inhibited to 15%-7% after the addition of vitamin C, indicating that the photosensitizer can produce type I reactive oxygen species.

[0067] The hydroxyl radical generating ability of photosensitizer in PBS system is shown in Table 2. Figure 4 :

[0068] In white light (400-650nm, 40mW cm -2 ) irradiation, with hydroxyphenylfluorescein (HPF) as an indicator, and commercial photosensitizers Rose Bengal (RB) and chlorin e6 (Ce6) as references, the hydroxyl radical generation ability of 10 μM photosensitizer in PBS system was tested. When Rose Bengal (RB) and chlorin e6 (Ce6) were added to the HPF solution, almost no fluorescence enhancement was observed, indicating that Rose Bengal (RB) and chlorin e6 (Ce6) do not generate hydroxyl radicals. When photosensitizers I-1, I-2, I-3, I-4, and I-5 were added, the fluorescence at 515 nm increased rapidly with the increase of illumination time, indicating that photosensitizers I-1, I-2, I-3, I-4, and I-5 can generate hydroxyl radicals under illumination.

[0069] The photosensitizer's ability to produce type II reactive oxygen species in the PBS system is shown in Table 2. Figure 5 :

[0070] In white light (400-650nm, 40mW cm -2) irradiation, with 9',10'-anthracene dioylbis(methylene)dihaloic acid (ABDA) as an indicator, and commercial photosensitizers Rose Bengal (RB) and chlorin e6 (Ce6) as references, the type II reactive oxygen species generation ability of 10μM photosensitizer in PBS system was tested. After 3 minutes of white light irradiation, the light absorption intensity of ABDA in the group without photosensitizer was almost unchanged, while the absorbance of Rose Bengal (RB) and chlorin e6 (Ce6) groups decreased by 19.2% and 10.2%, respectively, and the absorbance of photosensitizers I-1, I-2, I-3, I-4, and I-5 groups decreased rapidly by 50.4%, 53.7%, 60.1%, 61.8%, and 62.2%, indicating that photosensitizers I-1, I-2, I-3, I-4, and I-5 have better type II ROS generation effects than commercial photosensitizers.

[0071] Confocal fluorescence images of photosensitizer co-incubated with Escherichia coli and Staphylococcus aureus, refer to Figure 6 :

[0072] A single colony was inoculated into LB medium and cultured in a shaker (37°C, 150r / min). The bacterial solution with an optical density of 0.55 at 600nm was diluted 100 times and incubated with photosensitizers (photosensitizers I-1, I-2, I-3, I-4, I-5) for 30min on a shaker (37°C, 150r / min). After incubation, the bacteria were washed with PBS and then concentrated by centrifugation (8000r / min, 5min). Slides and coverslips were used for sample preparation, and fluorescence imaging was performed using a confocal microscope Leica TCS SP8. The results showed that after incubation with photosensitizers, Escherichia coli showed bright hollow rod fluorescence and Staphylococcus aureus showed bright round fluorescence, indicating that photosensitizers I-1, I-2, I-3, I-4, and I-5 can quickly bind to bacteria through electrostatic interactions, thereby achieving fluorescence imaging-guided photodynamic therapy.

[0073] The toxicity of photosensitizers to Escherichia coli or Staphylococcus aureus is shown in Table 1. Figure 7 :

