Near-infrared absorption supramolecular cationic free radical as well as preparation method and application thereof

Abstract

The invention discloses a near-infrared absorption supramolecular cation free radical as well as a preparation method and application thereof, and belongs to the technical field of photo-thermal therapy. The supramolecular cation free radical is obtained by taking a supramolecular host compound as a stabilizer, forming a compound by supramolecular self-assembly of the supramolecular host compound and a guest molecule compound in an aqueous solution, and then oxidizing to lose an electron. Compared with an original-guest cation free radical (the structural formula is shown in the formula II), the supramolecular cation free radical has better water solubility, longer free radical life, stronger near-infrared absorption capability, more excellent photothermal conversion capability and stronger capability of photothermally killing pathogenic bacteria and tumor cells.

Description

Technical Field

[0001] The present invention belongs to the technical field of photothermal therapy, and specifically relates to a supramolecular cation free radical with near-infrared absorption and the preparation and application thereof. Background Art

[0002] Organic free radicals have unique optical, electrical, magnetic and redox properties and can be used in the fields of spin probes, reaction catalysis, magnetic resonance imaging, fluorescence imaging, photothermal therapy, etc. However, most organic free radicals have poor stability and short lifetime, which seriously limits their wide application. In order to extend the lifetime of free radicals and improve the stability of free radicals, conventional methods include: 1) covalently introducing π-conjugated groups, electron-donating / electron-withdrawing groups or large steric protective groups into the free radical center; 2) using non-covalent supramolecular interactions to protect free radicals. Covalent modification methods often require complex molecular design and organic synthesis. Therefore, in recent years, non-covalent methods for stabilizing free radicals have attracted more and more attention from researchers. In addition, supramolecular free radicals also have the advantages of good water solubility and stimulus responsiveness, and have broad application prospects in the field of biological diagnosis and treatment.

[0003] There are already literature reports in the prior art that cationic free radicals can be stabilized by supramolecular interactions. For example, Chinese invention patent CN117024435A discloses a near-infrared luminescent free radical cationic compound and its preparation and application. However, the absorption spectrum of the cationic free radical of the invention does not reach the near-infrared region (maximum absorption wavelength 590-630nm), and its application in photothermal therapy has certain limitations. Summary of the invention

[0004] In view of the problems existing in the prior art, the present invention discloses a supramolecular cationic free radical with near-infrared absorption and its preparation and application, which can produce absorption in the near-infrared region and has a photothermal sterilization effect.

[0005] As one of the objects of the invention, the present invention provides a supramolecular cation free radical with near-infrared absorption, the structural formula of the supramolecular cation free radical is shown in Formula III, wherein the macrocyclic molecule is one or more of cucurbituril, cyclodextrin, calixarene and pillararene.

[0006] The near-infrared absorbing supramolecular cation free radical can produce absorption at 500-1100nm. Compared with the original guest cation free radical, the supramolecular cation free radical has better water solubility, longer free radical life, stronger near-infrared absorption ability, better photothermal conversion ability, and advantages such as sterilization and tumor killing.

[0007]

[0008] Wherein, X is methylene, carbonyl, S, O, NH; R 1 and R 2 are independently substituted or unsubstituted alkyl, alkyloxy, primary / secondary / tertiary amine.

[0009] Another object of the present invention is to provide a method for preparing the supramolecular cationic radical, which has the advantages of no need for heating, no need for adding volatile organic solvents, rapid reaction, and no need for subsequent separation.

[0010] Preferably, the synthetic route of the preparation method is:

[0011]

[0012] It can be seen from the above reaction equation that the supramolecular cation free radical provided by the present invention uses the supramolecular host compound as a stabilizer, and is obtained by supramolecular self-assembly with the guest molecule in an aqueous solution, and the guest molecule passes through the macrocyclic structure of the supramolecular host compound to form a complex, and then is oxidized to lose an electron.

[0013] Preferably, the supramolecular host compound is one or more of cucurbituril, cyclodextrin, calixarene and pillararene.

