CN114392232B - Preparation of photoresponsive micelle for co-delivering carbon monoxide and formaldehyde molecules and application of photoresponsive micelle in synergistic DOX (Dox) anticancer - Google Patents

Preparation of photoresponsive micelle for co-delivering carbon monoxide and formaldehyde molecules and application of photoresponsive micelle in synergistic DOX (Dox) anticancer Download PDF

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CN114392232B
CN114392232B CN202210054192.9A CN202210054192A CN114392232B CN 114392232 B CN114392232 B CN 114392232B CN 202210054192 A CN202210054192 A CN 202210054192A CN 114392232 B CN114392232 B CN 114392232B
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胡进明
于露露
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Abstract

The invention discloses a preparation method of a photoresponsive micelle for co-delivering carbon monoxide and formaldehyde molecules and application of the photoresponsive micelle in synergistic DOX (Dox of X-ray fluorescence) anticancer, which comprises the steps of firstly carrying out ether reaction on a molecule containing an o-nitrobenzyl group and a 3-hydroxyflavone derivative to prepare a micromolecular donor molecule; then the donor molecule is polymerized by reversible addition-fragmentation chain transfer to obtain a polymer; and further assembling to obtain an assembly uniformly dispersed in the aqueous solution. The photoresponsive micelle prepared by the invention is stable in a water phase, can not spontaneously release carbon monoxide and formaldehyde, but can release carbon monoxide and formaldehyde molecules under the irradiation of low-intensity visible light, and after anticancer drugs such as DOX are loaded on the nano micelle, the joint release of DOX, carbon monoxide and formaldehyde can be realized, so that the synergistic effect on cancer cells can be realized.

Description

Preparation of photoresponsive micelle for co-delivering carbon monoxide and formaldehyde molecules and application of photoresponsive micelle in synergistic DOX (Dox X) anticancer
Technical Field
The invention relates to preparation of a gas signal molecule carrier and application thereof in a high-molecular medical material, in particular to preparation of a photoresponsive micelle for co-delivering carbon monoxide and formaldehyde molecules and application thereof in synergistic DOX (Dox anticancer).
Background
In the early 20 th century, CO was considered to be a colorless, odorless, toxic gas until later, researchers discovered that heme oxygenase could be expressed in organisms and this enzyme could catalyze the production of CO continuously in organisms, suggesting that CO may have important physiological roles and functions. With further research, CO is found to be a gas messenger molecule involved in regulating physiological processes of the human nervous system, immune system and cardiovascular system. Therefore, CO-releasing carbon monoxide donor molecules (CORMs) have attracted attention as potential therapeutic agents.
Endogenous CO is mainly generated by catalyzing and decomposing heme by red blood oxygenase, and exogenous CORM comprises a metal coordination compound, cyclohexamethylene tetramine carboxyl borane, a 3-hydroxyflavone derivative and the like. CORMs are classified according to the method of activating CO release, including water-triggered, light-triggered, solvent-triggered, enzyme-triggered, oxidation-triggered, pH-triggered, and the like. Most CORMs release CO in aqueous solution, but this release is uncontrolled, while light-triggered CORMs remain stable in aqueous buffers under dark conditions, releasing appropriate amounts of CO at appropriate times when exposed to light of specific wavelengths. The 3-hydroxyflavone derivative is used as a CO donor material without metal, and can controllably release CO under the irradiation of visible light (410 nm).
The o-nitrobenzyl compounds are a class of photosensitive compounds. In 1901, silber first discovered a photochemical change in the ortho-nitrobenzyl group, in the 60's of the 20 th century, barhop and Woodward used the ortho-nitrobenzyl group as a protecting group in the absence of light, and then deprotected in the presence of light. The o-nitrobenzyl group is widely applied to a light-operated drug release system as a photosensitive group, a polymer which is photosensitive and releases antibacterial gas molecules becomes a research hotspot, and the polymer of which each repeating unit in the polymer main chain contains the o-nitrobenzyl group can better realize the light-operated release of gas or drug molecules.