[0074] The plate colony counting method was used to evaluate the antibacterial effect. First, 30 μL of the diluted bacterial PBS solution was evenly spread on the LB solid culture medium, and then the culture dish was inverted in a bacterial incubator at 37°C. After 24 hours, the plate was photographed and counted. The experiment set up PBS group, PBS+Light group, photosensitizer I group and photosensitizer I+Light group, with 3 parallel controls in each group. Different photosensitizer concentrations were set, which were 2.5 μM, 5 μM and 10 μM respectively. When photosensitizers I-1, I-2, I-3, I-4, and I-5 were added respectively and cultured under dark conditions, both the Escherichia coli and Staphylococcus aureus groups showed a large number of colonies, indicating that photosensitizers I-1, I-2, I-3, I-4, and I-5 have excellent biocompatibility. In contrast, after incubation with photosensitizers I-1, I-2, I-3, I-4, and I-5 and then irradiated with light, the colony counts of Escherichia coli and Staphylococcus aureus decreased sharply in a concentration-dependent manner, indicating that photosensitizers I-1, I-2, I-3, I-4, and I-5 can effectively kill bacteria under light irradiation. Figure 6 As shown, after incubation with 5 μM photosensitizer I-5, the survival rate of E. coli dropped to 2.26% and the survival rate of Staphylococcus aureus dropped to 1.64%, indicating that low concentrations of photosensitizer I-5 were sufficient to effectively kill the bacteria.

[0075] The situation of photosensitizer in photodynamic therapy of wounds of bacterially infected mice, refer to Figure 8 :

[0076] Newly purchased Balb / c male mice were raised to a body weight of 20±5g and divided into three groups (4 mice in each group): PBS group, photosensitizer I group, and photosensitizer I+Light group. On day 0, all mice were anesthetized, the back hair was removed with a razor and depilatory cream, and the exposed skin was cleaned and disinfected with 75% alcohol. On day 1, rat skin infection with Escherichia coli was established: (1) After anesthetizing the mice, two circular wounds with a diameter of 8 mm were drilled on the back of the mice with a skin biopsy punch; (2) Escherichia coli suspension (1×10 9 CFU mL -1 , 20 μL), and covered the wound with 3M transparent dressing for 2 hours; (3) After infection, 60 μL of PBS and 10 μM photosensitizer (photosensitizers I-1, I-2, I-3, I-4, I-5) solution were added to the wound and incubated for 15 minutes; at the end of treatment, the mice were raised normally; (4) The photosensitizer I+Light group was illuminated for 15 minutes at 40 mW / cm 2 On days 1, 3, 5, 8, 10, and 12, the wounds were photographed, their diameters were measured and recorded, and the weights of the mice were also recorded. Figure 7This is the treatment situation of photosensitizer I-5. After 12 days of treatment, the wounds in the control group and photosensitizer I-5 group did not disappear, while the wounds in the photosensitizer + Light group were mostly repaired, indicating that photosensitizers I-1, I-2, I-3, I-4, and I-5 accelerated the wound healing process and reduced bacterial infection of the wound through photodynamic therapy.

[0077] Confocal fluorescence images of HeLa cells co-incubated with photosensitizer, see Fig. 9 :

[0078] After HeLa cells were inoculated into a glass-bottomed culture dish (Ф20 mm, NEST), they were cultured for 12 hours. After incubation with photosensitizers (photosensitizers I-1, I-2, I-3, I-4, and I-5) for 1 hour, the solution was aspirated, the cells were washed once with PBS, fresh culture medium was added, and fluorescence imaging was performed using a confocal microscope Leica TCS SP8. The results showed that bright red fluorescence was observed in HeLa cells after incubation with photosensitizers for 1 hour, indicating that photosensitizers I-1, I-2, I-3, I-4, and I-5 have good cell penetration and imaging capabilities.

[0079] The toxicity of photosensitizers to Hela cells under normoxic, hypoxic or anaerobic conditions is shown in Table 1. Fig.10 :