[0014] Preferably, the oxidant may be oxygen, an atmosphere containing oxygen (such as air), a peroxide (such as H 2 O 2 ), ammonium persulfate, free radical initiator (such as: I2959), hydrogen peroxide + ferrous ion, hydrogen peroxide + catalase, ferric chloride, silver nitrate, one or more of iodine.

[0015] Another object of the present invention is to provide the application of the supramolecular cationic free radical in photothermal therapy. The photothermal therapy refers to the application of photothermal killing of pathogens (including Gram-positive bacteria and Gram-negative bacteria) and tumor cells (including cervical cancer cells and breast cancer cells), which has the advantages of low dark toxicity, good killing effect, and wide application range.

[0016] Compared with the prior art, the present invention has the following technical effects:

[0017] (1) The method for preparing the near-infrared absorbing supramolecular cationic free radicals adopted by the present invention has the advantages of no need for heating, no need for adding volatile organic solvents, rapid reaction, no need for subsequent separation, etc. The process is simple and easy to control, the conditions are mild, safe and pollution-free.

[0018] (2) Compared with the original guest cation radical (structural formula see Formula II), the supramolecular cation radical provided by the present invention has better water solubility, longer free radical lifetime, stronger near-infrared absorption ability, better photothermal conversion ability and stronger photothermal killing ability of pathogens and tumor cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 The absorption spectra and actual solution images of DAF and the host-guest complex DAF@CB[8] after oxidation in Example 1 of the present invention.

[0020] Figure 2 The absorption spectrum and actual solution image of DAT and the host-guest complex DAT@CB[8] after oxidation in Example 2 of the present invention.

[0021] Figure 3 The absorption spectra and actual solution images of DAFO and the host-guest complex DAFO@CB[8] after oxidation in Example 3 of the present invention.

[0022] Figure 4 DAF in Example 7 of the present invention ·+ @CB[8]、DAT ·+ @CB[8] and DAFO ·+ Solid-state electron paramagnetic resonance spectrum of @CB[8].

[0023] Figure 5 The photothermal conversion effect of three supramolecular cationic radical aqueous solutions in Example 8 of the present invention is characterized as follows: (A), (B), and (C) are DAF ·+ @CB[8]、DAT ·+ @CB[8] and DAFO ·+ @CB[8] aqueous solution was exposed to 808 nm laser (intensity 0.5 W cm -2 ) irradiation, and the original guest free radical aqueous solutions after oxidation of DAF, DAT and DAFO provided in Comparative Examples 1 to 3 were used as control groups, and pure water was used as a blank control group.

[0024] Figure 6 The photothermal stability of three supramolecular cation radical aqueous solutions in Example 8 of the present invention is characterized as follows: (A), (B), and (C) are DAF ·+ @CB[8]、DAT ·+ @CB[8] and DAFO ·+ @CB[8] aqueous solution was exposed to 808 nm laser (intensity 0.5 W cm -2 ) Photothermal stability test under irradiation.

[0025] Figure 7a and Figure 7b DAF in Example 9 of the present invention ·+ @CB[8] at 808 nm laser (intensity 0.5 W cm -2 ) Experimental results of photothermal killing of Staphylococcus aureus under irradiation.

[0026] Figure 8a and Figure 8b DAF in Example 9 of the present invention ·+ @CB[8] at 808 nm laser (intensity 0.5 W cm -2 ) Experimental results of photothermal killing of Escherichia coli under irradiation.

[0027] Fig. 9 DAF in Example 10 of the present invention ·+ @CB[8] is darkly toxic to MDA-MB-231 and HeLa cells.

[0028] Fig.10 DAF in Example 11 of the present invention ·+ @CB[8] at 808 nm laser (intensity 0.5 W cm -2 ) Experimental results of photothermal killing of MDA-MB-231 cells under irradiation.