Formaldehyde (FA) is not only an air pollutant but also an endogenous Reactive Carbonyl Species (RCS). The research shows that endogenous formaldehyde has certain cardiovascular effect in physiological level, can damage vascular endothelial cells, is related to the incidence of atherosclerosis and can cause vasodilatation. Early studies suggested the hypothesis that endogenous formaldehyde might act as a novel gaseous signaling molecule in the body. However, only relevant studies are very limited, and there is still a great gap in the current cardiovascular effects of endogenous formaldehyde. It is worth noting that there have been several papers reporting that formaldehyde may have the effect of inducing apoptosis at home and abroad, however, experimental data are needed to prove that the mechanism of induction needs to be studied intensively.
With the development of nanotechnology, the application of biological nanomaterials provides a new opportunity for tumor treatment and shows great application prospects. Compared with the traditional medicine carrier, the nano particle medicine carrier has the characteristics of small particle size and narrow distribution. The nano-scale particle size can also enable the drug-loaded particles to show better permeability at a target position, improve the stability of the drug, play a role in slow release, and achieve the target effect by modifying the surface of the particles. Among the nanomaterials used as drug carriers, polymer nanomaterials are receiving increasing attention as a novel drug carrier. The inner core of the polymer can be used as a container of hydrophobic drugs to solubilize the drugs in the core, so that toxic and side effects are reduced, and the nano-scale particle size and the hydrophilic action of the outer shell enable the drugs not to be easily phagocytized by reticuloendothelial tissues and can prevent the adsorption of proteins and cells, so that the drugs can circulate in blood for a long time and keep stable. Most photoresponsive polymers are connected with photosensitive groups, some of the photoresponsive polymers are transported to tumor positions after being coated with drugs and irradiated with certain wavelength, the polymer structure is broken to release the drugs or change polarity, hydrophilicity and hydrophobicity and the like, and the photoresponsive polymer has certain application in the field of drug release. Sun et al report that o-nitrobenzyl ester is used for connecting hydrophilic hyaluronic acid and hydrophobic stearyl chain to synthesize a novel light-controlled nano polymer hyaluronic acid-o-nitrobenzyl-stearyl chain (HA-NB-SC), and NB HAs photolabile property and is decomposed under ultraviolet light to generate carboxyl. The diameter of the nanoparticle of the amphiphilic polymer micelle loaded with DOX is about 139nm, the micelle disintegrates under the irradiation of ultraviolet light (365 nm) to release DOX, and HeLa cell proliferation can be remarkably inhibited. Therefore, the development of a photoresponse nano-drug carrier triggered in the visible light range is an important matter of research.
Disclosure of Invention
The invention aims to provide a preparation method of a photoresponsive micelle for co-delivering carbon monoxide and formaldehyde molecules and application of the photoresponsive micelle in synergistic DOX anticancer. The nano micelle is stable in a water phase, can not spontaneously release carbon monoxide and formaldehyde, but can release carbon monoxide and formaldehyde molecules under the irradiation of low-intensity visible light. After the anticancer drug such as DOX is loaded on the nano micelle, the co-release of DOX, carbon monoxide and formaldehyde can be realized, so that the synergistic effect on cancer cells can be realized.
The invention firstly provides donor molecules with fluorescence property and capable of releasing carbon monoxide and formaldehyde in response to light, and the structure of the donor molecules is shown as the following formula (I):
Figure BDA0003475735210000021
wherein, R is 1 For responsive protection primitives, R 2 Is methyl or hydrogen, X is oxygen or Nitrogen (NH), Y is oxygen or sulfur, and Z is Nitrogen (NH) or oxygen.
The R is 1 Selected from the formula (R) shown below 1 -1) to formula (R) 1 Any one of the structures of-5):
Figure BDA0003475735210000031
preferably, X is oxygen or Nitrogen (NH) and Y is elemental oxygen or elemental sulfur.
Further, the donor molecule is preferably any one of the structures of formulae (I-1) to (I-6) shown below:
Figure BDA0003475735210000032
the invention also provides a photoresponse carbon monoxide and formaldehyde releasing donor molecular polymer with fluorescence property, which has a general structural formula shown as the following formula (II):
Figure BDA0003475735210000041
wherein, R is 1 For responsive protection primitives, R 2 Is methyl or hydrogen, X is oxygen or nitrogen, Y is oxygen or sulfur, and Z is nitrogen or oxygen. n is a radical of an alkyl radical 1 Is 20 to 70,n 2 Is 10 to 30.