[0080] The viability of HeLa cells was assessed by the methylthiazolyldiphenyltetrazolium bromide (MTT) assay. First, cells were plated at 8 × 10 3 The cells were seeded into 96-well plates at a density of 10 cells / mL and cultured for 8 h. Then, the cells were incubated with culture medium containing different concentrations of photosensitizers I-1, I-2, I-3, I-4, and I-5 (15, 10, 5, 2.5, 1, and 0 μM) for 4 h, and then the photosensitizer I solution was replaced with fresh DMEM. Illumination (40 mW cm -2 ) for 15 min or incubate in dark for 12 h. Add 10 μL MTT solution (5 mg mL -1 ), and continue to culture for 4h. After removing the supernatant, 100μL DMSO was added to each well to dissolve the crystals. The absorbance of MTT at 570nm was recorded by a microplate reader (BioTek Synergy H4) to obtain cell viability, and the cells incubated only with culture medium in each group were designated as cells with 100% cell viability. For hypoxic or anaerobic tests, the culture plates were placed in 8% or 1% hypoxic culture bags for 2h before illumination, and illumination operations were performed in hypoxic bags, and the rest of the operations were consistent. For dark toxicity tests, the illumination process was omitted, and the rest of the operations were consistent with phototoxicity tests. Hela cells treated with photosensitizers I-1, I-2, I-3, I-4, and I-5 were exposed to normoxia (21% O 2 ), hypoxia (8% O 2) and anaerobic (1% O 2 ) were all lower than 4.57%, 6.94% and 50.78%, respectively. This indicates that photosensitizers I-1, I-2, I-3, I-4 and I-5 have significant Hela cell killing effects under hypoxia. Among them, Hela cells treated with photosensitizer I-5 had a significant killing effect under normoxia (21% O 2 ), hypoxia (8% O 2 ) and anaerobic (1% O 2 ) states were 2.60%, 4.34% and 41.28% respectively.

[0081] The results of photodynamic therapy of mouse tumors by photosensitizers are shown in Figure 2. Fig.11 :

[0082] 3×10 6 HeLa cells were injected into the right back of 6-week-old female nude mice. When the tumor size reached about 200 mm 3 The mice were randomly divided into three groups (n=5 / group): PBS group, photosensitizer I group and photosensitizer I+Light group (white light 40 mW cm -2 , 15min, the photosensitizer dose is 10mg / kg). The mice were injected through the tail vein every other day, and the volume of the mice was measured and recorded. According to the equation V = ab 2 ×1 / 2 to determine the volume of the tumor, where a and b are the longest and shortest diameters of the tumor, respectively. After 9 days of treatment, all mice were euthanized, and the tumors were collected and imaged. The tumor size decreased in the order of PBS group > photosensitizer I group > photosensitizer I + Light group. After 9 days, the average tumor volume of the control group was close to 2000mm 3 , while the tumor growth rate in the photosensitizer I-5+Light group was significantly slowed down, indicating that photosensitizers I-1, I-2, I-3, I-4, and I-5 have excellent anti-tumor ability.

[0083] Photodynamic antibacterial effect of photosensitizer on surface

[0084] 20 μL of photosensitizer I-5 (10 μM) was evenly coated on the slide, and after the surface was completely dry, 10 5 Staphylococcus aureus was applied to the coating surface. The slides were kept in the dark until the slides were visibly dry. The experiment was divided into: control group + light group, control dark group, photosensitizer I + light group and photosensitizer I dark group (n = 3 / group). The control group was operated in the same way except that the photosensitizer was not applied. After illumination (white light 40mW cm -2, 15min), and placed in the dark together with the dark group for 24h, and then sterile cotton swabs were used to remove bacteria from the surface, and then placed on LB solid agar for continuous dilution and incubation at 37℃ for 24h, and the survival rate of bacteria was determined by calculating the number of colony forming units per milliliter (cfu / mL). The experimental results showed that the number of colony forming units on the surface of the control group + light group was 6.9cfu / mL, and the number of colony forming units on the surface of the photosensitizer (I-1, I-2, I-3, I-4, I-5) + light group was 1.0cfu / mL, 0.9cfu / mL, 0.6cfu / mL, 0.5cfu / mL, 0.2cfu / mL, and the bacterial survival rates were 13.5%, 12.2%, 8.1%, 6.8%, 3.0%, respectively. Therefore, photosensitizers I-1, I-2, I-3, I-4, and I-5 have good photodynamic antibacterial effects on the surface.