[0029] Fig.11 DAF in Example 11 of the present invention ·+ @CB[8] at 808 nm laser (intensity 0.5 W cm -2 ) Experimental results of photothermal killing of HeLa cells under irradiation. DETAILED DESCRIPTION

[0030] The technical solution of the present invention is described clearly and completely below in conjunction with specific embodiments and drawings. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0031] The present invention provides a supramolecular cation free radical with near-infrared absorption, wherein the structural formula of the supramolecular cation free radical contains a macrocyclic molecule; the macrocyclic molecule is a combination of one or two or more of cucurbituril, cyclodextrin, calixarene and pillararene. The supramolecular cation free radical provided by the present invention is obtained by using a supramolecular host compound as a stabilizer, forming a complex with a guest molecule in an aqueous solution through supramolecular self-assembly, and then being oxidized to lose an electron. Compared with the original guest cation free radical (structural formula see formula II), the supramolecular cation free radical has better water solubility, longer free radical life, stronger near-infrared absorption ability, more excellent photothermal conversion ability and stronger photothermal killing ability of pathogens and tumor cells.

[0032] The structural formula of the original guest cation radical is shown in Formula II:

[0033]

[0034] Wherein, X is one of methylene, carbonyl, S, O, and NH; R 1 and R 2 Each of them is independently any one of a substituted or unsubstituted alkyl group, an alkyloxy group, and a primary / secondary / tertiary amine group.

[0035] The structural formula of the supramolecular cation radical is shown in Formula III:

[0036]

[0037] Wherein, X is one of methylene, carbonyl, S, O, and NH; R 1 and R 2 Each of them is independently any one of a substituted or unsubstituted alkyl group, an alkyloxy group, and a primary / secondary / tertiary amine group.

[0038] The present invention also provides a method for preparing the supramolecular cationic free radical as described above, which specifically comprises: first preparing the supramolecular host compound and the guest molecule compound into a self-assembled complex, and then adding an oxidant to oxidize the complex to lose an electron.

[0039] In some specific embodiments, the supramolecular host compound is one or a combination of two or more of cucurbituril, cyclodextrin, calixarene and pillararene.

[0040] In some specific embodiments, the structure of the guest molecule compound is as shown in Formula I, wherein X is methylene, carbonyl, S, O, NH; R 1 and R 2 It is any of a substituted or unsubstituted alkyl group, an alkyloxy group, and a primary / secondary / tertiary amine group.

[0041]

[0042] In some specific embodiments, the oxidant can be one or a combination of two or more of oxygen, an atmosphere containing oxygen, peroxide, ammonium persulfate, a free radical initiator, hydrogen peroxide+ferrous ions, hydrogen peroxide+catalase, ferric chloride, silver nitrate, and iodine.

[0043] Preferably, the atmosphere containing oxygen is air.

[0044] Preferably, the peroxide is H 2 O 2 .

[0045] Preferably, the free radical initiator is I2959.

[0046] The present invention also provides the use of the supramolecular cationic free radical in photothermal killing of pathogenic bacteria.

[0047] In some specific embodiments, the application includes adding supramolecular cationic radicals into an aqueous solution containing pathogenic bacteria to achieve the purpose of antibacterial.

[0048] Preferably, the pathogen is a Gram-positive or Gram-negative bacterium.

[0049] The present invention also provides the use of the supramolecular cation free radical in photothermal killing of tumor cells.

[0050] In some specific embodiments, the application comprises adding supramolecular cationic radicals to a culture medium containing tumor cells.

[0051] Preferably, the tumor cells are cervical cancer cells and breast cancer cells.

[0052] Example 1

[0053] This example provides a representative method for preparing supramolecular cation radicals and their absorption spectrum characterization.

[0054] In the preparation method, cucurbitacin[8]uril (CB[8]) is used as the supramolecular host compound, 2,7-diaminofluorene (DAF) is used as the guest compound, and hydrogen peroxide (H 2 O 2 )+ferrous chloride (FeCl 2 ) as an oxidant. The specific preparation method is as follows:

[0055] (1) Weigh 1.33 mg of CB[8] and dissolve it in 10 mL of water to a final concentration of 100 μM to obtain a CB[8] solution;

[0056] (2) Adding DAF ethanol solution to the CB[8] solution to make the final concentration of 100 μM, the supramolecular complex DAF@CB[8] solution was prepared;

[0057] (3) Add H 2 O 2 and FeCl 2 The final concentration of the solution was 100 μM. After shaking, let it stand for 10 minutes to obtain the supramolecular cationic radical DAF. ·+ @CB[8].