The preparation method of the photoresponse micelle for co-delivering the carbon monoxide and the formaldehyde molecule comprises the steps of firstly carrying out ether reaction on a molecule containing an o-nitrobenzyl group and a 3-hydroxyflavone derivative to prepare a micromolecule donor molecule, then obtaining a polymer by reversible addition-fragmentation chain transfer polymerization (RAFT) of the donor molecule, and further assembling to obtain an assembly body which is uniformly dispersed in an aqueous solution. The method specifically comprises the following steps:
step 1: mixing the 3-hydroxy flavone derivative with potassium carbonate according to the proportion of 1: mixing at an equivalent ratio of 1, dissolving DMF, and adding 1.2 times equivalent of R 1 Stirring the compound with the structure at room temperature for reaction for 12 hours to obtain a precursor product; adding a small amount of catalyst DBTL, removing water by azeotropic distillation with toluene, taking DMF as a solvent, adding 1.8 times of equivalent of ethyl methacrylate into a reaction system, and reacting overnight at 30 ℃ to obtain a donor molecule with a structure shown in formula (I);
the structural general formula of the 3-hydroxyflavone derivative is
Figure BDA0003475735210000042
Wherein X is oxygen or Nitrogen (NH) and Y is oxygen or sulfur.
The R is 1 The compound of structure (la) is one of the compounds having the following structure:
Figure BDA0003475735210000043
Figure BDA0003475735210000044
and so on.
Step 2: reacting a compound of formula (iii) with the donor molecule obtained in step 1 in a molar ratio of 1: mixing 15 equivalent weight, adding DMSO for dissolving and clarifying, adding 0.2 times equivalent weight of initiator AIBN, and reacting for 10h in oil bath at 70 ℃ in a dark place to obtain amphiphilic carbon monoxide and formaldehyde donor molecular polymer with the structure of formula (II);
Figure BDA0003475735210000051
and step 3: and (3) self-assembling the amphiphilic carbon monoxide and formaldehyde donor molecular polymer obtained in the step (2) to obtain the photoresponse micelle nano particle. Preparing the amphiphilic carbon monoxide and formaldehyde donor molecular polymer obtained in the step 2 into a THF solution with the concentration of 2mg/mL, adjusting the stirring speed to 1000rmp, quickly injecting 1mL into 4mL deionized water, stirring for 15min, and standing for half an hour. And transferring the assembly aqueous solution into a dialysis membrane (MWCO 1.4 kDa), dialyzing by using deionized water to remove the organic solvent, and replacing the deionized water every four hours to obtain the photoresponse micelle nano particles.
According to the invention, the compound with the structure of formula (III) and the photoresponsive carbon monoxide with fluorescence property and formaldehyde donor molecule with the structure of formula (I) are mixed and reacted to obtain the amphiphilic carbon monoxide and formaldehyde donor polymer with the structure of formula (II), wherein the method of the mixed reaction is not particularly limited by the invention, and a person skilled in the art can select a suitable preparation process according to the common knowledge in the field.
The application of the light-responsive micelle for co-delivering carbon monoxide and formaldehyde molecules is to use the light-responsive micelle nano particles as a gas signal molecule carrier to load a macromolecular drug, so that the synergistic treatment of cancer cell lines can be realized.
The polymer drug comprises anticancer drugs, such as adriamycin, cyclophosphamide, daunorubicin, mitoxantrone, methotrexate, vinblastine, vindesine, etoposide, teniposide, etc.
The photoresponsive carbon monoxide and formaldehyde donor molecule with the structure shown in the formula (I) can be uniformly and stably dispersed in a water phase, can controllably release carbon monoxide and formaldehyde under the illumination of 410nm, has good targeting property, space-time controllability and biocompatibility compared with the existing gaseous signal molecule donor, and has better treatment effect compared with a single anticancer medicament after the polymer nano particles obtained by the application are loaded with DOX.