[0085] The above shows and describes the basic principles and main features of the present invention and the advantages of the present invention. It should be understood by those skilled in the art that the present invention is not limited to the above embodiments. The above embodiments and descriptions are only for explaining the principles of the present invention. Without departing from the spirit and scope of the present invention, the present invention may have various changes and improvements, which fall within the scope of the present invention to be protected. The scope of protection of the present invention is defined by the attached claims and their equivalents.

Claims

1. An aggregation-induced emission photosensitizer that simultaneously produces type I and type II reactive oxygen species, characterized in that: Its chemical structure is:

2. A method for preparing an aggregation-induced emission photosensitizer that simultaneously generates type I and type II reactive oxygen species, characterized in that: Will 5-(4-(bis(4-methoxyphenyl)amino)phenyl)thiophene-2-carboxaldehyde, piperidine and acetonitrile are mixed and reacted at 90°C under nitrogen protection; the solvent is distilled off under reduced pressure, the crude product is dissolved in methanol, and slurried with n-hexane to obtain intermediate III-2; III-2, iodopropane and acetonitrile are mixed and reacted at 90°C under nitrogen protection; after removing acetonitrile under reduced pressure, a saturated acetone solution of KPF6 is added for ion exchange; after removing acetone by distillation under reduced pressure, the crude product is dissolved in dichloromethane, washed with brine, the organic layers are combined, dried over anhydrous Na2SO4 and evaporated to remove the solvent, and then the crude product is purified by silica gel column chromatography to obtain 3. A method for preparing an aggregation-induced emission photosensitizer that simultaneously generates type I and type II reactive oxygen species, characterized in that: Will 5-(4-(bis(4-methoxyphenyl)amino)phenyl)thiophene-2-carboxaldehyde, piperidine and acetonitrile are reacted at 90°C under nitrogen protection; the solvent is distilled off under reduced pressure, the crude product is dissolved in methanol, and slurried with n-hexane to obtain intermediate III-2; III-2, iodoethane and acetonitrile are mixed and reacted at 90°C under nitrogen protection; after the acetonitrile is removed under reduced pressure, a saturated acetone solution of KPF6 is added for ion exchange; after the acetone is distilled off under reduced pressure, the crude product is dissolved in dichloromethane, washed with brine, the organic layers are combined, dried over anhydrous Na2SO4 and evaporated to remove the solvent, and then the crude product is purified by silica gel column chromatography to obtain 4. A method for preparing an aggregation-induced emission photosensitizer that simultaneously generates type I and type II reactive oxygen species, characterized in that: Will 5-(4-(diphenylamino)phenyl)thiophene-2-carboxaldehyde, piperidine and acetonitrile were mixed and reacted at 90°C under nitrogen protection; the solvent was removed by distillation under reduced pressure, the crude product was dissolved in methanol, and slurried with n-hexane to obtain III-1; III-1, iodoethane and acetonitrile were mixed and reacted at 90°C under nitrogen protection; after removing acetonitrile under reduced pressure, the crude product was purified by silica gel column chromatography to obtain 5. A method for preparing an aggregation-induced emission photosensitizer that simultaneously generates type I and type II reactive oxygen species, characterized in that: exist A saturated acetone solution of KPF6 was added to the mixture for ion exchange; after removing the acetone by distillation under reduced pressure, the crude product was dissolved in dichloromethane, washed with brine, the organic layers were combined, dried over anhydrous Na2SO4 and evaporated to remove the solvent, and then the crude product was purified by silica gel column chromatography to obtain 6. The use of the aggregation-induced emission photosensitizer that simultaneously generates type I and type II reactive oxygen species according to claim 1, characterized in that: Application in the preparation of photosensitizers for photodynamic therapy.

7. The use of the aggregation-induced emission photosensitizer that simultaneously generates type I and type II reactive oxygen species according to claim 1, characterized in that: Application in the preparation of surface antibacterial cleaning agents or surface antibacterial cleaning agent additives.

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