[0058] Example 2

[0059] This example provides a representative method for preparing supramolecular cation radicals and their absorption spectrum characterization.

[0060] In the preparation method, cucurbitacin[8]uril (CB[8]) is used as the supramolecular host compound, dibenzo[b, d]thiophene-3,7-diamine (DAT) is used as the guest compound, and hydrogen peroxide (H 2 O 2 )+ferrous chloride (FeCl 2 ) as an oxidant. The specific preparation method is as follows:

[0061] (1) Weigh 1.33 mg of CB[8] and dissolve it in 10 mL of water to a final concentration of 100 μM to obtain a CB[8] solution;

[0062] (2) Adding DAT ethanol solution to the CB[8] solution to make the final concentration of DAT ethanol solution 100 μM, the supramolecular complex DAT@CB[8] solution was prepared;

[0063] (3) Add H to the supramolecular complex DAT@CB[8] solution. 2 O 2 and FeCl 2 The final concentration of the solution was 100 μM. After shaking, let it stand for 10 minutes to obtain the supramolecular cation radical DAT. ·+ @CB[8].

[0064] Example 3

[0065] This example provides a representative method for preparing supramolecular cation radicals and their absorption spectrum characterization.

[0066] In the preparation method, cucurbitacin[8]uril (CB[8]) is used as the supramolecular host compound, 2,7-diamino-9H-fluorene-9-one (DAFO) is used as the guest compound, and hydrogen peroxide (H 2 O 2)+ferrous chloride (FeCl 2 ) as an oxidant. The specific preparation method is as follows:

[0067] (1) Weigh 1.33 mg of CB[8] and dissolve it in 10 mL of water to a final concentration of 100 μM;

[0068] (2) adding DAFO ethanol solution to a final concentration of 100 μM to prepare the supramolecular complex DAFO@CB[8] solution;

[0069] (3) Add H to the supramolecular complex DAFO@CB[8] solution. 2 O 2 and FeCl 2 The final concentration of the solution was 100 μM. After shaking, let it stand for 10 minutes to obtain the supramolecular cationic free radical DAFO. ·+ @CB[8].

[0070] Example 4

[0071] The only difference between this embodiment and embodiment 1 is that in step (2), the supramolecular host compound is cyclodextrin with a final concentration of 100 μM, and the other steps are the same.

[0072] Example 5

[0073] The difference between this example and Example 1 is that in step (2), the supramolecular host compound is calixarene (4-sulfonylcalix[6]arene, Shanghai Aladdin Biochemical Technology Co., Ltd., catalog number S161016), the final concentration is 100 μM, and the other steps are the same.

[0074] Example 6

[0075] The difference between this embodiment and embodiment 1 is that in step (2), the supramolecular host compound is pillar aromatic hydrocarbon (dimethoxy pillar [5] aromatic hydrocarbon, Shanghai Aladdin Biochemical Technology Co., Ltd., catalog number D154968), the final concentration is 100 μM, and the other steps are the same.

[0076] Comparative Example 1

[0077] The difference between this comparative example and Example 1 is that step (1) is not included, that is, the supramolecular host compound cucurbitacin[8]uril (CB[8]) is not included. The other steps are the same to prepare the original guest free radical.