The invention provides a preparation method of a photoresponse carbon monoxide and formaldehyde donor with fluorescence property and a derivative thereof, and the photoresponse carbon monoxide and formaldehyde donor molecule with fluorescence property shown in formula (I) can realize the controlled release of carbon monoxide and formaldehyde under the irradiation of visible light by selecting a proper structure and a proper method. The invention provides a novel nano-carrier with stable and controllable release of gaseous molecules, which can jointly deliver two gaseous molecules, and changes can be monitored by fluorescence during the release process.
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FIG. 1 shows the NMR spectrum (a), the carbon spectrum (b), the mass spectrum (c) and the HPLC spectrum (d) of carbon monoxide and formaldehyde donor small molecules obtained in example 1 of the present invention.
FIG. 2 is a mass spectrum analysis of the degradation mechanism of carbon monoxide and formaldehyde donor small molecules obtained in example 1 of the present invention under 410nm illumination.
FIG. 3 shows a polymer PEG provided by an embodiment of the present invention 45 -b-PCOFA 15 The hydrogen spectrum (a) and the gel permeation chromatography (b) of (1).
Fig. 4 is a TEM photograph of nanoparticles prepared from the polymer provided in the example of the present invention.
FIG. 5 is a graph showing the fluorescence change curves (a), 453nm and 493nm (b) and 607nm (c) of nanoparticles prepared from the polymer according to the embodiment of the present invention.
FIG. 6 shows PEG obtained in this example 45 -b-PCOFA 15 And (3) analysis curves of carbon monoxide release under pure water condition of the polymer micelle nano particles and under 410nm illumination in phosphate buffer.
FIG. 7 shows PEG obtained in example 3 45 -b-PCOFA 15 The formaldehyde release under 410nm light was monitored with a formaldehyde kit for an analytical curve and a standard curve established from the absorbance at 622 nm. Wherein (a) is the ultraviolet absorption spectrum of 0.1g/L assembly under the illumination of 410nm along with the change of time, and (b) is the absorption intensity change curve at 622 nm; (c) The ultraviolet absorption spectra obtained by monitoring with the kit for formaldehyde aqueous solutions of different concentrations are shown in (d) which is a standard curve based on the absorbance at 622nm in (c).
FIG. 8 shows the synergistic effect of the polymer nanoparticles loaded (a) and unloaded (b) DOX drug obtained in this example 3 on human cervical cancer cells (HeLa) after 20 minutes of illumination at 410 nm.
FIG. 9 shows the nuclear magnetic hydrogen spectrum (a), the carbon spectrum (b), the mass spectrum (c) and the polymer PEG of the carbon monoxide micromolecules obtained in example 4 of the invention 45 -b-PCORM 13 Hydrogen spectrum (d) of (a).
Detailed Description
In the following, the technical solutions of the embodiments of the present invention will be described in detail, clearly and completely, and it is obvious that the described cases are only a small part of the embodiments. All other embodiments, which can be obtained by a person skilled in the art without making any creative effort based on the embodiments in the present invention, belong to the protection scope of the present invention.
Example 1: preparation of photoresponsive carbon monoxide and formaldehyde donor molecule CORM (wherein R is an o-nitrobenzyl group with two methoxy groups, X is an oxygen element, and Y is an oxygen element) with fluorescent properties.
Figure BDA0003475735210000061
The preparation method comprises the following steps: weighing 1 (2g, 10.74mmol), adding into 20mL ethanol to form a suspension, adding sodium hydroxide (10.8mL, 10M, 54mmol), stirring at room temperature for 30min, adding p-hydroxymethylbenzaldehyde (1.5g, 111mmol) into a reaction bottle, stirring at room temperature for 5h, cooling to 0 ℃ after the reaction is finished, dropwise adding hydrogen peroxide (4.0mL, 30%) into the reaction bottle, and stirring overnight; acidifying with 0.5M HCl to pH 6.5 to form yellow precipitate, vacuum filtering, washing with ethanol, and drying in vacuum drying oven to obtain product; weighing the product (576 mg, 2mmol) obtained in the previous step and potassium carbonate (280mg, 2mmol), adding 20mL DMF for dissolving for later use, weighing reactant 2 (626 mg,2.4 mmol), adding into a reaction bottle, stirring at room temperature for 12h after mixing, precipitating into water after reaction, performing suction filtration, fully washing the filter cake with water, and performing suction drying to obtain the product; weighing the product (300mg, 0.55mmol) in the last step, adding 3 drops of DBTL, adding 15mL of toluene for azeotropic dehydration for three times, adding 30mL of DMF as a solvent after the dehydration is finished, adding ethyl methacrylate (150mg, 0.99mmol) into a bottle, reacting at 30 ℃ overnight, spin-drying the solvent after the reaction is finished, and washing with ether for multiple times to obtain the product monomer CORM.