[0078] For the aqueous solution of the original guest radical cation radical prepared in Example 1 and Comparative Example 1, the absorption spectrum in the range of 300-1200 nm was tested using a UV-visible-near infrared spectrophotometer (Agilent, Cary 5000). Figure 1As shown, if there is no supramolecular host compound CB[8] in the system (Comparative Example 1), the free radicals (DAF ·+ ) is very unstable, so there is no obvious near-infrared absorption peak, and the solution is light brown; when CB[8] is present in the system (Example 1), CB[8] can stabilize the guest molecule free radical, thus showing an obvious near-infrared absorption peak, and the solution is blue. DAF ·+ The maximum absorption peaks of @CB[8] are 607nm and 822nm.

[0079] Comparative Example 2

[0080] The difference between this comparative example and Example 2 is that step (1) is not included, that is, the supramolecular host compound cucurbitacin[8]uril (CB[8]) is not included. The other steps are the same to prepare the original guest free radical.

[0081] For the supramolecular cationic radical aqueous solutions prepared in Example 2 and Comparative Example 2, the absorption spectra in the range of 300-1200 nm were measured using a UV-visible-near infrared spectrophotometer (Agilent, Cary 5000). Figure 2 As shown, if there is no supramolecular host compound CB[8] in the system (Comparative Example 2), the free radicals (DAT ·+ ) is very unstable, so there is no obvious near-infrared absorption peak, and the solution is light brown; when CB[8] is present in the system (Example 2), CB[8] can stabilize the guest molecule free radical, thus showing an obvious near-infrared absorption peak, and the solution is blue. DAT ·+ The maximum absorption peaks of @CB[8] are 626nm and 859nm.

[0082] Comparative Example 3

[0083] The difference between this comparative example and Example 3 is that step (1) is not included, that is, the supramolecular host compound cucurbitacin[8]uril (CB[8]) is not included. The other steps are the same to prepare the original guest free radical.

[0084] For the supramolecular cationic radical aqueous solutions prepared in Example 3 and Comparative Example 3, the absorption spectra in the range of 300-1200 nm were measured using a UV-visible-near infrared spectrophotometer (Agilent, Cary 5000). Figure 3 As shown, if there is no supramolecular host compound CB[8] in the system (Comparative Example 3), the free radicals (DAFO ·+) is very unstable, so there is no obvious near-infrared absorption peak, and the solution is light brown; when CB[8] is present in the system (Example 3), CB[8] can stabilize the guest molecule free radical, thus showing an obvious near-infrared absorption peak, and the solution is blue. ·+ The maximum absorption peaks of @CB[8] are 626nm and 851nm.

[0085] The supramolecular cation radicals and original guest cation radicals prepared based on the above embodiments and / or comparative examples were subjected to structural characterization, photothermal performance characterization, and photothermal sterilization performance, cell killing effect and tumor cell killing effect tests.

[0086] Example 7

[0087] This example provides an electron paramagnetic resonance characterization method for supramolecular cation radicals.

[0088] The supramolecular cationic radical solutions (10 mL, 100 μM) prepared in Examples 1 to 3 were respectively dehydrated using a freeze dryer to prepare solid powders, which were then transferred into a quartz tube and subsequently tested for signals in the range of 3400-3600 G using an electron paramagnetic resonance spectrometer (Bruker, E500).

[0089] The results are as follows Figure 4 As shown, DAF ·+ @CB[8]、DAT ·+ @CB[8] and DAFO ·+ The g-factors of @CB[8] are 2.0035, 2.0038, and 2.0049, respectively, which are very close to the theoretical g-factor value of free electrons (2.0023), confirming that all three complexes have free radical structures.

[0090] At the same time, the g-factor of Examples 2 to 8 is similar to the DAF obtained in Example 4. ·+ @CB[8]、DAT ·+ @CB[8] and DAFO ·+ The g-factor of the supramolecular cation radical of @CB[8] is close and will not be elaborated here.

[0091] Example 8

[0092] This example provides a method for studying the photothermal properties of supramolecular cation radicals.

[0093] 200 μL of the supramolecular cationic radical solution (concentration 100 μM) prepared in Example 1 to Example 3 and the original guest radical (concentration 100 μM) prepared in Comparative Example 1 to Comparative Example 3 were added to a 0.6 mL centrifuge tube.

[0094] The output optical power of the 808 nm laser (Changchun Radium Optoelectronics, LR-MFJ-808 / 5000 mW) was adjusted to 0.5 W cm using a laser power meter (Changchun Radium Optoelectronics, VLP-2000-10 W). -2 Then, the laser was used to irradiate the aqueous solution of supramolecular cation radicals, and a thermal imager (FLIR E8-XT, USA) was used to record the temperature change of the solution. Figure 5 DAF ·+ @CB[8]、DAT ·+ @CB[8] and DAFO ·+ @CB[8] The aqueous solutions of three supramolecular cation radicals were illuminated by 808 nm laser (intensity 0.5 W cm -2 ) After irradiation for 10 min, the temperatures increased by 29.1°C, 26.3°C and 24.1°C respectively; for DAF without CB[8] protection ·+ , DAT ·+ and DAFO ·+ Under the same experimental conditions, the temperature only increased by 5.1°C, 1.3°C and 4.6°C. Subsequently, the photostability of supramolecular cation radicals after laser irradiation was tested. Figure 6 As shown, after 5 cycles of laser on-off, there is no obvious difference in the photothermal conversion curve.

[0095] The above results show that compared with the guest molecular radicals themselves, the three supramolecular cation radicals DAF ·+ @CB[8]、DAT ·+ @CB[8] and DAFO ·+ @CB[8] all have excellent photothermal conversion properties and good photothermal stability.

[0096] Meanwhile, the photothermal conversion properties and photothermal stability of Examples 2 to 8 are similar and will not be described in detail here.

[0097] Example 9

[0098] This embodiment provides a method for testing the photothermal sterilization properties of supramolecular cationic radicals.

[0099] For the supramolecular cationic radical DAF prepared in Example 1 ·+ @CB[8] tested its photothermal sterilization properties. The specific steps are as follows:

[0100] (1) Cultivating Staphylococcus aureus or Escherichia coli in LB medium to the logarithmic phase;

[0101] (2) Centrifuge the bacterial solution and then resuspend the bacteria in PBS buffer solution. Repeat the centrifugation-resuspending operation three times. Finally, resuspend the bacteria in PBS and adjust its OD 600nm is 1.0;

[0102] (3) Take 1 mL of DAF prepared in Example 1 ·+ @CB[8] aqueous solution, add 50 μL 20× PBS to make the final PBS concentration 1×;

[0103] (4) Take 360 ​​μL of the above DAF ·+ @CB[8] PBS solution, add 40 μL OD 600nm = 1.0 bacterial suspension, the final concentration of bacteria is about 0.1OD, DAF ·+ The final concentration of @CB[8] was approximately 80 μM;

[0104] (5) Using 808nm laser (0.5W cm -2 ) Irradiate the bacterial solution for 15 min, then dilute and plate.

[0105] DAF ·+ The results of the bactericidal experiments of @CB[8] on Gram-positive bacteria S. aureus and Gram-negative bacteria E. coli are as follows: Figure 7a , Figure 7b and Figure 8a , Figure 8b DAF ·+ @CB[8] showed excellent photothermal sterilization effect. Under the above experimental conditions, DAF ·+ @CB[8] has a bactericidal rate of 99.999906% against S. aureus and 99.99989% against E. coli.

[0106] At the same time, Examples 2 to 8 also have similar photothermal sterilization effects as the supramolecular free radicals of Example 1, which will not be described in detail here.

[0107] Example 10

[0108] This example provides a method for testing the in vitro biocompatibility of supramolecular cation radicals.