The obtained structure of the photoresponse carbon monoxide and formaldehyde donor molecule CORM is subjected to the characterization of nuclear magnetic hydrogen spectrum, carbon spectrum, mass spectrum and high performance liquid chromatography, and the result is shown in figure 1. As can be seen from fig. 1, the structure of the synthesized monomer was the same as expected.
The degradation mechanism of the obtained monomer CORM is researched, the result is shown in figure 2, and figure 2 is mass spectrum analysis of the degradation mechanism of the monomer CORM obtained in example 1 of the invention under the illumination of 410 nm. As can be seen from FIG. 2, the degradation mechanism of CORM is that the o-nitrobenzyl group falls off under the illumination of 410nm to form a flavone structure with exposed hydroxyl and release formaldehyde, so that the fluorescence is enhanced, and under the continuous illumination, ring opening is carried out to release CO, so that a fluorescent-free substance is formed.
Example 2: light-responsive carbon monoxide and formaldehyde release donor PEG 45 -b-PCROM 15 Synthesis of (2)
Figure BDA0003475735210000071
The PEG macroRAFT agent and the donor molecule which can release carbon monoxide and formaldehyde with photoresponse are subjected to reversible addition-fragmentation chain transfer polymerization (RAFT) to obtain the hydrophilic polymer material. The degree of polymerization n of the hydrophobic segment can be varied by varying the polymerization parameters and polymerization conditions, preferably n =10 to 30, and the degree of polymerization n is not critical to the invention, as long as the invention has no detrimental effect. To help understanding more clearly, the polymer PEG with n =15 was chosen below 45 -b-PCORM 15 The description is given for the sake of example.
The preparation method comprises the following steps: 70mg of monomer (0.1 mmol), 15mg of PEO-based macroRAFT agent ((0.0067 mmol) and 0.22mg of AIBN (0.00134 mmol) were added to the block tube, dissolved in 0.14ml of DMSO, the block tube was degassed, stirred at 70 ℃ for 10h, dissolved in tetrahydrofuran during the work-up, precipitated to zeroFiltering in water and diethyl ether, and vacuum drying overnight to obtain light yellow polymer PEG 45 -b-PCORM 15 。
The structure of the polymer obtained is characterized by nuclear magnetic hydrogen spectrum and gel permeation chromatography, and the result is shown in figure 3, and it can be seen from the figure that the synthesized polymer of the invention has the same expected structure.
Example 3: preparation of nanomaterials
Adding 1mg PEG 45 -b-PCORM 15 Dissolving in 1mL tetrahydrofuran, rapidly adding 4mL rapidly stirred water, stirring stably for 30min, placing the solution into a dialysis membrane after stirring, dialyzing with plasma water, and dialyzing for 12h to remove organic solvent to obtain the assembly. The obtained assembly was subjected to TEM characterization, and as shown in fig. 4, it was shown that the assembly was a nanoparticle having a particle size of about 21nm.
Example 4: preparing donor molecules which have fluorescent properties and release carbon monoxide independently in response to light (wherein R is an o-nitrobenzyl group with two methoxy groups, X is an oxygen element, and Y is an oxygen element).