[0109] For the supramolecular cationic radical DAF prepared in Example 1 ·+ @CB[8], tested its biocompatibility with cells without applying light, the specific steps are as follows:

[0110] (1) Sample preparation: 10 mL DAF ·+ @CB[8] solution (100 μM) was freeze-dried to remove water, and then 5 mL of complete culture medium, i.e., DMEM high-glucose medium (Thermo Fisher Scientific, USA, Catalog No. 11965092) + 10% fetal bovine serum (PAN, Germany, Catalog No. ST30-3302) + 1% penicillin-streptomycin (Shanghai Sangon Biotechnology Co., Ltd., Catalog No. E607011) was added, and pipetting and sonication were performed to make the final concentration of 200 μM;

[0111] (2) Human cervical cancer cells HeLa and human breast cancer cells MDA-MB-231 were passaged into 96-well plates at a cell density of 8000 cells / well and then transferred to a cell culture incubator for 24 h;

[0112] (3) The cell culture medium in the 96-well plate was replaced with 5, 10, 25, 50, 100, 150, and 200 μM DAF. ·+ @CB[8] complete medium, 6 parallel wells for each concentration;

[0113] (4) After culturing for 24 h, the cell viability was tested using the MTT method.

[0114] DAF ·+ @CB[8] The biocompatibility test results of HeLa and MDA-MB-231 are as follows Fig. 9 As shown. Without light, DAF concentrations below 150 μM ·+ @CB[8] did not show obvious cell killing effect. This result shows that under the above experimental conditions, DAF ·+ @CB[8] has good biocompatibility.

[0115] At the same time, Examples 2 to 8 also have similar cell killing effects as the supramolecular free radicals in Example 1, indicating that the supramolecular free radicals prepared in Examples 2 to 8 also have the same biocompatibility, which will not be described in detail here.

[0116] Embodiment 11

[0117] This example provides a method for testing the ability of supramolecular cation radicals to kill tumor cells by photothermal therapy.

[0118] For the supramolecular cationic radical DAF prepared in Example 1 ·+ @CB[8], tested its photothermal killing properties of tumor cells, the specific steps are as follows:

[0119] (1) Human cervical cancer cells HeLa and human breast cancer cells MDA-MB-231 were passaged into 96-well plates at a cell density of 8000 cells / well and then transferred to a cell culture incubator for 24 h;

[0120] (2) Replace the cell culture medium in the 96-well plate with one containing 80 μM DAF ·+ @CB[8] or DAF ·+ Complete medium with 3 parallel wells for each concentration;

[0121] (3) Using 808nm laser (light intensity 0.5W cm -2 ) irradiate the above cells, irradiate each well for 10 min;

[0122] (4) The 96-well plate was returned to the incubator for 24 h, and then the cell viability was tested using the MTT method.

[0123] DAF ·+ The results of the photothermal killing experiments of MDA-MB-231 and HeLa by @CB[8] are as follows Fig.10 and Fig.11 If CB[8] is not added to stabilize the free radical, DAF ·+ It does not have the photothermal killing effect on cells by itself; after adding CB[8], DAF ·+ @CB[8] showed good photothermal killing effect on tumor cells. Under the above experimental conditions, DAF ·+ @CB[8] has a photothermal inhibition rate of about 80% for MDA-MB-231 and HeLa.

[0124] At the same time, Examples 2 to 8 also have a photothermal killing effect on tumor cells similar to that of the supramolecular free radicals in Example 1, indicating that the supramolecular free radicals prepared in Examples 4 to 8 also have the same biocompatibility, which will not be described in detail here.

[0125] In summary, compared with the original guest cationic radical (structural formula see Formula II), the supramolecular cationic radical has better water solubility, longer radical lifetime, stronger near-infrared absorption ability, better photothermal conversion ability and stronger photothermal killing ability of pathogens and tumor cells.

[0126] Furthermore, the supramolecular cationic free radical provided by the present invention has better water solubility, longer free radical lifetime, stronger near-infrared absorption ability, more excellent photothermal conversion ability and stronger photothermal killing ability of pathogens and tumor cells, and is expected to be developed into a photothermal therapy agent for anti-infection and tumor treatment.

[0127] In addition, the inventors of this case also referred to the aforementioned embodiments and conducted experiments with other raw materials, process operations, and process conditions described in this specification, and obtained relatively ideal results.

[0128] The various aspects, embodiments, features and examples of the present invention should be considered as illustrative in all aspects and are not intended to limit the present invention, the scope of the present invention is defined only by the claims. Other embodiments, modifications and uses will be apparent to those skilled in the art without departing from the spirit and scope of the claimed invention.

[0129] The use of headings in the present invention is not meant to limit the present invention; both the Summary and Detailed Description sections may apply to any aspect, embodiment, or feature of the present invention.

[0130] Throughout the present disclosure, where a composition is described as having, comprising, or including particular components, or where a process is described as having, comprising, or including particular process steps, it is contemplated that the compositions taught by the present invention also consist essentially of, or consist of, the recited components, and that the processes taught by the present invention also consist essentially of, or consist of, the recited process steps.

[0131] Unless specifically stated otherwise, use of the terms "include," "includes," "including," "have," "has," or "having" should generally be construed as open ended and non-limiting.

[0132] It should be understood that the order of the various steps or the order in which a particular action is performed is not very important, as long as the teachings of the present invention remain operable. In addition, two or more steps or actions may be performed simultaneously.

[0133] The above are only preferred embodiments of the present invention, which do not limit the protection scope of the present invention. For those skilled in the art, the present invention may have various modifications and changes. Within the spirit and principle of the present invention, changes, modifications, replacements, integrations and parameter changes to these embodiments by conventional substitutions or capable of achieving the same functions without departing from the principles and spirit of the present invention all fall within the protection scope of the present invention.

Claims

1. A near-infrared absorbing supramolecular cation radical, characterized in that: The structural formula of the supramolecular cation radical contains a macrocyclic molecule; The macrocyclic molecule is one or a combination of two or more of cucurbituril, cyclodextrin, calixarene and pillararene; The structural formula of the supramolecular cation radical is shown in Formula III: Wherein, X is one of methylene, carbonyl, S, O, and NH; R1 and R2 are independently any one of substituted or unsubstituted alkyl, alkyloxy, and primary / secondary / tertiary amine.

2. The near-infrared absorbing supramolecular cation radical according to claim 1, characterized in that: It can produce absorption in the range of 500 to 1100 nm.

3. A method for preparing a supramolecular cationic radical according to claim 1 or 2, characterized in that: Firstly, a supramolecular host compound and a guest molecular compound are prepared into a self-assembled complex, and then an oxidant is added to oxidize the complex to lose an electron, thereby obtaining the supramolecular cation free radical.

4. The method for preparing supramolecular cationic radicals according to claim 3, characterized in that: The supramolecular main compound is one or a combination of two or more of cucurbituril, cyclodextrin, calixarene and pillararene; And / or, the structure of the guest molecule compound is as shown in Formula I; Wherein, X is methylene, carbonyl, S, O, or NH; R1 and R2 are any one of substituted or unsubstituted alkyl, alkyloxy, or primary / secondary / tertiary amine.

5. The method for preparing supramolecular cationic radicals according to claim 3, characterized in that: The oxidant may be one or a combination of two or more of oxygen, an atmosphere containing oxygen, peroxide, ammonium persulfate, a free radical initiator, hydrogen peroxide+ferrous ion, hydrogen peroxide+catalase, ferric chloride, silver nitrate, and iodine; Preferably, the atmosphere containing oxygen is air; Preferably, the peroxide is H2O2; Preferably, the free radical initiator is 12959.

6. Use of the supramolecular cationic free radical as claimed in claim 1 or 2 in photothermal killing of pathogens.

7. The use according to claim 6, characterized in that The application includes adding supramolecular cationic radicals into an aqueous solution containing pathogenic bacteria to achieve the purpose of antibacterial; Preferably, the pathogen is a Gram-positive or Gram-negative bacterium.

8. Use of the supramolecular cationic free radical as claimed in claim 1 or 2 in photothermal killing of tumor cells.

9. The use according to claim 8, characterized in that The application comprises adding the supramolecular cationic free radical into a culture medium containing tumor cells.

10. The use according to claim 8, characterized in that The tumor cells are cervical cancer cells and breast cancer cells.

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