Figure BDA0003475735210000081
The synthesis method is similar to that of example 1, the obtained photoresponsive carbon monoxide donor small molecules are subjected to the characterization of nuclear magnetic hydrogen spectrum, carbon spectrum and mass spectrum, and the same method as that of example 2 is adopted, and the PEG macroRAFT agent and the donor molecules which only release carbon monoxide in photoresponse are subjected to reversible addition-fragmentation chain transfer polymerization (RAFT) to obtain the hydrophilic polymer material. The results of the characterization of the small molecule monomers and polymers are shown in FIG. 9. As can be seen from fig. 9, the structural analysis of the synthesized monomers and polymers was the same as expected.
Application example 1: change in fluorescence properties of assemblies under light conditions
The assembly was illuminated with a 410nm LED lamp and the change in spectroscopic properties of the assembly under illumination was determined. Specifically, 1mL of the assembly is placed in a fluorescence cuvette and irradiated by 410nm, and the change of the fluorescence property of the assembly is observed along with the change of the illumination time.
As can be seen from FIG. 5, as the illumination time increases, the initial fluorescence increases, the o-nitrobenzyl group falls off under illumination and releases formaldehyde, exposing the hydroxyl group, and ESIPT occurs, resulting in strong red fluorescence. And releasing carbon monoxide after continuous illumination, and eliminating ESIPT effect, so that the fluorescence is obviously reduced, and finally the red fluorescence disappears.
Application example 2: carbon monoxide release by illumination
The quantitative detection of carbon monoxide is carried out under 410nm illumination by using a An Paer APEG-J gas detector, specifically, 10mL of assembly solution is taken in a 20mL sample bottle, and the change of the carbon monoxide content is tracked under 410nm illumination.
The results are shown in fig. 6, and fig. 6 is a detection spectrum of the assembly releasing carbon monoxide under illumination by using a An Paer APEG-J gas detector, and it can be seen from the graph that the reading of the machine cat gradually increases with the increase of illumination time, and finally reaches 19ppm.
Application example 3: releasing formaldehyde by illumination
The micelle nanoparticles were illuminated with a 410nm LED lamp, the illuminated dispersion was mixed with a formaldehyde kit according to the instructions, briefly, 3-methyl-2-benzothiazolinone hydrazone hydrochloride hydrate was added to the illuminated assembly dispersion in equal volume, and the mixture was incubated at 30 ℃ for 20 minutes in the dark. Then adding an equal volume of ferric sulfate solution, continuing co-culturing for 10 minutes, and quantifying the release amount of formaldehyde according to a standard curve established by absorbance at 622 nm. The results are shown in FIG. 7. FIG. 7 shows that the absorbance of the assembly at 622nm was monitored by UV spectrophotometer under illumination, and it can be seen that the absorbance gradually increased with the increase of illumination time, which proves that the formaldehyde emission gradually increased, and the final emission reached 30 μ M.
Application example 4: loading and synergistic anticancer of self-assembled nano particle DOX
Human cervical cancer cells (HeLa) were seeded in 96-well plates at an initial density of 10000 cells/well and 100. Mu.L of DMEM complete medium was added. After overnight culture, heLa cells were negativeAfter co-culturing assemblies (0.0125, 0.025,0.05, 0.1mg/mL) loaded/unloaded with DOX at various concentrations for 4 hours, assemblies that were not endocytosed were washed out with PBS, and one group was treated with 410nm (28 mW/cm) 2 ) The LED lights are illuminated for 20min, and the other group is not illuminated and only washed. Incubation was then continued for 24h, after which 10 μ LMTT solution was added per well, further incubation for 4h, then the suspension was aspirated off and DMSO was added to 96 well plates (100 μ L/well). The absorbance at 490nm was recorded with a microplate reader, and the cell viability was determined.
The results are shown in fig. 8, fig. 8 is an inhibition effect of DOX-loaded/unloaded assemblies obtained in example 4 on HeLa cells. From fig. 8, it can be seen that the micelle not loaded with DOX has a certain inhibition effect on the growth of HeLa cells after releasing carbon monoxide and formaldehyde in photoresponse, and the cell survival rate is reduced to about 80% under the condition that the micelle concentration is 0.1g/L and the illumination is carried out for 20min at 410 nm; under the same condition, the micelle loaded with DOX has the calculated Drug Loading Capacity (DLC) and Drug Loading Efficiency (DLE) of 8.2 percent and 18 percent respectively, and the survival rate of cells is reduced to 20 percent.
The above description of the embodiments is only intended to facilitate the understanding of the method of the invention and its core idea. It should be noted that, for those skilled in the art, it is possible to make modifications and changes to the present invention without departing from the principle of the present invention, and those modifications and changes are also within the scope of the present invention as defined in the appended claims.

Claims (6)

1. A donor molecule for the photoresponsive release of carbon monoxide and formaldehyde with fluorescent properties, characterized in that it has the following structure:
Figure FDA0003835606400000011
2. a photoresponsive carbon monoxide and formaldehyde releasing donor molecular polymer with fluorescence properties is characterized in that the polymer has a structural general formula shown as the following formula (II):
Figure FDA0003835606400000012
3. the preparation method of the light-responsive micelle for co-delivering carbon monoxide and formaldehyde molecules is characterized in that:
firstly, preparing a donor molecule shown in the formula (I) in claim 1 by carrying out ether reaction on a molecule containing an o-nitrobenzyl group and a 3-hydroxyflavone derivative; then polymerizing the donor molecule by reversible addition-fragmentation chain transfer to obtain a polymer of formula (II) according to claim 2; and further assembling to obtain an assembly uniformly dispersed in the aqueous solution.
4. The method according to claim 3, characterized by comprising the steps of:
step 1: mixing the 3-hydroxy flavone derivative with potassium carbonate according to the proportion of 1: mixing at an equivalent ratio of 1, dissolving DMF, and adding 1.2 times of R 1 Stirring the compound with the structure at room temperature for reaction for 12 hours to obtain a precursor product; adding a catalyst DBTL, removing water by azeotropic distillation with toluene, taking DMF as a solvent, adding 1.8 times of equivalent of ethyl methacrylate into a reaction system, and reacting at 30 ℃ to obtain a donor molecule with a structure shown in a formula (I);
the structural formula of the 3-hydroxyflavone derivative is shown in the specification
Figure FDA0003835606400000013
Wherein X is oxygen and Y is oxygen;
the R is 1 A compound of structure (la) is:
Figure FDA0003835606400000021
step 2: reacting a compound of formula (iii) with the donor molecule obtained in step 1 in a molar ratio of 1: mixing 15 equivalent weight, adding DMSO, dissolving, clarifying, adding 0.2 times equivalent weight of initiator AIBN, and reacting in oil bath at 70 deg.C in dark place for 10 hr to obtain amphiphilic carbon monoxide and formaldehyde donor molecular polymer with structure of formula (II);
Figure FDA0003835606400000022
and step 3: and (3) self-assembling the amphiphilic carbon monoxide and formaldehyde donor molecular polymer obtained in the step (2) to obtain the photoresponse micelle nano particle.
5. Use of the photo-responsive micelle for co-delivering carbon monoxide and formaldehyde molecules prepared by the preparation method according to claim 3 or 4, wherein: the light-responsive micelle nano particle is used for preparing a gas signal molecule carrier, and the gas signal molecule carrier is used for loading a high-molecular drug.
6. Use according to claim 5, characterized in that:
the polymer drug comprises anticancer drug selected from adriamycin, cyclophosphamide, daunorubicin, mitoxantrone, methotrexate, vinblastine, vindigoxin, etoposide or teniposide.
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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110437150A (en) * 2019-06-06 2019-11-12 中国科学技术大学 Carbon monoxide donor molecule with photoluminescent property and its preparation method and application
CN112661740A (en) * 2020-12-29 2021-04-16 中国科学技术大学 Donor molecule for photoresponse cooperative release of carbon monoxide and nitric oxide, derivative thereof, preparation method and application

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110437150A (en) * 2019-06-06 2019-11-12 中国科学技术大学 Carbon monoxide donor molecule with photoluminescent property and its preparation method and application
CN112661740A (en) * 2020-12-29 2021-04-16 中国科学技术大学 Donor molecule for photoresponse cooperative release of carbon monoxide and nitric oxide, derivative thereof, preparation method and application

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